research article epidemiology of visceral leishmaniasis in...

Hindawi Publishing CorporationBioMed Research InternationalVolume 2013 Article ID 405083 6 pageshttpdxdoiorg1011552013405083

Research ArticleEpidemiology of Visceral Leishmaniasis in a Reemerging Focusof Intense Transmission in Minas Gerais State Brazil

Ricardo Andrade Barata1 Jennifer Cunha Peixoto1 Aline Tanure1

Marcela Esteves Gomes1 Estefacircnia Conceiccedilatildeo Apolinaacuterio1 Emerson Cotta Bodevan1

Holbiano Saraiva de Arauacutejo2 Edelberto Santos Dias3 and Aimara da Costa Pinheiro4

1 Universidade Federal dos Vales dos Jequitinhonha e Mucuri Diamantina MG Brazil2 HSA Gestao e Projeto Ambiental Ltda Belo Horizonte MG Brazil3 Centro de Pesquisas Rene RachouFIOCRUZ Belo Horizonte MG Brazil4 Secretaria Municipal de Saude Governador Valadares MG Brazil

Correspondence should be addressed to Ricardo Andrade Barata ricbaratahotmailcom

Received 15 April 2013 Revised 17 June 2013 Accepted 15 July 2013

Academic Editor Milton Ozorio Moraes

Copyright copy 2013 Ricardo Andrade Barata et alThis is an open access article distributed under the Creative Commons AttributionLicense which permits unrestricted use distribution and reproduction in anymedium provided the originalwork is properly cited

This study was developed in the urban area of Governador Valadares a reemerging focus of intense transmission of visceralleishmaniasis (VL) in Brazil presenting 86 human cases of VL from 2008 to 2011 The disease prevailed in males (732) withmost patients between 0 and 9 years (441) and a lethality rate of 162 A canine survey was carried out on 16529 domestic dogsin 35 districts in the area and it showed that 302 of them (4992 dogs) were positive for VL by serum assays Prevalence ratios forcanine VL varied between 136 and 534The clinical exam of 343 seropositive dogs showed that 499 of them were consideredsymptomatic with larger prevalence of canine VL being in short-furred animals (90) The entomological survey was performedin eight districts where 2539 phlebotomines were captured preferentially in the peridomicile (845) Lutzomyia longipalpis wasthe predominant species (90) suggesting its participation in the VL transmission in the area The correlation between canineprevalence and L longipalpis density was evaluated

1 Introduction

In Brazil visceral leishmaniasis (VL) or kala-azar is oneof the biggest concerns of public health due to its highmorbid-mortality in nontreated cases [1] The etiologicalagent Leishmania infantum chagasi is transmitted mainlythrough the bite of Lutzomyia longipalpis (Diptera Psycho-didae Phlebotominae) which is infected when ingestingintracellular parasites from the peripheral blood of a dog(Canis familiaris) considered not only the main domesticreservoir but also an essential link for the maintenance of theVL epidemiological chain in urban areas [2ndash4]

In the last decades the environmental modificationscaused by man the deforestation the disordered city growththe concomitant presence of L longipalpis and domesticanimals not to mention the precarious habitation conditionsof the population all contributed to the urbanization and

geographical expansion of VL in Brazil and the emergenceof new focuses or reactivation of old ones [5]

In Governador Valadares located in Rio Doce ValeyMG cases of VL were registered in the 60s [6] Whenthe visceral leishmaniasis control program was adopted themunicipality was considered as belonging to a ldquocontrolledendemicrdquo area However in the beginning of the 90s theprogram was interrupted and epidemiological surveillancehas not been carried out regularly in the region since thenwithout notification of human cases of VL up to 2007

Since 2008 human cases of VL began to be registeredin the municipality of Governador Valadares Given thissituation the present study undertakes an epidemiologicalinvestigation taking into account human and canine casesand also the phlebotomine fauna in a reemergent focus ofintense VL transmission in Brazil

2 BioMed Research International

2 Materials and Methods

21 Study Area The municipality of Governador Valadares(18∘5110158401210158401015840 Sndash41∘5610158404210158401015840W) is located in the eastern region ofMinasGerais state covering part of the RioDoce basin in thesoutheastern area of Brazil The city has 263689 inhabitantsdistributed in 150 districts [7] The region is subjected toa hot and humid climate in which temperatures vary littlethroughout the year (annual temperature average of 256∘C)Its topography is characterized by a hilly reliefThe vegetationis between the ecosystems of seasonal semidecidual forest andsavannah but due to logging in the past some native specieswere replaced by pastures with some representatives in areasof environmental protection in the municipal surroundingareas

22 Human Cases of VL Data concerning the number ofhuman cases ofVL in the urban area ofGovernadorValadaresfrom 2008 to 2011 was obtained from the EpidemiologyManagementDVSSMS that considered the municipality asan area of intenseVL transmission (average of cases in the last5 years ge 44 cases) in the state of Minas Gerais [5]

23 Canine VL Survey The canine VL (CVL) survey wascarried out from 2008 to 2011 in all dogs domiciled inneighborhoods in the urban area with records of humancases in Governador Valadares Blood samples were obtainedannually in all the districts through the cervical or jugularvein puncture for serum production A first screening forthe presence of anti-Leishmania immunoglobulin in canineserum was performed by ELISA (Bio-ManguinhosFiocruzRJ Brazil) [8] The diagnosis confirmation was accomplishedthrough the indirect immunofluorescence assay (IFAT) [9]in agreement with the procedure adopted by the BrazilianMinistry of Health [5] Among the positive animals for IFATa random sample of 343 dogs was analyzed according to theclinical characteristics of the animal They were organizedinto asymptomatic or symptomatic groups according to theabsence or presence of at least one sign of VL infection(ie cutaneous lesions onychogryphosis keratoconjunctivi-tis weight loss emaciation and rigidity of subsequent limbs)[10] In addition they were classified in short-furred (ieBasset Beagle Boxer Brazilian Fila Doberman PinscherPit-bull Rottweiler Shar pei Weimaraner and Mongrel)and long-furred dogs (ie Cocker Siberian husky LabradorLhasa-apso German Shepherd and Poodle)

24 Entomological Captures Phlebotomine captures wereperformed in 8 neighborhoods in the municipality of Gover-nador Valadares (Altinopolis Mae de Deus Nossa Senhoradas Gracas Santa Helena Ilha de Araujos Santos DumontSao Raimundo and Turmalina) The neighborhoods werechosen taking into account the prevalence rates canine (highand low) Thirty-two HP light traps [11] were distributed inthe intra- and peridomicile exposed in two residences ofeach neighborhoodThe residences were chosen according tofavorable ecological conditions for their development such asthe presence of trees domestic animals and organic matter

Table 1 Distribution of human cases of VL according to age sexand lethality in Governador Valadares from 2008 to 2011

Age group(years)

VL human cases Total Lethality (119873)Female Male

0ndash9 15 23 38 441 510ndash19 3 4 7 82 120ndash29 1 3 4 47 130ndash39 1 7 8 93 140ndash49 1 16 17 198 250ndash59 0 6 6 69 160ndash69 2 3 5 58 2gt70 0 1 1 12 1Total 23 63 86 100 14

Sand flies were captured with traps assembled at 400 pmand removed the following day at 800 am for 3 consecutivenights each month between May 2011 and January 2012 Thespecimens captured were conditioned into hemolysis tubescontaining alcohol (70) and in accordance with Young andDuncan [12]

25 Statistical Analysis The Shapiro-Wilk test was used toevaluate the normality of data distribution (Software R v2141) The Spearman correlation between the prevalence ofcanine infection and the density of L longipalpis was calcu-lated and its statistical significance assessed (119875 value le 005)

26 Ethical Aspects This study was conducted in accor-dance with the ethical principles of animal experimentationadopted by the Brazilian College of Animal Experimentation(COBEA) and it was approved by the Ethical Committeeson the Use of Animals of Universidade Federal dos Vales doJequitinhonha eMucuri (CEUAUFVJM01010 DiamantinaBrazil)

3 Results

The municipality of Governador Valadares presented 86autochthonous cases of visceral leishmaniasis from 2008 to2011 with 14 cases in 2008 27 in 2009 24 in 2010 and 21in 2011 (data not showed) The disease prevailed in males(732) in children aged 0ndash9 years (441) with a lethalityrate of 162 (Table 1)

The canine survey carried out in 35 districts of Gover-nador Valadares from 2008 to 2011 resulted in 4992 (302)VL-seropositive dogs among 16529 tested (Table 2) CanineVL was widespread throughout the urban area with preva-lence rates varying from 136 (Sao Paulo district) to 534(Carapina district) (Table 2) Among the 343 seropositive dogsamples 499 were considered symptomatic with a largerprevalence of CVL in short-furred dogs (90) (Figure 1(a))The most frequent clinical signs in symptomatic animalswere located ulcers and onycohgryphosis as evidenced inFigure 1(b)

BioMed Research International 3

Table 2 Median prevalence of canine visceral leishmaniasis infec-tion and number of human cases by district in GovernadorValadares from 2008 to 2011

DistrictsNumber of

dogsexamined

IFAT-positivedogs

Prevalence()

VL humancases

Altinopolis 2237 756 338 10Atalaia 47 18 383 1Carapina 189 101 534 2Centro 605 121 200 6Esperanca 274 104 379 2Esplanada 166 85 512 1Fraternidade 262 63 240 1Gra-Duquesa 891 332 372 2Ilha dosAraujos 690 127 184 2

JK 259 58 224 1Jardim doTrevo 683 185 271 3

Lourdes 824 327 397 6Maria Eugenia 205 69 336 1Mae de Deus 401 132 329 3MonteCarmelo NU NU mdash 1

N Sra dasGracas 725 275 379 4

Palmeiras 764 238 311 7Planalto 354 124 350 2Santa Efigenia 195 54 277 1Santa Helena 1536 581 378 7SantaTerezinha 342 90 263 1

Santo Antonio 519 148 285 3SantosDumont 256 35 137 1

Sao Cristovao 268 64 239 2Sao Jose 165 24 145 1Sao Paulo 785 107 136 1Sao Raimundo 25 8 320 1Turmalina 946 194 205 3Vila Bretas 528 116 219 4Vila Imperio 226 46 203 1Vila Isa 230 68 295 1Vila Mariana 460 218 474 1Vila dosMontes 19 6 316 1

Vila Ozana 161 48 298 1Vila Rica 292 70 240 1Total 16529 4992 302 86NU not undertaken

The phlebotomine fauna in Governador Valadares urbanarea consisted of 4 species Lutzomyia cortelezzii (Brethes

Table 3 Phlebotomine sandflies captured with HP trap in Gover-nador Valadares by species sex and environment (from May 2011to January 2012)

SpeciesEnvironment

Total Inside OutsideC D C D

Lutzomyia cortelezzii 26 13 79 63 181 71L intermedia 3 2 3 10 18 07L longipalpis 80 250 221 1733 2284 900L whitmani 1 2 0 1 4 01Lutzomyia spp 10 6 11 25 52 21Subtotal 120 273 314 1832 2539 100

1923) Lutzomyia intermedia (Lutz and Neiva 1912) Lut-zomyia longipalpis (Lutz and Neiva 1912) and Lutzomyiawhitmani (Antunes andCoutinho 1939) totaling 2539 speci-mens ofwhich 2105weremales (83) and434 females (17)The peridomicile presented the largest percentage of thecaptured specimens (845) The predominant species wasL longipalpis (90) (Table 3)The phlebotomine distributionaccording to the neighborhoods andmonths of collection canbe visualized in Table 4

The correlation between the density of the L longipalpiscaptured and canine prevalence is represented in Figure 2There was no statistically significant association betweenthe variables number of sandflies and prevalence of canineinfection (119875 = 0057)

4 Discussion

In Brazil VL is considered a neglected disease that prevails inplaces where underprivileged social conditions predominateIt contributes to the maintenance of the inequality picturethat we have nowadays as it represents a strong barrier to thesocial and economical development of the country [5] In thelast decades the urbanization phenomenon has been pointedout as the responsible for the appearance of new focuses andreemergence of old ones in urban areas of small andmedium-sized cities [13ndash15]

Governador Valadares located in the Southeastern areaof Brazil is an example of a reemerging focus of VL as aresult of urbanization The VL cases notified occurred wherehousing conditions basic sanitation and garbage collectionwere poor The residents have low socioeconomic statusliving together with domestic animals and accumulatedorganic matter (data not shown) The same conditions wereevidenced in other places where VL transmission is endemic[16 17]

Analyzing the human cases according to the age group itwas noticed that VL was more frequent in children under 10years old (441) (Table 1) The same findings were reportedby several authors [18ndash20] The lethality rate was 162Nascimento et al [14] and Queiroz et al [21] verified alethality rate of 115 and 102 respectively The greatestprevalence of VL in children (Table 1) can explain the highlethality rate found as they are yet incompletely developedimmune system


Table 4 Phlebotomine sandflies captured with HP trap in Governador Valadares by district and sex (fromMay 2011 to January 2012)

Year Months

Districts

119873 Altinopolis Mae de N Sra Santa Ilha dos Santos Sao TurmalinaDeus Gracas Helena Araujos Dumont Raimundo

C D C D C D C D C D C D C D C D

2011

May 17 28 58 278 8 27 32 98 0 0 0 1 0 0 0 1 548 216Jul 8 34 20 153 7 49 19 72 0 0 0 0 0 2 0 0 364 144Sep 3 5 4 8 5 10 7 29 0 0 1 5 0 0 0 0 77 30Nov 21 33 15 7 8 25 11 57 3 1 0 0 0 1 3 4 189 74

2012 Jan 19 299 99 315 56 549 10 14 0 0 0 0 0 0 0 0 1361 536Total 467 957 744 349 4 7 3 8 2539 100

0 100 200 300

Long

Short

CVL (N)

Fur t

ype

SymptomaticAsymptomatic

(a)

0 50 100 150

Apathy

Alopecia

Ectoparasites

Emaciation

Generalised ulcers

Keratoconjunctivitis

Localized ulcers

Onycogryphosis

Paresia on the feet

Frequency (N)

Clin

ical

sign

s CV

L

(b)

Figure 1 Frequencies of canine visceral leishmaniasis by fur type and clinical status (a) and the recorded clinical signs of CVL (b) in urbandogs of Governador Valadares MG Brazil

The prevalence of canine-VL in Brazil has been demon-strated to be between 19 and 35 in endemic areas [22ndash26]In a previous study undertaken in Governador ValadaresMalaquias et al [27] evidenced 137 of positivity in dogs inurban areas In the present study the average prevalence was302 but in some neighborhood it reached 534The highprevalence of canine infection has been given as one of therisk factors for VL occurrence [28 29]

In this context symptomatic or asymptomatic seroposi-tive dogs play an important role in the maintenance of theinfection [30] Signs of VL were detected in 171 (499) ofthe 343 dogs sampled The pattern of clinical signs observedfor the disease most commonly included weight loss apathyand emaciation similar to what was observed by Silva et al[15] The most worrying fact is the percentage of dogs withnonapparent infection (501) as they show a high degree ofcutaneous parasitism being capable of living together withthe parasite for long periods [31 32]

In addition short-furred dogs represented the largestpercentage of seropositive animals (90) suggesting thatshort fur determines larger chance by a greater propensity tocontract the infection in the population investigated because

it enables the phlebotomine access to the place of the bite[33 34] These results are reinforced by Franca-Silva et al[4] who observed a larger prevalence of CVL in short-furreddogs

In Minas Gerais several authors have showed the abun-dance of L longipalpis in urban areas where VL is endemic[16 35 36] The same pattern has been observed in otherBrazilian areas [37 38] where the species clearly participatesin the transmission of L infantum chagasi However inprevious research in the same area as the present studyalthough the authors found L longipalpis it was not the mostabundant species This fact may be because the area sampledwas a transitional one betweenwild and urban environmentsThus the predominance of this sand fly in an urban area withhigh frequencies in residences and surroundings suggestsits participation in the transmission of L infantum chagasiamong dogs and humans in the city of Governador Valadares[39]

The presence of a great number of seropositive dogs andthe high density of L longipalpis have been affirmed to bethe main risk factors for the occurrence of VL in urban areas[40 41] In the present study the correlation between these


0

2

4

6

8

10

12

0

5

10

15

20

25

30

35

40A

ltinoacute

polis

Matildee

de D

eus

N S

ra d

as G

raccedila

s

Sant

a Hele

na

Ilha d

os A

rauacutej

os

Sant

os D

umon

t

Satildeo

Raim

undo

Turm

alin

a

DistrictsVL human casesCanine prevalence ()L longipalpis ()

Llongipalpismdash

Cani

ne p

reva

lenc

e (

)

VL

hum

an ca

ses (

2008

ndash201

1)

Figure 2 Distribution of Lutzomyia longipalpis population preva-lence of canine infection and human cases VL per district ofGovernador Valadares MG Brazil

two variables presented no statistical significance Howeverif we analyze Figure 2 we notice that there is a tendency foran increase in canine prevalence in neighborhoods where thedensity of L longipalpis is higher such as Altinopolis NossaSenhora das Gracas Mae de Deus and Santa Helena

Thus these factors seem to be decisive for the occurrenceof VL in the city of Governador Valadares reinforcing theneed for rigid controlling actions through the euthanasiaof seropositive dogs the use of residual insecticide andenvironmental management in residences as well as rigorousepidemiological surveillance

Acknowledgments

The authors would like to thank Fundacao de Amparo aPesquisa do Estado de Minas Gerais (APQ-009810) for thefinancial support

References

[1] C M F Gontijo and M N Melo ldquoLeishmaniose visceral noBrasil quadro atual desafios e perspectivasrdquo Revista Brasileirade Epidemiologia vol 7 no 3 pp 338ndash349 2004

[2] L M Deane and M P Deane ldquoVisceral leishmaniasis in Brazilgeographical distribution and trnsmissionrdquo Revista do Institutode Medicina Tropical de Sao Paulo vol 4 pp 198ndash212 1962

[3] R J Quinnell O Courtenay L Garcez and C Dye ldquoThe epide-miology of canine leishmaniasis transmission rates estimatedfrom a cohort study in Amazonian Brazilrdquo Parasitology vol 115no 2 pp 143ndash156 1997

[4] J C Franca-Silva R T da Costa A M Siqueira et al ldquoEpi-demiology of canine visceral leishmaniosis in the endemic area

of Montes Claros Municipality Minas Gerais State BrazilrdquoVeterinary Parasitology vol 111 no 2-3 pp 161ndash173 2003

[5] Brasil Manual de vigilancia e controle da leishmaniose visceralMS Serie A Normas e Manuais Tecnicos Ministerio da SaudeSecretaria de Vigilancia em Saude Departamento de VigilanciaEpidemiologica Brasılia Brazil 2006

[6] M V Coelho and A R Falcao ldquoAspectos epidemiologicos docalazar emMinas Geraisrdquo Jornal Brasileiro de Medicina vol 10no 1 pp 259ndash262 1966

[7] IBGE ldquoAnuario Estatıstico do BrasilmdashInstituto Brasileiro deGeografia e Estatısticardquo 2010

[8] A Voller W H Bidwell and I Arjona The Enzyme LinkedImmunossorbent Assay (ELISA) A Guide with Abstracts ofMicroplate Applications Dynatec Europe 1979

[9] M E Camargo and C Rebonato ldquoCross-reactivity in fluores-cence tests for Trypanosoma and Leishmania antibodies A sim-ple inhibition procedure to ensure specific resultsrdquo AmericanJournal of Tropical Medicine and Hygiene vol 18 no 4 pp 500ndash505 1969

[10] FManciantiMGramiccia L Gradoni and S Pieri ldquoStudies oncanine leishmaniasis control 1 Evolution of infection of differ-ent clinical forms of canine leishmaniasis following antimonialtreatmentrdquoTransactions of the Royal Society of TropicalMedicineand Hygiene vol 82 no 4 pp 566ndash567 1988

[11] H Pugedo R A Barata J C Franca-Silva et al ldquoHP ummodelo aprimorado de armadilha luminosa de succao para acaptura de pequenos insetosrdquo Revista da Sociedade Brasileira deMedicina Tropical vol 38 no 2 pp 70ndash72 2005

[12] D G Young and M A Duncan ldquoGuide to the identificationand geographic distribution of Lutzomyia sand flies in Mexicothe West Indies Central and South America (Diptera Psycho-didae)rdquo Memoirs of the American Entomological Institute vol54 no 1 pp 1ndash881 1994

[13] C H N Costa H F Pereira and M V Araujo ldquoVisceralleishmaniasis epidemic in the State of the Piauı Brazil (1980ndash1986)rdquoRevista de Saude Publica vol 24 no 5 pp 361ndash372 1990

[14] M D S B Nascimento J M L Costa B I P Fiori et alldquoAspectos epidemiologicos determinantes na manutencao daleishmaniose visceral no Estado do Maranhao Brasilrdquo Revistada Sociedade Brasileira de Medicina Tropical vol 29 no 3 pp233ndash240 1996

[15] E S Silva C M F Gontijo R S Pacheco V O P Fiuza andR P Brazil ldquoVisceral leishmaniasis in the Metropolitan Regionof Belo Horizonte State of Minas Gerais Brazilrdquo Memorias doInstituto Oswaldo Cruz vol 96 no 3 pp 285ndash291 2001

[16] R A Barata J C F D Silva R T D Costa et al ldquoPhlebotominesand flies in Porteirinha an area of American visceral leish-maniasis transmission in the State of Minas Gerais BrazilrdquoMemorias do Instituto Oswaldo Cruz vol 99 no 5 pp 481ndash4872004

[17] E M Michalsky J C Franca-Silva R T Costa et al ldquoLeish-maniose Visceral estudo de flebotomıneos e infeccao caninaem Montes Claros Minas Gerais Brasilrdquo Revista da SociedadeBrasileira de Medicina Tropical vol 38 no 2 pp 147ndash152 2005

[18] R Badaro T C Jones R Lorenco et al ldquoA prospective study ofvisceral leishmaniasis in an endemic area of BrazilrdquoThe Journalof Infectious Diseases vol 154 no 4 pp 639ndash649 1986

[19] M C A Marzochi K B F Marzochi and R W CarvalholdquoVisceral leishmaniasis in Rio de Janeirordquo Parasitology Todayvol 10 no 1 pp 37ndash40 1994


[20] J M L Costa G M C Viana A C R Saldanha et al ldquoLeish-maniose visceral no Estado do Maranhao Brasil Evolucao deuma epidemiardquo Cadernos de Saude Publica vol 11 no 2 pp321ndash324 1995

[21] M J A Queiroz J G B Alves B Jailson et al ldquoLeishmaniosevisceral caracterısticas clınico-epidemiologicas em criancas dearea endemicardquo Jornal de Pediatria vol 80 no 2 pp 141ndash1462004

[22] I A Sherlock and S P Almeida ldquoNotas sobre leishmaniosecanina no Estado da Bahiardquo Revista Brasileira de Malariologiae Doencas Tropicais vol 22 no 2 pp 231ndash242 1970

[23] L B Iversson M E Camargo A Villanova et al ldquoInqueritosorologico para pesquisa de leishmaniose visceral empopulacao canina-urbana do municıpio de Sao Paulo-Brasil(1979ndash1982)rdquo Revista do Instituto de Medicina Tropical de SaoPaulo vol 25 no 6 pp 310ndash317 1983

[24] S G Coutinho M P Nunes M C Marzochi and N Tra-montano ldquoA survey for American cutaneous and visceralleishmaniasis among 1342 dogs from areas in Rio de Janeiro(Brazil) where the human diseases occurrdquoMemorias do InstitutoOswaldo Cruz vol 80 no 1 pp 17ndash22 1985

[25] T G Evans I A B Vasconcelos J W Lima et al ldquoCaninevisceral leishmaniasis in northeast Brazil assessment of serodi-agnostic methodsrdquo American Journal of Tropical Medicine andHygiene vol 42 no 2 pp 118ndash123 1990

[26] M P Nunes J M Jackson R W Carvalho N J Furtado andS G Coutinho ldquoSerological survey for canine cutaneous andvisceral leishmaniasis in areas at risk for transmission in Riode Janeiro where prophylactic measures had been adoptedrdquoMemorias do Instituto Oswaldo Cruz vol 86 no 4 pp 411ndash4171991

[27] L C CMalaquias R do Carmo Romualdo J B do Anjos Jr RC Giunchetti R Correa-Oliveira and A B Reis ldquoSerologicalscreening confirms the re-emergence of canine leishmaniosisin urban and rural areas in Governador Valadares Vale do RioDoce Minas Gerais Brazilrdquo Parasitology Research vol 100 no2 pp 233ndash239 2007

[28] J B F Vieira and G E Coelho ldquoLeishmaniose visceral oucalazar aspectos epidemiologicos e de controlerdquo Revista daSociedade Brasileira de Medicina Tropical vol 31 supplement2 pp 85ndash92 1998

[29] L C P de Oliveira R R de Araujo C R Alves E Mouta-Confort J A Lopez and FW deMendonca-Lima ldquoSeropreva-lence and risk factors for canine visceral leishmaniasis in theendemic area of Dias DrsquoAvila State of Bahia Brazilrdquo Revista daSociedade Brasileira deMedicinaTropical vol 43 no 4 pp 400ndash404 2010

[30] E M Michalsky M F Rocha A C V M da Rocha Lima etal ldquoInfectivity of seropositive dogs showing different clinicalforms of leishmaniasis to Lutzomyia longipalpis phlebotominesand fliesrdquo Veterinary Parasitology vol 147 no 1-2 pp 67ndash762007

[31] MC AMarzochi P C Sabroza LM Toledo et al ldquoLeishman-iose Visceral na cidade do Rio de JaneiromdashBrasilrdquo Cadernos deSaude Publica vol 1 no 1 pp 5ndash17 1985

[32] M D F Madeira A D O Schubach T M P Schubach C ALeal and M C D A Marzochi ldquoIdentification of Leishmania(Leishmania) chagasi isolated fromhealthy skin of symptomaticand asymptomatic dogs seropositive for leishmaniasis in themunicipality of Rio de Janeiro Brazilrdquo The Brazilian Journal ofInfectious Diseases vol 8 no 6 pp 440ndash444 2004

[33] EDMoreira Jr VMM de SouzaM SreenivasanN L LopesR B Barreto and L P de Carvalho ldquoPeridomestic risk factorsfor canine leishmaniasis in urban dwellings new findings froma prospective study in BrazilrdquoThe American Journal of TropicalMedicine and Hygiene vol 69 no 4 pp 393ndash397 2003

[34] F S Juliao B M P S Souza D S Freitas et al ldquoInvestigacao deareas de risco como metodologia complementar ao controle daleishmaniose visceral caninardquo Pesquisa Veterinaria Brasileiravol 27 no 8 pp 319ndash324 2007

[35] E M Michalsky J C Franca-Silva R A Barata et al ldquoPhle-botominae distribution in Janauba an area of transmission forvisceral leishmaniasis in BrazilrdquoMemorias do Instituto OswaldoCruz vol 104 no 1 pp 56ndash61 2009

[36] E S Dias S Regina-Silva J C Franca-Silva et al ldquoEco-epidemiology of visceral leishmaniasis in the urban area ofParacatu state of Minas Gerais BrazilrdquoVeterinary Parasitologyvol 176 no 2-3 pp 101ndash111 2011

[37] A G de Oliveira E A B Galati O de Oliveira et al ldquoAbun-dance of Lutzomyia longipalpis (Diptera Psychodidae Phle-botominae) and urban transmission of visceral leishmaniasis inCampo Grande state of Mato Grosso do Sul BrazilrdquoMemoriasdo Instituto Oswaldo Cruz vol 101 no 8 pp 869ndash874 2006

[38] NAMissawa andE SDias ldquoPhlebotomine sandflies (DipteraPsychodidae) in the municipality of Varzea Grande an areaof transmission of visceral leishmaniasis in the state of MatoGrosso Brazilrdquo Memorias do Instituto Oswaldo Cruz vol 102no 8 pp 913ndash918 2007

[39] R A Barata G F Paz M C Bastos et al ldquoPhlebotomine sand-flies (Diptera Psychodidae) in Governador Valadares a trans-mission area for American tegumentary leishmaniasis in Stateof Minas Gerais Brazilrdquo Revista da Sociedade Brasileira deMedicina Tropical vol 44 no 2 pp 136ndash139 2011

[40] C Margonari C R Freitas R C Ribeiro et al ldquoEpidemiologyof visceral leishmaniasis through spatial analysis in Belo Hori-zonte municipality state of Minas Gerais BrazilrdquoMemorias doInstituto Oswaldo Cruz vol 101 no 1 pp 31ndash38 2006

[41] R Lainson and B F Rangel ldquoLutzomyia longipalpis and theeco-epidemiology of American visceral leishmaniasis withparticular reference to Brazilmdasha reviewrdquoMemorias do InstitutoOswaldo Cruz vol 100 no 8 pp 811ndash827 2005

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2 Materials and Methods

21 Study Area The municipality of Governador Valadares(18∘5110158401210158401015840 Sndash41∘5610158404210158401015840W) is located in the eastern region ofMinasGerais state covering part of the RioDoce basin in thesoutheastern area of Brazil The city has 263689 inhabitantsdistributed in 150 districts [7] The region is subjected toa hot and humid climate in which temperatures vary littlethroughout the year (annual temperature average of 256∘C)Its topography is characterized by a hilly reliefThe vegetationis between the ecosystems of seasonal semidecidual forest andsavannah but due to logging in the past some native specieswere replaced by pastures with some representatives in areasof environmental protection in the municipal surroundingareas

22 Human Cases of VL Data concerning the number ofhuman cases ofVL in the urban area ofGovernadorValadaresfrom 2008 to 2011 was obtained from the EpidemiologyManagementDVSSMS that considered the municipality asan area of intenseVL transmission (average of cases in the last5 years ge 44 cases) in the state of Minas Gerais [5]

23 Canine VL Survey The canine VL (CVL) survey wascarried out from 2008 to 2011 in all dogs domiciled inneighborhoods in the urban area with records of humancases in Governador Valadares Blood samples were obtainedannually in all the districts through the cervical or jugularvein puncture for serum production A first screening forthe presence of anti-Leishmania immunoglobulin in canineserum was performed by ELISA (Bio-ManguinhosFiocruzRJ Brazil) [8] The diagnosis confirmation was accomplishedthrough the indirect immunofluorescence assay (IFAT) [9]in agreement with the procedure adopted by the BrazilianMinistry of Health [5] Among the positive animals for IFATa random sample of 343 dogs was analyzed according to theclinical characteristics of the animal They were organizedinto asymptomatic or symptomatic groups according to theabsence or presence of at least one sign of VL infection(ie cutaneous lesions onychogryphosis keratoconjunctivi-tis weight loss emaciation and rigidity of subsequent limbs)[10] In addition they were classified in short-furred (ieBasset Beagle Boxer Brazilian Fila Doberman PinscherPit-bull Rottweiler Shar pei Weimaraner and Mongrel)and long-furred dogs (ie Cocker Siberian husky LabradorLhasa-apso German Shepherd and Poodle)

24 Entomological Captures Phlebotomine captures wereperformed in 8 neighborhoods in the municipality of Gover-nador Valadares (Altinopolis Mae de Deus Nossa Senhoradas Gracas Santa Helena Ilha de Araujos Santos DumontSao Raimundo and Turmalina) The neighborhoods werechosen taking into account the prevalence rates canine (highand low) Thirty-two HP light traps [11] were distributed inthe intra- and peridomicile exposed in two residences ofeach neighborhoodThe residences were chosen according tofavorable ecological conditions for their development such asthe presence of trees domestic animals and organic matter

Table 1 Distribution of human cases of VL according to age sexand lethality in Governador Valadares from 2008 to 2011

Age group(years)

VL human cases Total Lethality (119873)Female Male

0ndash9 15 23 38 441 510ndash19 3 4 7 82 120ndash29 1 3 4 47 130ndash39 1 7 8 93 140ndash49 1 16 17 198 250ndash59 0 6 6 69 160ndash69 2 3 5 58 2gt70 0 1 1 12 1Total 23 63 86 100 14

Sand flies were captured with traps assembled at 400 pmand removed the following day at 800 am for 3 consecutivenights each month between May 2011 and January 2012 Thespecimens captured were conditioned into hemolysis tubescontaining alcohol (70) and in accordance with Young andDuncan [12]

25 Statistical Analysis The Shapiro-Wilk test was used toevaluate the normality of data distribution (Software R v2141) The Spearman correlation between the prevalence ofcanine infection and the density of L longipalpis was calcu-lated and its statistical significance assessed (119875 value le 005)

26 Ethical Aspects This study was conducted in accor-dance with the ethical principles of animal experimentationadopted by the Brazilian College of Animal Experimentation(COBEA) and it was approved by the Ethical Committeeson the Use of Animals of Universidade Federal dos Vales doJequitinhonha eMucuri (CEUAUFVJM01010 DiamantinaBrazil)

3 Results

The municipality of Governador Valadares presented 86autochthonous cases of visceral leishmaniasis from 2008 to2011 with 14 cases in 2008 27 in 2009 24 in 2010 and 21in 2011 (data not showed) The disease prevailed in males(732) in children aged 0ndash9 years (441) with a lethalityrate of 162 (Table 1)

The canine survey carried out in 35 districts of Gover-nador Valadares from 2008 to 2011 resulted in 4992 (302)VL-seropositive dogs among 16529 tested (Table 2) CanineVL was widespread throughout the urban area with preva-lence rates varying from 136 (Sao Paulo district) to 534(Carapina district) (Table 2) Among the 343 seropositive dogsamples 499 were considered symptomatic with a largerprevalence of CVL in short-furred dogs (90) (Figure 1(a))The most frequent clinical signs in symptomatic animalswere located ulcers and onycohgryphosis as evidenced inFigure 1(b)



DistrictsNumber of

dogsexamined

IFAT-positivedogs

Prevalence()

VL humancases











SpeciesEnvironment





4 Discussion






Year Months

Districts



2011

May 17 28 58 278 8 27 32 98 0 0 0 1 0 0 0 1 548 216Jul 8 34 20 153 7 49 19 72 0 0 0 0 0 2 0 0 364 144Sep 3 5 4 8 5 10 7 29 0 0 1 5 0 0 0 0 77 30Nov 21 33 15 7 8 25 11 57 3 1 0 0 0 1 3 4 189 74

2012 Jan 19 299 99 315 56 549 10 14 0 0 0 0 0 0 0 0 1361 536Total 467 957 744 349 4 7 3 8 2539 100

0 100 200 300

Long

Short

CVL (N)

Fur t

ype


(a)

0 50 100 150

Apathy

Alopecia

Ectoparasites

Emaciation

Generalised ulcers


Localized ulcers

Onycogryphosis

Paresia on the feet

Frequency (N)

Clin

ical

sign

s CV

L

(b)









0

2

4

6

8

10

12

0

5

10

15

20

25

30

35

40A

ltinoacute

polis

Matildee

de D

eus

N S

ra d

as G

raccedila

s

Sant

a Hele

na

Ilha d

os A

rauacutej

os

Sant

os D

umon

t

Satildeo

Raim

undo

Turm

alin

a


Llongipalpismdash

Cani

ne p

reva

lenc

e (

)

VL

hum

an ca

ses (

2008

ndash201

1)




Acknowledgments


References



















































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology



DistrictsNumber of

dogsexamined

IFAT-positivedogs

Prevalence()

VL humancases











SpeciesEnvironment





4 Discussion






Year Months

Districts



2011

May 17 28 58 278 8 27 32 98 0 0 0 1 0 0 0 1 548 216Jul 8 34 20 153 7 49 19 72 0 0 0 0 0 2 0 0 364 144Sep 3 5 4 8 5 10 7 29 0 0 1 5 0 0 0 0 77 30Nov 21 33 15 7 8 25 11 57 3 1 0 0 0 1 3 4 189 74

2012 Jan 19 299 99 315 56 549 10 14 0 0 0 0 0 0 0 0 1361 536Total 467 957 744 349 4 7 3 8 2539 100

0 100 200 300

Long

Short

CVL (N)

Fur t

ype


(a)

0 50 100 150

Apathy

Alopecia

Ectoparasites

Emaciation

Generalised ulcers


Localized ulcers

Onycogryphosis

Paresia on the feet

Frequency (N)

Clin

ical

sign

s CV

L

(b)









0

2

4

6

8

10

12

0

5

10

15

20

25

30

35

40A

ltinoacute

polis

Matildee

de D

eus

N S

ra d

as G

raccedila

s

Sant

a Hele

na

Ilha d

os A

rauacutej

os

Sant

os D

umon

t

Satildeo

Raim

undo

Turm

alin

a


Llongipalpismdash

Cani

ne p

reva

lenc

e (

)

VL

hum

an ca

ses (

2008

ndash201

1)




Acknowledgments


References



















































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology



Year Months

Districts



2011

May 17 28 58 278 8 27 32 98 0 0 0 1 0 0 0 1 548 216Jul 8 34 20 153 7 49 19 72 0 0 0 0 0 2 0 0 364 144Sep 3 5 4 8 5 10 7 29 0 0 1 5 0 0 0 0 77 30Nov 21 33 15 7 8 25 11 57 3 1 0 0 0 1 3 4 189 74

2012 Jan 19 299 99 315 56 549 10 14 0 0 0 0 0 0 0 0 1361 536Total 467 957 744 349 4 7 3 8 2539 100

0 100 200 300

Long

Short

CVL (N)

Fur t

ype


(a)

0 50 100 150

Apathy

Alopecia

Ectoparasites

Emaciation

Generalised ulcers


Localized ulcers

Onycogryphosis

Paresia on the feet

Frequency (N)

Clin

ical

sign

s CV

L

(b)









0

2

4

6

8

10

12

0

5

10

15

20

25

30

35

40A

ltinoacute

polis

Matildee

de D

eus

N S

ra d

as G

raccedila

s

Sant

a Hele

na

Ilha d

os A

rauacutej

os

Sant

os D

umon

t

Satildeo

Raim

undo

Turm

alin

a


Llongipalpismdash

Cani

ne p

reva

lenc

e (

)

VL

hum

an ca

ses (

2008

ndash201

1)




Acknowledgments


References



















































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology


0

2

4

6

8

10

12

0

5

10

15

20

25

30

35

40A

ltinoacute

polis

Matildee

de D

eus

N S

ra d

as G

raccedila

s

Sant

a Hele

na

Ilha d

os A

rauacutej

os

Sant

os D

umon

t

Satildeo

Raim

undo

Turm

alin

a


Llongipalpismdash

Cani

ne p

reva

lenc

e (

)

VL

hum

an ca

ses (

2008

ndash201

1)




Acknowledgments


References



















































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology

research article epidemiology of visceral leishmaniasis in...

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