6th INTERNATIONAL SYMPOSIUMon PHLEBOTOMINE SANDFLIES

October 27-31, 2008, Lima, PERU

40º Aniversario

6th INTERNATIONAL SYMPOSIUMon PHLEBOTOMINE SANDFLIES

October 27-31, 2008, Lima, PERU

40º Aniversario

6th INTERNATIONAL SYMPOSIUMon PHLEBOTOMINE SANDFLIES

October 27-31, 2008, Lima, PERU

40º Aniversario

6th INTERNATIONAL SYMPOSIUMon PHLEBOTOMINE SANDFLIES

October 27-31, 2008, Lima, PERU

40º Aniversario

Auditorio Hugo Lumbreras Cruz, Casa Honorio Delgado Av. Armendáriz 445, Miraflores, Lima, Peru

6th

INTERNATIONAL SYMPOSIUM ON PHLEBOTOMINE SANDFLIES

The 6th

INTERNATIONAL SYMPOSIUM ON PHLEBOTOMINE SANDFLIES

(ISOPS6) is being held in Lima, Peru on the 95th

anniversary of the description of the

first sandfly species reported for Peru, Lutzomyia verrucarum (Townsend, 1913). This

species is the vector of both bartonellosis (Carrion's disease, verruga peruana or Oroya

fever), and Andean cutaneous leishmaniosis (uta) in Peru. This year we also

commemorate the 40th

anniversary of the Instituto de Medicina Tropical Alexander von

Humboldt, founded by Prof. Hugo Lumbreras, who devoted his life to research and

training in tropical medicine .

ISOPS6 attendees have an unique opportunity to visit the historically significant places

of Peruvian bartonellosis and leishmaniosis transmission areas, including the Verrugas

bridge, and the type locality of L. verrucarum, area where many workers died during the

construction of the Central Railroad, 1870-1872; since the destination was the mining

town of La Oroya, they called the unknown disease "Oroya fever". Since 1913, sandfly

research in Peru is mainly addressed to the control of bartonelosis and leishmaniosis and

their vectors; these two diseases are endemic in large areas and affect high proportions

of the Peruvian population. Sandfly diversity in Peru is very high because of the many

different environments present in the Peruvian territory. To date 160 species (which will

be 168 after ISOPS6) have been reported for Peru; this number has resulted fom the

work of Peruvian and foreign scientists with a special dedication to sandfly taxonomy.

About 20 % of these sandfly species are endemic for Peru, thus providing additional

interest to the study of the transmission cycles of bartonellosis and leishmaniosis.

Kike

J. Enrique Perez

Instituto de Medicina Tropical Alexander von Humboldt

Universidad Peruana Cayetano Heredia

isops6@gmail.com, kike.perez@upch.pe

CHAIRMEN

J. Enrique Perez, Peru

Alejandro Llanos-Cuentas, Peru

LOCAL ORGANIZER:

INSTITUTO DE MEDICINA TROPICAL ALEXANDER VON HUMBOLDT

Organizing Committee:

J. Enrique Perez, Coordinator

Eduardo Gotuzzo H.

Alejandro Llanos-Cuentas

Jorge Arévalo

Ciro Maguiña

Humberto Guerra

Elena Ogusuku

Carmen Rado

Pablo Villaseca

Dionicia Gamboa

HONORARY COMMITTEE

N. Añez, Venezuela

R. Badaro, Brazil

R. Killick-Kendrick, UK

M. Maroli, Italia

J.A. Rioux, Francia

I. Sherlock, Brazil

SCIENTIFIC COMMITTEE

D. El Naiem, Sudan

N. Léger, France

P. Volf, Czech Republic

A. Warburg, Israel

P. Lawyer, USA

D. Feliciangeli, Venezuela

Elizabeth Rangel, Brazil

E.F. Rangel, Brazil

J. Enrique Perez, Peru

Reginaldo Brazil, Brazil

Edgar Rowton, USA

Michele Maroli, Italy

SPONSORS

ISOPS6 Organizing Committee is very grateful to:

Vicerrectorado de Investigacion (UPCH)

ROCHE

INMUNOCHEM

J & B Lab SAC.

CONTENTS

ORAL PRESENTATIONS

Session I: Taxonomy of Phlebotomine Sandflies. 01-04

Session II: Ecology, Distribution, and Population Dynamics of Sandflies. 05-14, 54

Session III Peruvian sandflies, leishmaniosis and bartonellosis. 15-20

Session IV: Ecology of leishmaniosis. 21-27

Session V: Detection of Leishmania in sandflies. Other microorganisms in sandflies.

28-32

Session VI: Leishmania-sandfly interaction. 33-38

Session VII: Sandfly saliva. 39-43

Session VIII: Sandfly surveillance. 44-47

Session IX: Sandfly control. 48-53, 54

POSTER PRESENTATIONS

POSTER SESSION I

I: Systematics and Taxonomy of Phlebotomine Sandflies

II Biology of Sandflies.

IIIA: Ecology, Distribution, and Population Dynamics of Sandflies.

POSTER SESSION II

IIIB: Ecology, Distribution, and Population Dynamics of Sandflies.

IV Molecular Methods for Detection of Leishmania in Sandflies.

V: Sandflies as Hosts of Microorganisms other than Leishmania.

VI Ecology of Leishmaniosis.

VII Sandfly Saliva.

VIII: Surveillance and Sandfly Control.

ANNEX I: Visit to the Rimac valley.

AUTHORS INDEX

ISOPS6 PROGRAM

Morning: 09:00-13:00

Lunch: 13:00-14:00

Afternoon: 14:00-18:00

Monday October 27 Morning: Registration

Lunch

Afternoon

Session I: Molecular Taxonomy of Phlebotomine Sandflies.

Chairman: L.E. Munstermann

Secretary: N. Leger

Session II: Ecology, Distribution, and Population Dynamics of Sandflies.

Chairman: R.P. Brazil

Secretary:. J.D. Andrade

Tuesday October 28 Morning

Session III Peruvian sandflies, transmission of leishmaniosis and bartonellosis.

Chairman: J.E. Perez

Secretary: P. Pachas

The Researchers. J. Enrique Perez

Epidemiology of bartonellosis in the geographical diversity of Peru. Paul Pachas

A gene network of Lutzomyia verrucarum based on cytochrome b genetic

variability. Abraham Caceres et al.

Entomological Surveillance System of the Ministry of Health, Peru: Sandfly

records. Elena Ogusuku

Uta and espundia. Alejandro Llanos-Cuentas

Verruga peruana. Ciro Maguiña

Lunch

Afternoon

Poster Session I

Session IV: Ecology of leishmaniosis.

Chairman: O.D. Salomon

Secretary: C. Ferro

Wednesday October 29 Full day: visit to the Rimac Valley.

Thursday October 30 Morning

Session V: Detection of Leishmania in sandflies. Other microorganisms in sandflies.

Chairman: E. Rangel

Secretary: D. Gamboa.

Session VI: Leishmania-sandfly interaction.

Chairman: P. Lawyer

Secretary: M.L. Vilela

Lunch

Afternoon

Poster Session II

Session VII: Sandfly saliva.

Chairman: P. Volf

Secretary: D.A. Elnaiem

Friday October 31 Morning

Session VIII: Sandfly surveillance.

Chairman: E. Ogusuku

Secretary: J.A. Swaby

Session IX: Sandfly control.

Chairman: E. Rowton

Secretary: G. Wasserberg

Lunch

Afternoon

Session X: Closing Session: ISOPS6 Conclusions.

Chairman: M. Maroli

Secretary: E. Rojas

Election of ISOPS7 venue

Clossing of ISOPS6

ORAL PRESENTATIONS

SESSION I: Taxonomy of Phlebotomine Sandflies

OP-01

MOLECULAR AND BEHAVIORAL DIFFERENTIATION AMONG

BRAZILIAN POPULATIONS OF LUTZOMYIA LONGIPALPIS (DIPTERA:

PSYCHODIDAE: PHLEBOTOMINAE).

ALEJANDRA S. ARAKI1, FELIPE M. VIGODER

1, LUIZ G. S. R. BAUZER

1,

GABRIEL E. M. FERREIRA1, NATALY A. SOUZA

2, IZENEIDE B. ARAÚJO

3,

JOHN G. C. HAMILTON4, REGINALDO P. BRAZIL

5, ALEXANDRE A.

PEIXOTO1

1Laboratório de Biologia Molecular de Insetos, IOC, Fundação Oswaldo Cruz, Rio de

Janeiro, Brazil. 2Laboratório de Transmissores de Leishmanioses, IOC, Fundação

Oswaldo Cruz, Rio de Janeiro, Brazil. 3Curso de Ciências Biológicas e Agrárias,

Universidade Estadual do Piauí, Parnaíba, Piauí, Brazil. 4Centre for Applied

Entomology and Parasitology, Institute of Science & Technology in Medicine, Keele

University, United Kingdom. 5Laboratório de Bioquímica e Fisiologia de Insetos, IOC,

Fundação Oswaldo Cruz, Rio de Janeiro, Brazil.

Lutzomyia longipalpis, the main vector of American visceral leishmaniasis, is a

species complex but until recently the existence of sibling species among Brazilian

populations was considered a controversial issue. In addition, there is still no consensus

regarding the number of species occurring in this complex. We combined molecular and

behavioral approaches to study the differentiation between Brazilian populations of L.

longipalpis. Using period, a gene that controls circadian rhythms and male courtship

songs in Drosophila, we analyzed the molecular polymorphism in a number of L.

longipalpis samples from different regions of Brazil and compared the results with our

previously published data using the same marker. We also studied the male copulation

songs and pheromones from some of these populations. The results obtained so far

suggest the existence of two main groups of populations in Brazil, one group

representing a single species with males producing Burst-type copulation songs and

cembrene-1 pheromones; and a second group that is more heterogeneous and probably

represents a number of incipient species producing different combinations of Pulse-type

songs and pheromones.

Financial Support : This work was supported by grants from the Howard Hughes

Medical Institute, CNPq, Capes, FAPERJ and FIOCRUZ.

OP-02

MOLECULAR TAXONOMY OF PHLEBOTOMUS NEGLECTUS AND P.

SYRIACUS, VECTORS OF LEISHMANIA INFANTUM

VIT DVORAKa, JAN VOTYPKA

a, VLADIMIR IVOVIC

b, MARIA ANTONIOU

c,

MICHELE MAROLI d

, BULENT ALTEN e and PETR VOLF

a

a

Department of Parasitology, Faculty of Science, Charles University, Vinicna 7, Prague

2, 128 44 Czech Republic, e-mail: icejumper@seznam.cz b

Department of Medical Parasitology, University of Belgrade, Belgrade, Serbia c

Medical School, University of Crete, Heraklion, Greece d

Department of Parasitology, Instituto Superiore di Sanita, Roma, Italy e Department of Biology,Hacettepe University, Ankara, Turkey

Within the subgenus Larroussius, taxonomic status and distribution of two species of

Phlebotomus major complex was studied. While P. neglectus is a proven vector of

Leishmania infantum as well as Phleboviruses, P. syriacus is assumed to be probable

vector on circumstatial evidence, namely, its distribution, biting habits and taxonomic

position.

Different populations of P. neglectus and P. syriacus from Italy, Croatia, Montenegro,

Albania, mainland Greece and island of Crete, Turkey, Syria, Lebanon and Israel were

studied by three different molecular biology techniques: RAPD analysis, internal

transcribed spacer 2 (ITS2) sequencing (nuclear marker) and cytochrome B sequencing

(mitochondrial marker). All three methods confirmed that P. neglectus is present in

north-central part of the Mediterranean (Italy, Balcan countries) while P. syriacus in the

eastern part. Turkey is the country through which goes the dividing line of both species

distribution. However, a detailed survey revealed a sympatric occurence of both species

in Crete. Since morphological identification of these two closely related species is

extremely difficult and intermediate morphological forms were found in many places an

attempt was made to characterise a molecular marker which would readily distinguish

them.

OP-03

GENETIC VARIABILITY AMONG POPULAIONS OF THE SAND FLY

LUTZOMYIA (NYSSOMYIA) WHITMANI S.L. ACCESSED BY MULTI-LOCUS

ENZYME ELECTROPHORESIS

SIMONE M COSTA¹, CLAUDIO RV MENESES², MARA G TAVARES3 &

ELIZABETH F RANGEL¹

¹Lab. Transmissores de Leishmanioses, Instituto Oswaldo Cruz/FIOCRUZ; ²University

of California, Davis, Department of Pathology, Microbiology and Immunology 3Dep.de

Biologia Geral, Universidade Federal de Viçosa,MG,Brasil

Lutzomyia (N.) whitmani s.l. is widely distributed in Brazil. It is incriminated as vector

of two dermotropic leishmania species: Leishmania (V.) braziliensis and Leishmania

(V.) shawi. The hypothesis that L. (N.) whitmani s.l. could represent a species complex

in Brazil is based on its biology and vectorial competence, which means the capacity of

this sand fly in transmitting two distinct leishmania species as well as being present in

different habitats. The aim of this study is to evaluate the variability and the genetic

structure of the Brazilian populations of L. (N.) whitmani s.l. collected in the

municipalities of Ilhéus (BA) (type locality), Londrina (PR), Santarém and Paragominas

(PA). MLEE was used because it detects the variation and estimates the degree of

similarity among species, either by numerical analysis of genetic relationships or by

taxonomic identification and classification. Samples were prepared following the

protocol of Salles et al. (1986). Five enzymes (GPI, MDH, PGM, Me and PGD) allowed

secure interpretation of five genetic loci. The mean observed heterozygosity (Ho) varied

from 0.087 ± 0.080 to 1.38 ± 0.98, and the mean expected heterozygosity (He) varied

from 0.254 ± 0.121 to 0.317 ± 0.144. All populations are in Hardy-Weinberg

equilibrium. Genetic structure analysis revealed a mean value of FIS higher than FST =

(0.493 > 0.046), suggesting some intra population differentiation. The analysis of the

genetic structure of the populations showed excess of homozygotes, suggesting low

genetic differentiation among them. They represent a single species with great chances

of having occurred subdivision in the past due to evolution. Additional molecular tools

will be employed in order to better understand the local structure of the Brazilian

populations of L. (N.) whitmani.

Financial Support: IOC/FIOCRUZ, PAPES III-FIOCRUZ, CNPq.

OP-04

THE PHYLOGENY OF SAND FLIES OF THE GENUS LUTZOMYIA

(DIPTERA: PSYCHODIDAE)

LORENZA BEATI1, SARAH MOCK

1, LYNN A. JONES

2, AND LEONARD E.

MUNSTERMANN2

1 Georgia Southern University, Statesboro, Georgia, USA

2 Yale University School of Public Health, New Haven, Connecticut, USA

Introduction. The occurrence of cryptic species, the scarcity of informative

morphological characters, and the presence of substantial intraspecific polymorphism

have rendered very complex the study of the systematic relationships among the 400

species of the phlebotomine genus Lutzomyia. Young & Duncan treated the genus as 15

unranked subgenera and 11 ‘species groups’ based on a small number of morphological

characters. These characters often have proven difficult to evaluate for identifications

due to intrataxon variation and for phylogenetic inference due to lack of information

about ancestral status.

Objectives. Mitochondrial and nuclear DNA sequence data were used to indicate the

taxonomic relationships among the taxa established in 1994 by Young & Duncan within

the Lutzomyia genus, as well as to test the strength of the more recent reorganization

proposed by Galati based on morphometric analyses.

Methods. Four gene sequences (12S rDNA, cytochrome oxidase I mtDNA, 18S rDNA,

and 28S rDNA) were obtained from 86 species representing 22 of the 26 Lutzomyia

subgenera and groups. The 4 molecular data sets, taken separately or in a total evidence

analysis, were used to infer species relationships at several taxonomic hierarchical

levels.

Results. Phylogenetic formation provided by the four data sets. Degree of congruence

among the phylogenetic trees varied with sequence and taxon compared. Clades

showing consistent composition throughout the analyses were compared to traditional

morphological grouping of species to establish which aspects of the phenotypic

taxonomy were corroborated (or disproved) by molecular analysis. In general, Galati’s

taxonomic organization based on the biometrics of morphological characters was

supported by the molecular phylogenies.

Conclusions. The 18s rDNA sequenc is conservatively evolving and supported the

basal divisions at the basal (subtribe) level. 12s and 28s rDNAs resolved several of

groups established in the Young & Duncan and the Galati schemes, in addition to

associating previously unattached species. The analyses also brought to evidence

several taxa in need of a thorough systematic re-evaluation.

Financial support. United States National Institutes of Health grants U19 AI065866,

R01 AI056254.

SESSION II: Ecology, Distribution, and Population Dynamics of Sandflies

OP-05

PHLEBOTOMINE SAND FLIES (DIPTERA: PSYCHODIDAE) OF THE

SOUTHEASTERN REGION, BRAZIL: A CURRENT LIST OF VALID SPECIES

ANDREY JOSÉ DE ANDRADE1, FILIPE DANTAS-TORRES

2 & EUNICE

APARECIDA BIANCHI GALATI3

1Departamento de Parasitologia, Universidade Federal de Minas Gerais, Belo

Horizonte, MG, Brasil - bioandrey@gmail.com.br 2Departamento de Imunologia, Centro de Pesquisas Aggeu Magalhães, Fundação

Oswaldo Cruz, Recife, PE, Brasil - fdt@cpqam.fiocruz.br 3Departamento de Epidemiologia, Faculdade de Saúde Pública, São Paulo, SP, Brasil -

egalati@usp.br

Introduction: Brazil is divided into five geographical regions: North, Northeast,

Southeast, South, and Center-West. The Southeast region is composed of four states:

Espírito Santo, Minas Gerais, Rio de Janeiro, and São Paulo. The phlebotomine sand fly

fauna of these states is well known, but no updated checklist has been published since

2003. Objective: To provide an updated list of valid species of phlebotomine sand flies

found in Southeast Brazil. Methods: The description of the distribution of

phlebotomine sand flies in Southeast Brazil is based on data obtained through a review

of the literature and is also based on the authors’ observations. Phlebotominae

taxonomy follows the classification recently proposed by Galati. Results and

Conclusions: A total of 115 phlebotomine sand fly species belonging to four subtribes

and 16 genera have been found in Southeast Brazil: BRUMPTOMYIINA: Brumptomyia

(14 spp.); LUTZOMYIINA: Evandromyia (18), Expapillata (2), Lutzomyia (12),

Migonemyia (4), Pintomyia (9), Pressatia (3), Sciopemyia (2) and Trichopygomyia (1);

PSYCHODOPYGINA: Bichromomyia (1), Martinsmyia (5), Nyssomyia (5),

Psathyromyia (13) and Psychodopygus (9) and SERGENTOMYIINA: Deanemyia (2)

and Micropygomyia (15). The state with the greatest number of recorded species is

Minas Gerais (94 spp.), followed by São Paulo (67), Rio de Janeiro (58), and Epírito

Santo (46). Approximately 50% of the phlebotomine species found in Brazil occur in

the Southeaster region. This diversity is at least partly a result of the large number of

studies carried out in this region. It also reflects the various characteristics of its diverse

biomes (e.g., Atlantic rainforest, various types of savannah and speleological provinces)

favorable to a range of different species, many of which are also well adapted to highly

urbanized areas.

OP-06

PHLEBOTOMINE (DIPTERA: PSYCHODIDAE) SANDFLLIES COLLECTED

IN A CAVE IN PALMAS, TOCANTINS STATE, BRAZIL.

JOSÉ DILERMANDO ANDRADE FILHO1, TÂMARA OLIVEIRA MACHADO

2,

LUSY DISNEY GOMES DE ANDRADE ALMEIDA3, GUSTAVO MAYR DE

LIMA CARVALHO1, CRISTIANI DE CASTILHO SANGUINETTE

1,

REGINALDO PEÇANHA BRAZIL4

1- Lababoratório de leishmanioses - Centro de Pesquisas René Rachou - Fiocruz

2-Setor de Controle das Leishmanioses, Malária e Raiva- Secretaria Municipal de Saúde

CCZ - Palmas - TO

3 -Laboratório de Entomologia - Secretaria Municipal de Saúde- CCZ - Palmas - TO

4- Laboratório de bioquímica e fisiologia de insetos - Instituto Oswaldo Cruz

Sandflies are important in public health due to ability to transmit certain pathogens,

especially leishmaniases. This work describe the sand fly species collected in a cave

located in Boa Esperança Farm (10º22.921’ S and 48º03.587’ W, 504 m.a.s.l.) in the

Taquaruçu district, about 70km from the urban center of Palmas, capital of the Brazilian

State of Tocantins. Captures were performed with HP light trap close to the entrance, in

the aphotic zone of the cave and in one house, far from about two kilometers from the

cave. The sand flies were maintained in 70% alcohol solution and taken to the “Centro

de Referência Nacional e Internacional para Flebotomíneos” (CRNIF) of the Instituto

René Rachou/Fiocruz for slide preparation; females were mounted in Berlese liquid and

males in Canada balsam. All the sand flies were deposited in the collection of CRNIF

and a total of 16 species of sandflies were identified: Evandromyia lenti (40,0%),

Edentomyia sp. (11,7%), Evandromyia carmelinoi (10,3%), Evandromyia termitophila

(9,6%), Deanemyia samueli (9,6%), Lutzomyia longipalpis (9,0%), Micropygomyia

vonatzingeni (2,1%), Sciopemyia sordellii (1,4%), Lutzomyia dispar (1,4%),

Micropygomyia longipennis (0,7%), Nyssomyia whitmani (0,7%), Micropygomyia

echinatopharynx (0,7%), Psathyromyia brasiliensis (0,7%), Micropygomyia peresi

(0,7%), Evandromyia sallesi (0,7%) and Martinsmyia oliveirai (0,7%). All 16 species

were collected in the cave, and five (include L. longipalpis) was found in the house.

Researches conducted specifically to study the sandflies in caves are rare. Our results

are very interesting as regards the diversity and density of sandflies in caves, which may

be equal to or higher than in the forests or others environments. This is the first record

of the genus Edentomyia outside the State of Piauí, Northeast Region of Brazil. Only

one species of this genus is described (i.e. E. piauiensis) and new studies about the

species found in Tocantins is necessary in order to verify their real taxonomic identity.

OP-07

BIOGEOGRAPHY OF THE ORIENTAL PHLEBOTOMINE SAND

FLIES (DIPTERA: PSYCHODIDAE)

K. ILANGO

Zoological Survey of India, Southern Regional Station, 130 Santhme High Road,

Chennai- 600 028, INDIA. E-mail: kilangozsi@rediffmail.com

The historical biogeography of Phlebotomine sand fly taxa Hertigia, Warileya,

Idiophlebotomus, Spelaeophlebotomus, Anaphlebotomus, and Euphlebotomus and the

Phlebotomus (Euphlebotomus) argentipes species complex was investigated using

phylogenetic inference from comparative genital morphology, distribution of ancestral

taxa, fossil evidence, and geological age. Idiophlebotomus and Euphlebotomus occur in

the Oriental region with one species from the northeast Australia, whereas

Anaphlebotomus occurs both in the Afrotropical and Oriental Regions. These disjunct

distribution patterns across the Oriental Region and the present day distribution are

likely to be vicariance due to break of Gondwana. Fossil records, extant taxa

distribution, phylogenetic analysis of the Old-world Phlebotominae and paleogeography

reconstructed by Metcalfe (1998) suggest that ancestors of Idiophlebotomus and

Euphlebotomus originated apparently in the Cimmerian continent of northern margin of

Gondwanaland in the early Permian (290 million years ago, MYA) and subsequently

radiated in the Mesozoic by tectonic vicariance. The Phlebotomus argentipes species

complex occurs in the South-, and South-east Asian countries, and its phylogeography

was caused probably through both vicariance and dispersal events from 5O MYA (the

Eocene) until the Recent (<10,000 y).

OP-08

CHANGE IN THE COMPOSITION AND DISTRIBUTION OF THE SPECIES

OF Lutzomyia (DIPTERA: PSYCHODIDAE) IN TEN YEARS IN THE

CENTRAL WESTERN REGION OF COLOMBIA

LINA MARIA CARRILLO1, 2, KARINA MONDRAGÓN SHEM

1, LUZ ADRIANA

ACOSTA CARDONA1, RAFAEL VIVERO

1, DANIELA VERGARA

1, ANDRÉS

VÉLEZ1, IVÁN DARÍO VÉLEZ

1

1Programa de Estudio y Control de Enfermedades Tropicales -PECET-, Universidad de

Antioquia, Medellín, Colombia 2Facultad de Ciencias Agrarias de la Universidad de Antioquia, Medellín, Colombia

Introduction: Worldwide changes such as global warming, the construction of water

dams and the destruction of forests exert their influence on the distribution and

abundance of sandflies, vectors of leishmaniasis, which in turn mark the geographical

impact of the disease. This work sought to determine the geographical and altitudinal

distribution of the species of sandflies and compare this with a similar study carried out

ten years ago. Materials and Methods: The area of study is located in the eastern slope

of the Andean Range, comprising four life zones (Holdridge, 1977), and covering an

area of approximately 870Km2. Sandflies were collected using an entomological

transect during the dry and rainy seasons of the year with adhesive, CDC and Shannon

traps. Results: 14 species were collected in this survey; almost all of them in the intra

and peridomestic habitat. Six of the species are of medical importance, and four are new

species for the area of study. Most of the species captured presented both altitude and

geographical restriction, with only four of them in more than one life zone. No

statistically meaningful differences were found between the captures in the rainy and

dry seasons. Discussion: Climate change and habitat degradation during the ten year

period have negatively affected the diversity and abundance of the sandflies, but it is the

vector species that are highly domiciliated that proved more stability and adaptation

which contributes to the endemicity of the disease. Lu. longipalpis presented a wider

altitudinal range with a new world record at 1387masl, increasing with this the areas of

risk for visceral leishmaniasis; Lu. trinidadensis was the most widely distributed species

in the area of study. Maps were created with ArcGis9.2 illustrating areas of risk for

cutaneous and visceral leishmaniasis as well as the geographical distribution of the

sandflies.

OP-09

ECOLOGY, DISTRIBUTION, AND POPULATION DYNAMICS OF SAND

FLIES IN A NEW L. TROPICA FOCUS IN THE JUDEAN DESERT

LAOR ORSHAN1, SHIMON BITTON

2, DAVID SZEKELY

3

1

Laboratory of Entomology, Ministry of Health, Jerusalem, Israel. 2 Regional Municipality Mate Binyamin, Psagot, Mizrah Binyamin, Israel.

3 Ministry of Environmental Protection, Jerusalem, Israel.

In 2004 a Leishmania tropica (LT) outbreak occurred for the first time in the town of

Ma'ale Adummim, 10 km east of Jerusalem. This outbreak extended the disease

distribution to the western margins of the Judean Desert. Sand flies are abundant in

Ma'ale Adummim, as well as in other Israeli villages situated in the Judean Desert, and

have been known as a source of nuisance for many years. In 2005 we started a

comprehensive study of phlebotomine sand flies, as part of the combined governmental

effort to prevent leishmaniasis and to understand the causes and dynamics of LT

infections. We collected over 230,000 specimens from 1373 CO2-baited trap nights in

outdoor collections. All males (44%) and random samples of females were identified to

species. Phlebotomus sergenti, the known L. tropica vector in the region, comprised

~90% of the total catch; Ph. papatasi and Ph. syriacus were collected in equal numbers

(~5% each), and Ph. tobbi was rare (0.13%). On eastern and southern slopes catches

regularly consisted of 100-200 specimens per trap. On western and northern slopes

catches were much smaller, consisting of 10-20 specimens per trap. Furthermore, small

numbers of sand flies were collected in the winter months and only in traps placed near

rock crevices. Populations increased in April, rose further in May, peaked in August and

decreased drastically in November; numbers in December were similar to those of

April.

Although Ph. sergenti is by far the most numerous species outdoors, Ph. papatasi is

dominant indoors (93%, 1150 of 1233 specimens collected by mouth aspirator in 500

visits to houses). Catches of Ph. papatasi indoors peaked in May and June, stabilized in

July, decreased significantly in November and disappeared in January, reappearing in

small numbers in April. Very few Ph. sergenti specimens were found indoors, of which

55% were collected in October and November.

The relevance of the differential spatial and temporal distribution of Ph. sergenti to the

epidemiology of the disease is discussed.

OP-10

FIRST ENTOMOLOGICAL SURVEY ON PHLEBOTOMINE SANDFLIES IN

CATANIA, ITALY, SINCE THE HISTORICAL ADLER & THEODOR’S

INVESTIGATIONS ON MEDITERRANEAN KALA AZAR (1931)

V. D’URSO1, O. LISI

1, S. DISTEFANO

1, G. BARRESI

1, M. MAROLI

2

1Department of Animal Biology “M. La Greca”, University of Catania, Italy;

2MIPI

Department, Istituto Superiore di Sanità, Rome, Italy. E-mail: dursove@unict.it

Catania, located along the Jonian coast of Mediterranean sea, is the second biggest

city of Sicily. At the beginning of the years 1930s, Shaul Adler and Oscar Theodor, on

behalf of the ‘Kala Azar Commission’ of the Royal Society, performed the historical

“Investigations on Mediterranean Kala Azar”. In these studies, the city of Catania was

referred as to one of the largest visceral leishmaniasis foci found in Sicily, with 150-200

cases recorded annually. The authors give also detailed information on the bionomics

and vector competence of the local sandfly fauna (Adler & Theodor, 1931). All local

species were found at high densities at the city periphery in proximity of animal shelters

and gardens, but they were very rare in the city centre where no suitable breeding places

were available. Particularly, while P. papatasi was present throughout the whole

sandfly distribution area, P. perniciosus was irregularly distributed. For instance, in the

same street one house could be found be infested by P. perniciosus whereas another was

entirely free, but all houses in the sandfly belt contained P. papatasi. Since the pioneer

studies of Adler and Theodor (1931), no investigations have been carried out so far in

the city of Catania.

In the present note we report the data of an entomology survey performed during

the hot season 2006 aimed to study the current distribution of sandfly fauna in the urban

environments of Catania. A total of 51 collecting sites were identified in the 44 sub-

units (600 x 600 m each) of the area under study. Three monthly collections were

carried out by using sticky traps set in the wall holes along roads. 88.2% of sites

monitored were positive for phlebotomines. The collections accounted for 4341 sandfly

specimens. Five species were indentified being Sergentomyia minuta the prevalent

species (59.6%). Among the species of the Phlebotomus genus, P. perniciosus was the

most abundant (36.5%), followed by P. sergenti (2.5%) and P. neglectus (1.5%). The

other two species were P. perfiliewi (2 specimens) and P. mascittii (1 specimens).

P. perniciosus, one of the most competent Leishmania infantum vectors in the

Mediterranean foci, was present in all the sandfly-positive sites and showed significant

densities in sites located in the city centre. The findings are discussed in relation to the

risk of Leishmania transmission in the urban area of Catania and compared with

previous Adler & Theodor’s observations.

OP-11

SANDFLIES (DIPTERA: PSYCHODIDAE) CAPTURED IN THE

SPELEOLOGICAL PROVINCE OF BAMBUÍ, DISTRICT OF MEDIUM AND

HIGH SÃO FRANCISCO, MINAS GERAIS, BRAZIL

GUSTAVO MAYR DE LIMA CARVALHO1, CRISTIANI DE CASTILHO

SANGUINETTE1, REGINALDO PEÇANHA BRAZIL

2, JOSÉ DILERMANDO

ANDRADE FILHO1

1- Laboratório de Leishmanioses - Centro de Pesquisas René Rachou - Fiocruz

2- Laboratório de Bioquímica e Fisiologia de Insetos - Instituto Oswaldo Cruz - Fiocruz

Phlebotomines are invertebrate hosts of species of the genus Leishmania and

responsible for leishmaniases in humans and other mammals. Beside leishmaniases they

are vectors of arboviruses, bartonelosis and can also cause allergic reaction due to their

bites. Caves are ecotones well differentiated of the external environment and among the

insects that live or visit the inner area and adjacent environment, sandflies are found

constantly. Several caves, even before any studies, are opened for visiting and do not

have any scientific follow up with loss of geological and biological information. The

main aim of this work was to study the sand fly fauna in a cave environment of a

Speleological Province of Bambuí, Municipality of Lassance, State of Minas Gerais.

Preliminary captures were performed with HP light traps and sandflies were mounted in

Berlese (females) and Canada Balsam (males). A total of 242 specimens of sandflies of

13 species were captured with the prevalence of Lutzomyia cavernicola (30%),

Lutzomyia sp. (25%) and Lu. sordellii (23%). We call the attention of the second species

that may be a new species of the Cortelezzii complex and will be described later. All the

other species represent 22% of the total and some are incriminated as important vectors

of leishmaniases, Lu. longipalpis, Lu. whitmani and Lu. intermedia.

Financial Support: CPqRR/FIOCRUZ, FAPERJ, FAPEMIG

OP-12

THE ECOLOGY OF CUTANEOUS LEISHMANIASIS IN SOUTHERN

ISRAEL: DEMOGRAPHIC AND SPATIAL PATTERNS IN THE VECTOR –

RESERVOIR HOST ASSOCIATION

GIDEON WASSERBERG1

1Walter Reed Army Institute of Research, Silver Spring, MD.

Understanding of the ecology of the wildlife reservoir – disease vector interaction is

essential for the control of vector-borne zoonotic diseases. In southern Israel cutaneous

leishmaniasis is caused by Leishmania major, transmitted by Phlebeotomus papatasi,

and maintained by the sand rat Psammomys obesus. In this study, I focused on the

demographic and spatial aspects of this interaction. With respect to the demographic

aspect, using a 1.5 years mark-release-recapture study, I studied the temporal dynamics

and distribution of the disease within the host. Most of the transmission occurred

following sand fly activity peak of May. Prevalence increases with age but does not

differ between sexes. Survival rate is affected by infection and gender: non-infected

females have higher survival rate than non-infected males but vice-versa with respect to

infected animals. The probability for an individual host to survive long enough to

constitute a potential infection source was estimated as 8.2%. With respect to the spatial

aspect, I manipulated the degree of burrow isolation by placing artificial burrows at

various distances from active host burrows and monitored the rates of their re-

colonization by dispersing sand flies. Artificial burrow colonization rates were highest

at 0 and 60 meters but even the farthest burrows at 120 and 240 m were frequently

colonized. I also conducted a large-scale survey of sand rat burrow distribution after

which I trapped and removed sand rats from selected burrows and monitored burrow re-

colonization after three months. I used logistic regression to analyze of the relations

between the densities of neighboring active host burrow on infection occurrence per

host at various spatial scales. Only at the scale of 500-m radius from host burrow, I

found significant positive association indicating that this is a relevant scale for

transmission. A risk calibration model, derived from the equation of the logistic model,

suggests that even complete host eradication will not nullify transmission risk thus

questioning the benefit of local host eradication strategy. The majority of colonizers

were juvenile rodents. Burrow re-colonization is dictated by the phenological state of

the Chenopodiaceae plants neighboring the burrow. Results indicate that sand flies,

more than sand-rats, are responsible for the spatial dynamics of the disease.

OP-13

MADAGASCAR : AN ELDORADO FOR PHLEBOTOMOLOGISTS.

N. LÉGER AND J. DEPAQUIT.

Only one ( or two ? ) species ( Grassomyia ) of phlebotomine sand flies has been

reported in Madagascar before we described one ( 1978) , then latter (2002-2008), 8

new species for Science from this island. Only one of them can be included without

problems in an already known sub-genus. For the others we have to suggest the

creation of new sub-genus . In addition 5, or more, other enigmatic taxons are waiting

to be described.

Parallel to morphological study, molecular biology has been used to establish males-

females status, to group species in new sub-genus ( or genus?) and to try to understand

this strange and unique fauna.

We constructed phenograms and cladograms. Our phylogenetic hypothesis have been

compared with the present knowledge of the paleogeography of Madagascar.

OP-14

THE SANDFLY FAUNA AND POPULATION SEASONALITY IN LA GRANJA

(CAJAMARCA, PERU)

RUBIN DE CELIS, MAURICIO, GUSTAVO ARBIZU & J. ENRIQUE. PEREZ

Insecticidas y Rodenticidas de Perú SAC.

The sandfly fauna of La Granja (Department of Cajamarca, Peru) was recorded monthly

from August 2007 to July 2008. Seven collection sites were selected: La Granja, La

Iraca, Checos, Paraguay, Sauce, Uñiga and Las Limas, at altitudes from 1900 to 2100

meters above sea level. Sandflies were collected using Shannon traps with protected

human bite and CDC light traps inside and outside houses, all traps were operated from

18:00 to 06:00.

15 sandfly species were identified for La Granja area, these are: Lutzomyia

maranonensis (83 %), L. castanea, L. cajamarcaensis, L. cortelezzii, L. evansi, L.

ayacuchensis, L. pallidithorax, L. pia “like”, L. pisuquia, L. robusta, L. reclusa, L.

sallesi, L. verrucarun, L. sp., and Warileya lumbrerasi.

The peak of the sandfly population was recorded in June 2008. In the study area the end

of the rainy season was not drastic as it is in the Center of Peru (the end of March), the

rainy season in the study area finished on early June, and the outbreak of the sandfly

population occurred at the end of June. This fact is due to the type of vegetation in the

area: Forest, and because of the nearest to the Equadorian Line, which makes the

environment more tropical, in despite of the altitude. The sandfly populations were very

low on the rest of the year. Sandflies were more abundant inside houses; this fact is

response to the adverse weather conditions outside (rain, wind and clouds).

This study is part of a baseline environmental study. We also thank the participation of

personnel from DISA Chota of the Ministry of Health.

SESSION III: Peruvian sandflies, leishmaniosis and bartonellosis

OP-15

THE RESEARCHERS

J. ENRIQUE PEREZ

Instituto de Medicina Tropical Alexander von Humboldt, Universidad Peruana

Cayetano Heredia. kike@upch.edu.pe

Four steps can be clearly recognized in the sandfly study in Peru, these steps correspond

with the researchers in charged of the studies. 1. 1913-1915. The scientific research on

sandflies started with C H T Townsend in 1913, who was contracted by the Peruvian

government to perform studies that concluded with the discovery of the vector of

“verruga peruana” Phlebotomus verrucarum. 2. 1925-1945. With H Noguchi, R

Shannon, M. Hertig, G B Fairchild and G Gorbitz, they did taxonomical, biological

studies and Hertig, Fairchild and Gorbitz also worked on the sandfly control with DDT,

successful studies which are still reference in the sandfly control. The Peruvian sandfly

fauna was 5-6 species. 3. 1960-1980. B Llanos, F Blancas, H Herrer, A Viana M, H

Frahia. The Peruvian sandfly fauna was around 35 species. From here, local scientists

are more interested to work on sandflies.

4. 1980 to present. D G Young, P Villaseca, C Davies, A Llanos-Cuentas, M Cameron

A Caceres, E Ogusuku, J E Perez, E Galati, R Fernandez, A Tejada, L Laughlin N

Solorzano, P Pachas and others. Studies were addressed to know epidemiological

aspects of the diseases transmission; helped with molecular tools, the detection of

pathogens in sandflies and saliva studies were posible; phylogenetical studies are also

carried out by DNA analyses. Today the Peruvian sandfly fauna is near to 170 species.

OP-16

EPIDEMIOLOGY OF BARTONELLOSIS IN THE GEOGRAPHICAL

DIVERSITY OF PERU.

PAUL PACHAS, MD

Direccion General de Epidemiologia, Ministerio de Salud, Lima, Peru. (General

Directorate of Epidemiology, Ministry of Health-Peru).

Introduction

Carrion's disease is caused by the bacterium Bartonella bacilliformis (Bb) and

transmitted by different species of Lutzomyia; classically has been described in Andean

valleys of Peru, Ecuador and Colombia. But in the past 15 years, has expanded into new

areas of transmission and has increased the number of cases, for which it is considered

as a re-emerging disease in Peru. Our goal is to describe the epidemiology of Carrion's

disease in Peru.

Methods: We review the database of the epidemiology surveillance system of the General

Directorate of Epidemiology of the Ministry of Health, from 1995 to 2008. We

considered as a confirmed case, if they had a positive blood smear, a blood culture with

growth of colonies compatible with Bb, and as an eruptive case if they had eruptive

lesions in skin or mucous membranes of red or subcutaneous nodules with less than six

months

Results:

Between 1995 to September 2008 were reported 55260 cases from 16 departments, 39%

were confirmed cases, 69.1% were acute cases, 49.9% were male and the median age

was 11 years. The incidence rate in the area of forest was greater than in the highland

valleys since 2004. New transmission areas have been reported in the departments of

Junin, Cusco, Ayacucho and in the highlands and forest areas of Puno.

Conclusions:

Since 2004 there is more risk of infection in the forest than in the highlands. Carrion's

disease has spread to new transmission areas on the coast, highlands and forest of Peru.

OP-17

A GENE NETWORK OF LUTZOMYIA VERRUCARUM (DIPTERA:

PSYCHODIDAE) BASED ON CYTOCHROME B GENETIC VARIABILITY

ABRAHAM G. CACERES1,2

, LEE W. COHNSTAEDT3, LORENZA BEATI

4,

AND LEONARD E. MUNSTERMANN3

1Sección de Entomología, Instituto de Medicina Tropical “Daniel A. Carrión”, Facultad

de Medicina Humana, Universidad Nacional Mayor de San Marcos, Lima, Perú 2Laboratorio de Entomología, Instituto Nacional de Salud, Lima, Perú

3Yale University School of Public Health, New Haven, Connecticut, USA

4Georgia Southern University, Statesboro, Georgia, USA

Introduction. Lutzomyia verrucarum is a principal vector of Bartonella in the Peruvian

Andes. Localized epidemics of disease implies limited movement of sand flies within

geographic regions. Furthermore, the wide distribution of L. verrucarum across the

high mountain ranges of the Andes indicates the presence of isolated populations of

sand flies. The genetic relationships and migration among L. verrucarum populations

may have important disease transmission implications. The objective of this study was

(1) to determine if L. verrucarum is structured genetically across its range, (2) to

understand the relationships between populations, and (3) to identify potential barriers

to sand fly migration.

Methods. Lutzomyia verrucarum specimens were collected in Peru at single sites from

the Amazonas, Piura, Cajamarca, Lima, and Huancavelica departments and 13 districts

in Ancash. Using mitochondrial gene sequences of cytochrome b, a parsimonious gene

network was created. This analysis connected the observed haplotypes in an association

network in order to visualize the genetic relationships and genetic distance between

populations.

Results. Gene network analysis indicated the presence of three distinct populations of

L. verrucarum corresponding to the regions of Amazonas, the eastern Andes, and the

western Andes. Sand fly populations on the western slopes of the Andes were closely

related, whereas populations on the eastern slopes were more distant. The Andes form

an apparently impassible barrier to sand fly migration.

Conclusions. Several genetically distinct L. verrucarum populations were detected

within the species range with the mitochondrial cyt b sequence; the Andes mountains

strongly limit migration among them.

Financial support. United States National Institutes of Health grants U19 AI065866,

R01 AI056254.

OP-18

ENTOMOLOGICAL SURVEILLANCE AND VECTOR CONTROL IN PERU

ELENA OGUSUKU, CARMEN CRUZ, MARLENE FLORES, MIGUEL

FERNANDEZ & JAVIER HERNANDEZ

Unidad de Vigilancia y Control de Vectores. Dirección General de Salud Ambiental –

Ministerio de Salud, Lima, Peru.

Leishmaniosis and Bartonelosis are two of the most important vector borne diseases in

Peru. Both are transmitted by several species of Genus Lutzomyia. Entomological

surveillance is made by the Direcciones Regionales de Salud in all the country, where

110 profesionals are dedicated to study different vectors.

To date, more than 160 species of Lutzomyia were recorded for Peru, but only few of

them are suspected to have a role as possible vectors. The incriminated vectors for

leishmanisosis are L. verrucarum, L. peruensis, L. ayacuchensis and L. tejadai, but we

do not have incriminated species as vectors of Bartonelosis yet. We suspect that L.

verrucarum, L. maranonensis, L. peruensis have a role as vectors.

The sandflies are wide spread in the interandean valleys up to 3,500 masl, and the

Amazon basin. The major endemic areas for Bartonelosis are in the departments of

Cajamarca, Ancash, La Libertad and Cusco, while endemic areas for Leishmaniosis are

more spread in the country.

The sandfly population densities are seasonal and strongly influenced by the rainy and

dry season. There is an outbreak after the rainy season finish, the density decreases

during the dry season and increases on the beginning and the end of the rainy season in

both areas Andean valleys and Amazon basin.

In Andean valleys the sandfly behavior is strongly intradomiciliar, the residual spraying

is the main choice for vector control and very effective to prevent transmission of both

Leishmaniosis and Bartonelosis, but quite different in the jungle, were the sandflies are

extradomiciliar and the transmission usually occurs outside the villages. The control

strategy for Andean villages involve the chemical sandfly control using pyrethroid

insecticides at least twice a year in endemic areas.

OP-19

LEISHMANIASIS PROFILE IN PERU

ALEJANDRO LLANOS-CUENTAS 1,2

Universidad Peruana Cayetano Heredia (1 School of Public Health, Instituto de Medicina

Tropical Alexander von Humboldt).

Tegumentary leishmaniasis (TL, includes cutaneous, CL, and mucocutaneous, ML,

forms) is a serious health problem in Peru and in many regions of the country it is

perceived as the main health issue. On average, 10000 new cases of TL are reported per

year, due to underecords, these numbers could double. The occurrence of the disease in

the last three decades is similar to the one observed at the beginning of the XX century,

a similar trend is observed in many Latin American countries. 88% of Peruvian territory

have leishmaniasis, the highest incidence is the south-east region (departments of

Madres de Dios, San Martín, Amazonas, Húanuco and Cusco). Both children and adults

are affected in similar proportion. CL to ML ratio varies between 8.6-18.8:1 during the

last 10 years, but it was stable during the last 5 years (16.6-18.8%). Because the

surveillance system improved in the country, the recording of cases, specially the ML is

better. The number of ML cases is stable during the last 5 years and probably the rate of

ML is less than 5%.

Five Leishmania species has been described in Peru. L. (V.) braziliensis is the more

common species in the south east region (the highest endemic area), L. (V.) peruviana is

the prominent species in the Andean region (north and central western areas) and L. (V.)

guyanensis is common in the jungle (central east areas). Lower number of cases infected

by L. (V.) lainsoni and L. (M.) amazonensis was found in jungle areas. Only 5 cases of

leihmaniasis diffuse have been described in the country and L. (V.) braziliensis and L.

amazonensis were the infected species. Lbb, Lbp and Lbg were isolated from lesions of

ML patients, however, the majority of ML cases are from the jungle areas. Majority of

the studies on vectors were made in Andean regions. The main species associated with

the human transmission, using epidemiological criteria, were Lu. peruensis, Lu.

verrucarum, Lu. ayacuchensis and Lu. tejadai.

Two are the most important patterns of transmission in CL: (i) when the vector lives

outside of the dwellings, then humans are infected when they enter the natural endemic

foci, usual in Amazon regions and (ii) when the vector co inhabits with humans (intra

and/or peridomiciliary transmission), a pattern usually observed in Andean regions. In

both cases, the exposed population is generally poor and is forced to be exposed to the

infection. The first pattern is responsible for the great epidemic outbreaks and generally

associated to serious social problems such as extreme poverty, migration, wars, etc. The

young adults are mainly infected. In Peru, this has been seen in the migratory patterns of

people from the high-altitude Andean regions to jungle areas. The origin communities

are localized in interandean valleys (poor lands with limited accessibility and

expansion). The illiteracy rate in these populations in above 12% and functional

illiterates constitute 80%, thus contributing to the low work opportunities in the area.

The cheapest alternative, though with an inherent risk of infection (the great majority is

aware of this, being CL one of the most frequent) is informal work in marginally

explored jungle regions. The most common occupations are agriculture (deforestation is

a common practice, with the intention to expand arable land), informal riverside gold

mining, lumbering and harvesting natural produce (usually chestnuts). The second

pattern of transmission is responsible for the endemicity of cutaneous leishmaniasis and

occurs more frequently in the occidental interandean valleys, where, due to

geographical characteristics, climate and water availability, co inhabit in a relatively

reduced territory, humans, reservoirs and vectors. The main economic activity is

agriculture for self-sustenance. In these areas, a high percentage of localities have

difficult access to health services or complete lack thereof. Poverty percentage is high

and illiteracy rates are higher than the national average (12%), with an overall poor

presence of the State and government. Efforts to control this disease in the 60s and 70s

using only pesticides (DDT) demonstrated that the effect was transitory with no impact

in the long term. TL frequently has been associated to poverty, due to the fact that the

highly endemic countries have also high poverty rates.

OP-20

VERRUGA PERUANA

CIRO MAGUIÑA-VARGAS

Instituto de Medicina Tropical Alexander von Humboldt, Universidad Peruana

Cayetano Heredia.

The Bartonella spp. are now 25 species, most of them were isolated from animals like

cats, cows and others, being one of the main emerging diseases in the world. The first

described and ancient species is Bartonella bacillificormis, a gram negative and

polymorphic germ, it is the causative of two types of clinical syndromes: an acute initial

phase that is characterized by fever with anemia (Fiebre de la Oroya), and a chronic

phase known as “verruga peruana” with eruptions in the skin. The main vector of this

disease is Lutzomyia verrucarum, but other species like L. peruensis make this role in

Cuzco.

In Peru, the endemic areas of B. bacilliformis infections are the Inter Andean valleys at

altitudes from 500 to 3200 masl, in the last 15 years new areas of B. bacilliformis

appeared in the high jungle of Ecuador and Colombia.

The main reservoir of B. bacilliformis is apparently infected human beigns, no wild or

domestic animals were found infected with B. bacilliformis.

The acute fever phase is successfully treated with ciprofloxacine, amoxicilline, and

chloramphenicol, and the chronic phase with azitromicine and riphampicine.

El Niño phenomen caused cyclical changes in the weather of B. bacilliformis endemic

areas, these changes determined important outbreaks of more than 11000 casses in

2004, and 1300 cases in 2008.

SESSION IV: Ecology of leishmaniosis

OP-21

ECO-EPIDEMIOLOGY OF AMERICAN CUTANEOUS LEISHMANIASIS IN

ARGENTINA

SALOMÓN ODa, QUINTANA MG

b, ROSA JR

c, ACARDI SA

d

a: Ministry of Health/CONICET,

b: National University of Tucumán (UNT),

c: National

University of NorthEast (UNNE), d: National University of Misiones (UNaM),

REDILA: National Network for the Research of Leishmaniasis in Argentina

American Cutaneous Leishmaniasis (ACL) in endemic in Argentina from the northern

border to 38° SL. There are 7000 ACL cases reported since the first outbreak in 1985.

Leishmania braziliensis was isolated from cases associated with outbreaks, L.

guyanensis and L. amazonesis were also reported. Natural infections were found (PCR)

in Lutzomyia neivai, Lu. whitmani and Lu. quinquefer.

The eco-epidemiology of ACL based on phlebotomine distribution in the 9 endemic

provinces (over 150,000 sand flies) has being studied in three scales of time and space:

1) Capture site/day-week: Source forest populations can colonize the peridomestic

environment (local extinguishable populations). The distribution of peridomestic Lu.

neivai varies according to the spatial distribution of food, ecotone and wooded patches.

2) Epidemic foci/month-year: three scenarios were described including the 19 studied

outbreaks: 2a) Forest transmission/cases, associated with activities within the woods

(Lu. whitmani in NE). 2b) Forest transmission/peridomestic cases, associated with

landscape modification as deforestation-human settlement or local overflow (Lu.

neivai). 2c) Peridomestic transmission/cases, associated with adaptation of vector

species to modified environments (Lu. neivai).

3) Regional/decades: ACL epidemic wave started in the 80’s concurrent with the

appearance or prevalence of Lu. neivai. Further, transects from primary forest to

villages in the hyper-endemic area had increasing relative and total abundance of Lu.

neivai, and increasing ACL incidence. In the central dry region (Chaco) the scattered or

family clustered ACL cases are still associated with Lu. migonei without Lu. neivai.

Phlebotomine are better indicators in time and space of ACL transmission than human

ACL data from secondary sources. In Argentina the relative and absolute abundance of

Lu. neivai and Lu. migonei could be used for surveillance and monitoring the ACL

outbreak risk. Recommendations of control were developed for each scale and scenario.

OP-22

ECO-EPIDEMIOLOGY OF VISCERAL LEISHMANIASIS IN ARGENTINA

SALOMÓN ODa, ACARDI SA

b, ROSA JR

c, QUINTANA MG

d

a: Ministry of Health/CONICET,

b: National University of Misiones (UNaM),

c: National University of NorthEast (UNNE),

d: National University of Tucumán

(UNT), REDILA: National Network for the Research of Leishmaniasis in Argentina

From 1925 to 1989 14 cases of leishmaniasis with visceral involvement were reported

from Argentina. They were scattered through the area of cutaneous leishmaniasis,

without records of Lutzomyia longipalpis or other Leishmania chagasi known vectors.

Lu. longipalpis were reported in 1953 (Candelaria) and in 2000 (Corpus) in the

northeastern province of Misiones, without visceral leishmaniasis (VL).

Since the reports of Lu. longipalpis-VL in Campo Grande (Brazil) and Asunción

(Paraguay), phlebotomine surveillance was intensified in the northern border of

Argentina. Lu. longipalpis was found in Clorinda (Formosa province) on the border

with Asunción in 12/2004. The city was screened again for phlebotomine in 11/2007,

with traps in 140 peridomestic sites. Lu. longipalpis was found in 7 sites (n=51), in the

two neighbors already positive in 2004. Canine VL, but not human VL, is spreading all

over Clorinda despite the Lu. longipalpis clustered distribution. Infected dogs came also

through the border.

The first human autochthonous case, with concurrent canine VL and Lu. longipalpis,

was recorded during 5/2006 in Posadas (Misiones province). Lu. longipalpis (n=5064)

was found in 42% of the 314 sites sampled (‘worst scenario’ in 400 x 400 m, 2-3/2007),

but only 5.2% of these sites had more than 31 Lu.longipalpis/trap. No spatial clustering

of positive traps was found. L. infantum chagasi was typified from human and dogs. Up

to 5/2008 VL was reported in 19 humans (3 dead), and estimated in 3500 dogs.

La Banda (Santiago del Estero province) reported 4 human VL cases and 8 dogs with

VL (screening around the cases) from 10/2007 to 5/2008. All the cases were in an area

of 1.5 km2. The focus was sampled in 11/2007 and 4/2008 (134 Lu. migonei, 14 Lu.

cortelezzii, 2 Lu. neivai) with 15-21 traps up to 250 m from each case. The human case,

the dogs and the highest abundance of Lu. migonei were concurrent (10 m). L. chagasi

was typified from VL human cases. The role of Lu. migonei as a permissive vector of

VL is under research.

OP-23

SYMPATRY OF INFECTED LUTZOMYIA LONGIPALPIS AND LUTZOMYIA

EVANSI IN A FOCUS OF AVL IN THE SEMI-ARID REGION OF WESTERN

VENEZUELA

NÉSTOR AÑEZ1*

, ERSY VARGAS2, AGUSTINA ROJAS

1, VICENTE

MEDINA2, GLADYS CRISANTE

1, JOSÉ Y. YÉPEZ

2

1: Universidad de Los Andes, Facultad de Ciencias, Departamento de Biología, 5101,

Mérida, Venezuela

2: Universidad Nacional Experimental “Francisco de Miranda”, Coro, Venezuela

A total of 115 specimens of Lutzomyia longipalpis (90) and Lu. evansi (25) were

captured in an active focus of American Visceral Leishmaniasis (AVL) in a village

located at the semi-arid region of western Venezuela. Both sand fly species were caught

using CDC traps placed in a goat yard, 10 m apart from a house where an acute case of

AVL was detected and diagnosed by clinical, sero-parasitological and molecular (PCR)

methods. The sand flies were dissected and identified accordingly. The digestive tracts

were processed for PCR assay to detect Leishmania infantum specific DNA. The results

revealed infection with L.infantum in 4 out of the 90 (4.4%) processed guts of Lu.

longipalpis, and in 12% (3/25) of Lu. evansi. The high infection range detected in both

species of flies appears to correspond with the high level of seropositivity to

Leishmania-infection from people (22.5%) and dogs (87.5%) sampled at the same

village. The present results suggest that Lu. longipalpis and Lu. evansi are sympatrically

distributed in the semi-arid, and both species are efficient vectors of L.infantum in this

part of western Venezuela.

Acknowledgements: Financial support given by CDCHT-ULA and FONACIT –G-

2005000370 (NA) .

OP-24

STUDY ON THE VECTORIAL COMPETENCE OF LUTZOMYIA ALMERIOI

GALATI & NUNES, 1999 FOR TREE SPECIES OF LEISHMANIA: L.

(LEISHMANIA) INFANTUM CHAGASI (CUNHA & CHAGAS, 1937), L.

(VIANNIA) BRAZILIENSIS (VIANNA, 1911) AND L. (L.) AMAZONENSIS

(LAINSON & SHAW, 1972)

M. F. C. SANTOS1, V. L. B. NUNES

2, E. A. B. GALATI

3, A. R. O. ANDRADE

1, A.

P. VIEIRA2, M. E. G. ROCCA

4, R. B. AQUINO

4, C. C. P. ARRUDA

1.

1Centro de Ciências Biológicas e da Saúde, UFMS, Campo Grande, Brazil.

2Centro de Ciências Biológicas, Agrárias e da Saúde, UNIDERP, Campo Grande,

Brazil. 3Faculdade de Saúde Pública, USP, São Paulo, Brazil,

4Núcleo de Controle de Zoonoses, Prefeitura do Município de Bonito, Bonito, Brazil.

It was investigated the vectorial competence of Lutzomyia almerioi, for three species of

Leishmania: L. (Leishmania) infantum chagasi, L. (Viannia) braziliensis and L. (L.)

amazonensis, by means of research of experimental infection, exposure of wild caught

females to hamsters infected with each one of the cited leishmanias or infected dog and

attempt of transmission tests by bite in non infected hamsters. The specimens of Lu.

almerioi had been captured in the entrance and the interior of cave (20o

52’10” S; 56o

35’06” W) in the Pitangueiras Farm, Planalto da Bodoquena. Three experiences had

been made, in March and December of 2006 and March of 2007. It was possible to

confirm, in optical microscope and PCR, that Lu. almerioi can be infected, in laboratory

conditions, by L. (L.) infantum chagasi, L. (V.) braziliensis and L. (L.) amazonensis.

PCR also confirmed, that this species is capable to transmit, by bite, L. (V.) braziliensis

and L. (L.) amazonensis to clean hamsters. It is concluded, that Lu. almerioi is vector

for L. (V.) braziliensis and L. (L.) amazonensis in the region of Planalto da Bodoquena,

specially in cave areas, where this phlebotomine is particularly abundant. The

transmission of L. (L.) infantum chagasi to hamster was not detected, thus, for the used

criteria, the species still cannot be considered vector of L. (L.) infantum chagasi in that

region. These results bring useful information for the improvement of the monitoring

epidemiologic questions of leishmaniasis in the region.

OP-25

SAND FLIES FAUNA AND BIOGEOGRAPHICAL ASPECTS OF VISCERAL

LEISHMANIASIS, IN NORTHEAST REGIONAL BELO HORIZONTE, MINAS

GERAIS – BRAZIL

LARA SARAIVA1, DEBORAH APARECIDA CARVALHO

2, CRISTIANI

CASTILHO SANGUINETTE2, CAMILA RAGONEZI GOMES

1, CARINA

MARGONARI SOUZA2, CHRISTIAN FREITAS REZENDE

1, JOSÉ

DILERMANDO ANDRADE FILHO2, ALDA LIMA FALCÃO

2, MARIA NORMA

MELO1.

1-UFMG,

2-CPQRR

Introduction: Belo Horizonte is the Brazilian city with high population which suffers

more with the occurrence of VL. From 1994 to 2007, 891 human cases of the disease

were notificated, 22 deaths in the last three years and high rates of canine prevalence.

Northeast Regional presents the highest historical average of human cases of VL.

Objectives: to study the seasonal variation of sand flies fauna and to describe the

environmental situations through geoprocessing analysis and the peridomestic

characterization. Methodology: fortnightly entomological captures were carried out

during one year using HP light traps, in 15 places following the location of human cases

of VL in 2005. All collected sand flies were processed and identified. The

environmental characterization was carried out through a filled form and photographs.

The geoprocessing analyzed the influence of vegetation, hydrograph, altitude and

poverty pockets in the occurrence of human cases of VL, canine cases of leishmaniasis

and sand flies vectors. Results: A total of 633 sand flies belonging to the genera

Lutzomyia were captured, L. whitmani accounted for 75% and L. longipalpis 11%,

together responding to 85% of the total. The majority of the peridomestic places have

shown inadequate hygienic care conditions, allowing the development of sand flies. The

geoprocessing analysis reveal no relevant correlation between biogeographical aspects

and human and canine cases of leishmaniasis and the presence of sand flies. Both

human and canine cases occurred in the areas with low average of sand flies occurrence.

Conclusion: This study confirms the urbanization of the L. longipalpis and suggests the

same pattern to L. whitmani. The biogeographical analysis suggested that the

peridomestic places are important to keep the sand flies populations in urban areas, and

pointed out a new profile of occurrence not related to classic factors.

Acknowledgements: Community of Northeast Regional .

Financial Support: LeishEpinetSA-(EEC), FIOCRUZ, FAPEMIG.

OP-26

EMERGING CUTANEOUS LEISHMANIASIS IN ISRAEL

ALON WARBURG1, ROI FAIMAN

1, IBRAHIM ABBASI

1, ABEDELMAJEED

NASEREDDIN1, LIONEL SCHNUR

1, RUBEN CUNIO

1, MILENA

SVOBODOVA2, JAN VOTYPKA

2, PETR VOLF

2, CHARLES L. JAFFE

1

1

The Hebrew University of Jerusalem, Jerusalem, Israel; 2Charles University, Prague,

Czech Republic

In Israel cutaneous leishmaniasis (CL) is emerging in several new foci. Between 2001-

2003, 75 CL cases from foci near Tiberias were shown to be caused by two different

Leishmania tropica strains. Parasites from the northern focus were antigenically similar

to L. major and Phlebotomus (Adlerius) arabicus was incriminated as their vector with

5% infected females. In the southern focus P. (Paraphlebotomus) sergenti (10-20%

infected) was the only vector. In experimental infection studies, parasites from the

northern focus developed only in P. arabicus while parasites from the southern focus

infected both Phlebotomus species. Like other permissive vectors, the luminal surface

of P. arabicus midgut epithelium was heavily o-glycosylated while that of P. sergenti

was not. Rock hyraxes (Procavia capensis) were the most prominent mammals in both

foci and L. tropica DNA was detected in 14 out of 121 hyraxes. Between 2006-2008,

72 CL cases from rural areas near Beit She’an, 30 km south of Tiberias, were diagnosed

as caused by L. major. Nine of 40 voles (Microtus guentheri) collected near patients’

homes were identified by PCR as L. major, and P. papatasi, the known vector of L.

major, was the most abundant sand fly species in the area. Emergence of new CL foci

in Israel is probably caused by the encroachment of hyraxes upon human habitation (L.

tropica) and the proliferation of voles in agricultural fields near villages (L. major). The

adaptation of parasites to new, highly-susceptible vectors (L. tropica/P. arabicus) or

host species (L. major/M.guentheri), also plays a role.

OP-27

MAPPİNG OF RİSK AREAS FOR VİSCERAL AND CUTANEOUS

LEİSHMANİASİS RELATED WİTH DİSTRİBUTİON OF VECTOR SPECİES

İN WESTERN PART OF TURKEY USİNG GEOGRAPHİC INFORMATİON

SYSTEMS

YUSUF OZBEL1, I. CÜNEYT BALCIOGLU

2, KİRAMİ ÖLGEN

3, FATİH

SIMSEK4, SERAY OZENSOY TOZ

1, HATİCE ERTABAKLAR

5, SAMİYE

DEMIR6, M. ZİYA ALKAN

1

1Ege University Medical School Department of Parasitology, Bornova, Izmir, Turkey

2Celal Bayar University Medical School Department of Parasitology, Manisa, Turkey

3Ege University Faculty of Letters Department of Geography, Bornova, Izmir, Turkey

4Adnan Menderes University Science Faculty Department of Biology, Aydin, Turkey

5Adnan Menderes University Medical School Department of Parasitology, Aydin,

Turkey 6Mustafa Kemal University Science Faculty Department of Biology, Hatay, Turkey

Leishmaniasis are present in two clinical forms, visceral and cutaneous, in Turkey.

While the number of recorded visceral leishmaniasis (VL) cases is around 50-60 and the

disease is seen in every region of Turkey, the number of recorded cutaneous

leishmaniasis (CL) cases is more than 2000 and the disease is getting spreading

throughout the country including western part. The aim of the present study was to carry

out entomological survey and to produce risk maps related with probable vector species

for western part of Turkey.

For determining the vector species, an entomological survey was carried out in

Kusadasi town and Aydin province where human and canine visceral leishmaniasis and

CL endemic region. The study area was 48x88 km2. The area was divided 66 squares as

16 km2 each and at least one location was chosen in each square during the field work.

The detailed ecological information was also collected for each location.

The results of entomological studies were shown that the probable vector species are

Phlebotomus tobbi and P. neglectus for VL, P. similis for CL in this western

leishmaniasis focus. The maps were produced to show the distribution of vector species

in the study area using geographic information system (GIS) and then the risk maps

were developed based on known distribution of these three species for the western part

of Turkey. Methodology of the development of the risk maps was based on weighted

score analysis. Altitude, aspect, Normalized Difference Vegetation Index (NDVI) and

Land Surface Temperature (LST) values were used as parameters of the analysis.

Altitude and aspect were derived from SRTM data set, NDVI and LST values were

calculated from Landsat TM data of the study area.

The risk maps were shown the potential distribution areas of probable vector species

according to the parameters used and the use of GIS allowed the identification of classes

of leishmaniasis risk that may be useful information to guide control program

interventions.

The study is supported by EU Integrated Project - 010284 (EDEN).

SESSION V: Detection of Leishmania in sandflies. Other microorganisms in

sandflies

OP-28

DETECTION AND IDENTIFICATION OF LEISHMANIA PARASITES IN

SAND FLIES COLLECTED IN IRAQ AND AFGHANISTAN.

RUSSELL E. COLEMAN,1 LISA P. HOCHBERG,

1 JOHN L. PUTNAM,

2

KATHERINE I. SWANSON,1 JOHN S. LEE,

3 JAMES C. MCAVIN,

2 JOHN K.

MOULTON,4 MONICA L. O’GUINN,

3 & MICHAEL K. FAULDE

5

Affiliation: 1 Department of Entomology, Walter Reed Army Institute of Research,

Silver Spring, MD; 2

Epidemiological Surveillance Division, Air Force Institute of

Operational Health, San Antonio, TX; 3

Virology Division, U.S. Army Medical

Research Institute of Infectious Diseases, Fort Detrick, MD; 4 Department of

Entomology and Plant Pathology, University of Tennessee, Knosvill, TN; 5

Department

of Medical Zoology, Central Institute of the Bundeswehr Medical Service, D-56065

Koblenz, Germany

Abstract In this talk we provide an overview of efforts to detect Leishmania parasites

in sand flies collected in Iraq and Afghanistan from 2003-2006. A total of 169,594 sand

flies were collected, with 7,174 pools containing 60,790 sand flies tested for the

presence of Leishmania parasites using a Leishmania-generic real-time PCR assay. A

total of 620 pools were considered true positives, with an infection rate of 1.02%. We

subsequently sequenced a 360 base-pair region of the Glucose-6-phosphate-isomerase

gene from 739 pools of sand flies. These pools included 570 PCR-positive samples,

158 PCR-indeterminate samples, and 11 PCR negative samples. Sequencing indicated

that 34 (4.6%) of these 739 samples contained medically-important species of

Leishmania, to include 19 samples containing L. donovani complex parasites, 9

containing L. major, 4 containing L. tropica, and 2 containing a parasite that appeared

similar to both L. major and L. tropica. A total of 12 of the 19 L. donovani complex-

parasites were subsequently determined to be L. infantum. Three hundred and forty

(46.5%) samples contained L. tarentolae DNA, while no Leishmania DNA was detected

in 365 samples (49.8%). The implications of these findings are discussed.

OP-29

CUTANEOUS LEISHMANIASIS CAUSED BY LEISHMANIA INFANTUM AND

TRANSMITTED BY PHLEBOTOMUS TOBBI

JAN VOTÝPKA, MILENA SVOBODOVÁ, BULENT ALTENa, PETR VOLF

Department of Parasitology, Faculty of Science, Charles University, Prague, Czech

Republic, e-mail: vapid@natur.cuni.cz aDepartment of Biology, Hacettepe University, Ankara, Turkey

In Turkey, both Leishmania tropica and L. infantum are endemic in several regions.

Cutaneous leishmaniasis (CL) had been assigned almost exclusively to L. tropica. Our

study in Cukurova region (South Anatolia, Turkey) revealed new CL focus caused by

Leishmania infantum. Since 1985, thousands of new CL cases have emerged there and

according to the local health centers, hundreds of human cases continue to occur every

year. Small, non-ulcerating lesions prevailed and patients were negative in rK39 tests

for antibody detection for human VL by DiaMed-IT LEISH immuno-chromatographic

dipstick test. The most abundant sand fly species, Phlebotomus tobbi, was found

positive for Leishmania promastigotes with a prevalence of 1.4% (13 out of 898

dissected females). The isolated strains from sand flies were identical with those

obtained from patients with CL and were typed by PCR-RFLP and MLST (for icd, me,

mpi, gpi and fh genes) techniques as L. infantum. Phylogenetic analysis revealed

similarity to MON-188 and a clear difference from the MON-1 clade and other

viscerotropic Turkish strains of L. infantum. Blood-meal identification based on

sequencing of the amplified part of the cyt b gene was found to be a very sensitive

method with an efficiency level of almost 85% and showed that P. tobbi feeds

preferentially on cattle (approx. 70%) and humans (approx. 10%). On the other hand, no

dog blood was found at all. Furthermore only human blood was detected in four

Leishmania-infected females of P. tobbi. These findings, together with the high number

of CL patients and relative scarcity of dogs in the focus, suggest that the transmission

cycle is anthroponotic.

OP-30

TWO NEW SPECIES OF GREGARINES (APICOMPLEXA) FROM

PHLEBOTOMUS SERGENTI AND P. TOBBI (DIPTERA, PSYCHODIDAE)

LUCIE LANTOVA, MILENA SVOBODOVA, JAN VOTYPKA and PETR VOLF

Department of Parasitology, Faculty of Science, Charles University, Vinicna 7, Prague

2, 128 44 Czech Republic, e-mail: lantova@centrum.cz

Gregarines (Apicomplexa) are parasites of invertebrates found mostly in the gut or in the

body cavity. Those occurring in mosquitoes and sand flies, genus Ascogregarina, are

thought to belong to order Eugregarinorida as they do not possess merogony in their

life cycle. We studied new gregarine species from P. sergenti and P. tobbi and

compared their features to already described Ascogregarina chagasi from Lutzomyia

longipalpis. Significant differences were observed in the length of gamonts,

gametocysts and oocysts of gregarines as well in their development. The life cycle of

gregarine from P. tobbi was similar to that of A. chagasi as its sexual stages occurred in

both sand fly sexes. In contrast, gregarine from P. sergenti developed sexual stages

exclusively in blood fed females. Experimental infections of various sand fly species

revealed that gregarines are highly host-specific. Gregarine from P. sergenti produced

oocysts only in its natural host, not in the other sand fly species tested (P. papatasi and

P. arabicus). Gregarine from P. tobbi developed oocysts only in three out of 260 adults

of P. perniciosus and did not complete its life cycle in P. sergenti. The life cycle of

gregarine in larvae and adults of P. sergenti was documented using histology (PAS

reaction followed by Ehrlich´s hematoxylin) and electron microscopy. The gregarine

had a major pathogenic effect on the insect host; infection of P. sergenti significantly

decreased the number of adults emerging from pupae and increased mortality of males

and females. In contrast, there was no effect on fecundity. Phylogenetic study of SSU

rDNA revealed that gregarines from sand flies differ from those parasitizing

mosquitoes, therefore we propose the erection of the new genus Phlebogregarina. In

addition, both sand fly and mosquito gregarines are closer to neogregarines, a group that

possesses merogony in their life cycle.

OP-31

EPIDEMIOLOGY OF LEISHMANIASIS IN ISRAEL: IMPLEMENTATION OF

MOLECULAR TOOLS FOR LEISHMANIA DETECTION AND HOST BLOOD

MEALS IDENTIFICATION.

VALINSKY L., KHALFA Z., AND ORSHAN L.

Central Laboratories, Ministry of Health, Jerusalem, Israel.

Cutaneous leishmaniasis has recently expanded its geographical distribution in Israel.

As part of the governmental efforts to understand the epidemiology of the disease and

employ effective intervention, we developed and applied new molecular methods to

detect Leishmania in pooled sand flies and to identify the blood source in engorged

females.

Screening large numbers of sand flies is crucial to reveal high risk locations and seasons

for disease transmission. Thus we developed a high throughput sensitive molecular

method for the parasite DNA detection in pools of 20 females in locations where one

Phlebotomus species predominates. Since 2005, approx. 38,000 female sand flies were

tested for Leishmania DNA presence by ITS1 PCR and RFLP. In the town of Ma'ale

Adummim, 7.8% (18/217) of the pools tested were found positive in 2005, one year

after the noteworthy L. tropica outbreak. In 2006 infection rates decreased to 2.4%

(19/791) and further decreased to 0.6% in 2007 (3/479). In the nearby village of Kfar

Adummim, Leishmania infections in sand flies were around 8% in 2005 and 2006, and

decreased to 3% in 2007. In 2007 when L. tropica infection rates in sand flies from the

Judean Desert were low, high infection rates (29%) were obtained in Tiberias in the

northern Jordan Valley. In 2007 we also sampled sand fly specimens from one

emerging L. tropica focus in the Samaria Mountains and two endemic L. major foci in

southern Israel. High prevalence was detected in samples from a military region in the

western Negev Desert.

We developed a PCR-based method to define vector-host preferences regardless of prior

knowledge of host occurrence. Primers were designed to amplify mitochondrial 12S and

16S ribosomal genes of vertebrates, and not to amplify arthropod DNA. Blood meal

identification was conducted by analyzing the DNA sequence of species-specific, non-

conserved regions of the amplified fragment.

We identified the host animals of 311 engorged female Phlebotomus sand flies collected

in the Judean Desert and Tiberias during 2005-2006. Co-analyzing the host preferences

and Leishmania infection results enabled us to identify the reservoir animal, the rock

hyrax, with a high degree of confidence. This method of blood meal identification is

universal and can be applied to any blood-sucking arthropod.

The spatial and temporal distribution of the parasite prevalence and host preferences in

sand flies are discussed.

OP-32

NATURAL INFECTION OF SAND FLIES BY LEISHMANIA SPP. IN

NORTHEAST REGIONAL , BELO HORIZONTE, MINAS GERAIS – BRAZIL.

LARA SARAIVA1, DEBORAH APARECIDA DE CARVALHO

2, CÉLIA MARIA

FERREIRA GONTIJO2, JOSÉ DILERMANDO ANDRADE FILHO

2, ALDA

LIMA FALCÃO2, MARIA NORMA MELO

1.

1-UFMG,

2-CPQRR

Introduction: Over the last 20 years, an increase of the number of human cases of

visceral leishmaniasis (VL) occurred in Brazil. VL are considered emergent in some

areas and reemergent in others. The control measures of visceral leishmaniasis in Brazil

are inefficient, even in municipalities with a huge program of euthanasia of serological

positive dogs and insecticide spraying in areas with high incidence rates of human and

canine cases, like occurs in Belo Horizonte city. The knowledge of the natural infection

rates of sand flies and the species of Leishmania infecting them can improve the control

measures. Methodology: We carried out fortnightly entomological captures during one

year using HP light traps in 15 places following the location of human cases of VL in

2005. Captures using Shannon light trap were performed out in places where infected

sand flies were found. The females were identified and submitted to DNA extraction

and PCR and PCR-RFLP reactions to verify the natural infection rate by Leishmania

sp., and the identification of the parasite species. Results: A total of 633 sand flies

belonging to the genera Lutzomyia were captured. The natural infection rates were: Lu.

longipalpis 19%, Lu. whitmani 3,8% and Lu. termitophila 33,3% all of them with Le.

chagasi. The females of the cortellezzi complex present 3,2% of natural infection rate

with Le. braziliensis and 3,2% Leishmania sp. Lu. intermedia presents 14,3% with

Leishmania sp. Conclusion: This study shows the important circulation of Le. chagasi

in that Regional, even species that have never been infected with Leishmania presented

infection with Le. chagasi.

Acknowledgements: Community of Northeast Regional.

Financial Support: LeishEpinetSA (EEC), FIOCRUZ, FAPEMIG.

SESSION VI: Leishmania-sandfly interaction

OP-33

SAND FLY PERITROPHIC MATRIX AND ITS ROLE IN LEISHMANIA

DEVELOPMENT

JOVANA SADLOVA and PETR VOLF

Department of Parasitology, Charles University, Vinicna 7, 128 44 Prague 2, Czech

Republic e-mail: JovanaS@seznam.cz

The peritrophic matrix (PM), an acellular chitin-containing envelope separating the gut

lumen from the midgut epithelium, was investigated in Phlebotomus duboscqi females.

Light microscopy of native preparations, histology and electron microscopy revealed

that P. duboscqi belongs to sand fly species with fast PM development. The formation

of PM had several distinct stages. Secretion of electron lucent fibrils, presumably chitin,

started immediately after the bloodmeal ingestion. About six hours later, secretion of

amorphous electron-dense components, presumably proteins and glycoproteins,

prevailed. The PM matured in less than 12 hours, it is about 2µm thick and consists of

thin laminar outer layer and thick amorphous inner layer. The thickness of PM

increased to about 5µm during next two days.

No differences were found in timing of assembly and degeneration of the PM in

Phlebotomus duboscqi females infected by Leishmania major. In both groups, infected

and uninfected ones, the disintegration of the PM started at its posterior end. Native

observation of GFP promastigotes showed that although parasites were present in high

densities in the anterior part of blood meal bolus they escaped from the PM on the

posterior end only. Additionally, electron microscopy did not reveal the ability of

promastigotes to penetrate through the PM although it was observed that yeasts

contaminating P. duboscqi midgut were able to lyse and disrupt the PM. These results

suggest that L. major chitinase does not have an important role in parasite escape from

the peritrophic sac. Promastigotes stay in intraperitrophic space until PM is broken by

sand fly-derived chitinases and only then migrate anteriorly. A novel role is ascribed to

anterior plug, the part of the PM secreted by thoracic midgut (TGM); it functions as a

temporary barrier which stops a forward migration of nectomonads to the TMG until its

disintegration.

Disintegration of the PM tightly coincided with morphological transformation of

parasites from stumpy procyclic forms to long nectomonads. We suppose that this

process is triggered by contact with saliva components ingested into the midgut.

OP-34

DEMONSTRATION OF A LEISHMANIA SEXUAL CYCLE IN THE SAND FLY

VECTOR

PHILLIP LAWYER1, NATALIA AKOPYANTS

2, NICOLA KIMBLIN

1,

RACHEL PATRICK1, NAGILA SECUNDINO

1, STEPHEN BEVERLEY

2, AND

DAVID SACKS1

1Laboratory of Parasitic Diseases, NIAID, NIH, Bethesda, MD 20892

2Dept. of Molecular Microbiology, Washington University School of Medicine, St.

Louis, MO 63110.

The clinical outcome of Leishmania infection, ranging from localized cutaneous lesions

that heal spontaneously, to visceral dissemination that is fatal in the absence of

treatment, is determined in large part by the species of Leishmania transmitted by their

respective sand fly vectors. The considerable inter- and intra-species diversity of

Leishmania is thought to have arisen by gradual accumulation of divergent mutations,

and sexual recombination is not thought to have significantly impacted on the

population structure of these organisms. Based on the strong linkage disequilibrium

displayed by Leishmania species, these parasites are thought to be essentially clonal.

This notion must be reconciled, however, with the accumulating examples of naturally

occurring hybrid strains, which appear to share phenotypic and genotypic markers from

two perceived species, providing circumstantial evidence for sexual recombination.

Clearly, the clonality vs sexuality debate would be far better informed if Leishmania

were actually shown to be capable of genetic exchange, evidence for which has been

lacking until now. By co-infecting sand flies with two parental lines of Leishmania

bearing distinct drug- resistant markers, we have explored the possibility of genetic

exchange occurring during the growth and development of extracellular stage L. major

promastigotes in the midgut of a natural sand fly vector species, Phlebotomus duboscqi.

We provide the first conclusive evidence that the invertebrate stages of Leishmania are

fully capable of a sexual cycle, generating both diploid and triploid progeny bearing

hybrid genotypes that strongly suggest meiotic division and uniparental inheritance of

maxi-circle kDNA..

OP-35

DEVELOPMENT OF LEISHMANIA INFANTUM PROMASTIGOTES

TREATED WITH AMPHOTERICIN B IN PHLEBOTOMUS PERNICIOSUS

AND LUTZOMYIA LONGIPALPIS

CARLA MAIA, LENEA CAMPINO, JOVANA SADLOVA, LUCIE JECNA and

PETR VOLF

Unidade de Leishmanioses, CMDT, Instituto Higiene e Medicina Tropical,

Universidade Nova de Lisboa, Lisboa, Portugal

Department of Parasitology, Fac. Sci., Charles University in Prague, Czech Republic

Introduction: Treatment options for visceral leishmaniasis (VL) vary between

countries but are generally limited to pentavalent antimony and amphotericin B (AMB)

as first-line drugs, pentamidine, aminosidine and the new oral agent miltefosine.

Although AMB has been widely used in VL treatment in Mediterranean basin, in

particular in Leishmania/HIV co-infections and paediatric cases, there are few studies

on AMB resistance so far.

Objective: The main aim was to study development of AMB-treated line of L. infantum

in Phlebotomus perniciosus and Lutzomyia longipalpis in order to predict the potential

of spreading the amphotericin-resistant lines in nature.

Methods: L. infantum MON-1 (MCAN/PT/05/IMT-373) wild type and AMB treated

parasite line R160 were used in this study. Line R160 was obtained by subculturing

promastigotes in media containing increasing concentrations of AMB. Colonized P.

perniciosus and L. longipalpis were membrane-fed on rabbit blood containing

promastigotes and engorged females were dissected 2 and 10 days later. The location

and number of promastigotes in the midgut was estimated under light microscope.

Results: Wild type parasites developed well in both sand flies genera producing high

infection rates and high percentage of heavy infections with frequent colonization of the

stomodeal valve on day 10 postinfection. Promastigotes treated with AMB produced

lower infection rates with majority of light infections. During late stage infections (day

10 p.i.) they did not colonize the stomodeal valve. Similar development of AMB-treated

line was observed in P. perniciosus and L. longipalpis.

Conclusions: Despite the low infection rate, promastigotes of AMB-treated line were

able to develop late-stage infections in the midgut of both sand flies species tested.

More studies are needed to determine the vectorial capacity of sand flies to transmit

AMB-resistant parasites.

Financial support: This work was supported by EU/FEDER, POCI/CVT/56357/2004

from Fundação Ciência e Tecnologia (FCT), Portugal and by the Ministry of Education

of the Czech Republic (MSM0021620828). C. Maia (SFRH /BD/12523/2003) has a

fellowship from FCT.

OP-36

NON-SPECIFIC ANTIVIRAL RESPONSE OF LUTZOMYIA LONGIPALPIS

CELL LINE TRIGGED BY DOUBLE-STRAND RNA.

PITALUGA AN1,2

, MASON PW2, TRAUB-CSEKO YM

1.

1 - Laboratório de Biologia Molecular de Tripanosomatídeos e Flebotomíneos, Instituto

Oswaldo Cruz-Fiocruz, Rio de Janeiro, Brazil.

2 - Department of Pathology, University of Texas Medical Branch, Galveston, USA

Introduction: Lutzomyia longipalpis is the principal vector of visceral leishmaniasis in

the Americas but can also transmit bartonellosis and some virus. The role of sandfly

immune system in defense is poorly understood and the mechanism underlying viral

resistance is unknown. Recently, working in a RNAi silencing system for L. longipalpis

we identified an antiviral response to a non-specific double-strand RNA transfection.

Objective: Characterization of a non-specific antiviral response in L. longipalpis.

Methods: Cultured embryonic cells were transfected with various double-stranded

RNAs (dsRNA) using West Nile virus (WNV) virus-like particles (VLPs) expressing

luciferase as the target RNA to demonstrate effective gene knock-down.

Results: When luciferase dsRNA was introduced into these cells, they produced the

expected reduction in VLP-encoded luciferase, suggesting specific silencing of the

luciferase gene. Surprisingly, we found that unrelated dsRNAs, which included those

specific for several L. longipalpis gene sequences and Escherichia coli beta-

galactosidase, diminished replication of the VLP-encoded genome.

Conclusions: These results are the first indication for a nucleic acid-induced, non-

specific antiviral response in this important insect vector.

Financial Support: UTMB, CAPES and FIOCRUZ

OP-37

SAND FLY – LEISHMANIA INTERACTION: CONCEPT OF SPECIFIC

VERSUS PERMISSIVE VECTORS

PETR VOLF, LUCIE JECNA, AND ANNA SVAROVSKA

Department of Parasitology, Charles University, Vinicna 7, 128 44 Prague 2, Czech

Republic, e-mail: volf@cesnet.cz

Two sand fly species, Phlebotomus papatasi and P. sergenti, display remarkable

specificity for Leishmania parasites that they transmit in the nature (L. major and L.

tropica, respectively). Most other sand fly species examined so far are susceptible to the

development of a broad range of Leishmania species; we call them permissive vectors.

They include species transmitting parasites of the L. donovani complex. Previous

studies showed that Leishmania -sand fly interactions in specific vectors are mediated

by parasite surface glycoconjugate lipophosphoglycan (LPG) and sand fly lectins. Our

work on L. infantum / L. major hybrids has confirmed these findings: hybrids

possessing L. major LPG were able to develop in P. papatasi. On the other hand, we

showed that interactions involving permissive sand fly species utilize another

molecules. We found that LPG, essential for the attachment of Leishmania major in the

specific vector P. papatasi, was not required for the parasite adherence or survival in the

permissive vectors P. perniciosus, P. arabicus, and Lutzomyia longipalpis. Instead,

binding in several permissive vectors correlated with the occurrence of a lectin-like

activity on the parasite surface and with the presence of sand fly midgut glycoproteins

bearing N-Acetyl-galactosamine. This new binding modality has important implications

for co-evolution of the parasite with the vector. It enables the successful transmission of

Leishmania in new vectors. Two good examples are transmission of atypical L. tropica

by P. arabicus, and adaptation of Mediterranean L. infantum in the New World sand fly

Lutzomyia longipalpis. The second event led to the establishment of visceral

leishmaniasis in Latin America.

OP-38

SEX PHEROMONE GLANDS OF LUTZOMYIA LONGIPALPIS (DIPTERA:

PSYCHODIDAE) MALES: MORPHOLOGICAL ALTERATIONS DURING

DEVELOPMENT

CAROLINA N. SPIEGEL1, SANDRA M. P. OLIVEIRA

2, ALEXANDRE A.

PEIXOTO2 & MAURILIO J. SOARES

3

1Universidade Federal Fluminense;

2Instituto Oswaldo Cruz;

3Instituto Carlos Chagas

The sand fly Lutzomyia longipalpis (Diptera: Psychodidae) is the main vector of

American Visceral Leishmaniasis. Adult L. longipalpis males produce terpene-rich sex

pheromones with dual function of male aggregation and sex pheromone. These

pheromones are released to the environment from papular cuticular structures located in

pale spots on the third and/or fourth abdominal tergites, depending on the insect

population. The secretion is produced by pheromone gland cells grouped beneath the

cuticle, connected to the exterior via a small cuticular duct. The large columnar

secretory cells present two distinct parts: a basal region with vacuoles and an apical

region with an end-apparatus. Ultrastructural study with L. cruzi revealed that the

vacuoles are in fact lipid droplets in close association with peroxisomes and

endoplasmic reticulum profiles, indicating the possible role of lipids in the pheromone

synthesis. To better understand the morphogenesis of these gland cells we analyzed

their fine structure by transmission electron microscopy (TEM). Analysis of 4th

abdominal tergites of 0-6 hour-old adult L. longipalpis males by TEM revealed that at

this time the gland cells are smaller than the hypertrophied epithelial cells characteristic

of older insects. Small microvilli are observed at the end apparatus, while lipid droplets

are absent or else present in small size and number. There are almost no peroxisomes,

but large numbers of mitochondria can be seen. At 9-12h after emergence a larger

amount lipid droplets can be observed, as well as several peroxisomes. At 12-14h after

emergence the lipid droplets are primarily distributed near the microvilli at the apical

portion of the gland cells, but are smaller than those in mature older males (4-day-old),

which are large and distributed throughout the gland cell cytoplasm. Thus, lipid droplets

start to appear in the gland cells cytoplasm about nine hours after adult emergence, their

number and size increasing with age. Such lipid droplets may be involved in the

pheromones biosynthesis.

Acknowledgements: CNPq, FAPERJ and FIOCRUZ

SESSION VII: Sandfly saliva

OP-39

THE INFLUENCE OF SAND FLY SALIVA ON VISCERALIZATION OF

LEISHMANIA CHAGASI

D. A. ELNAIEM1, C. MENESES

2, C. LEUTENEGGER

2, L. SOONG

3 & G. C.

LANZARO2

1. Laboratory of Vector Molecular Biology LMVR/NIAID/NIH, Washington, DC, 2.

Department of Pathology, Microbiology & Immunology, UC Davis, CA and 3.

Department of Pathology, UTMB, Galveston, TX, USA

Species in the Lutzomyia longipalpis complex differ in the quantity of maxadilan, a

protein in the saliva. We have suggested that this difference may explain variation in the

pathology produced by Leishmania chagasi, for which these flies are the main vector.

Leishmania chagasi typically leads to visceral leishmaniasis in South America, but most

frequently causes a benign disease, known as “atypical cutaneous leishmaniasis” in

Central America.

To test our hypothesis in a setting close to natural infections, we compared parasite

dissemination in hamsters infected by bites of sand flies from two colonies: one high in

maxadilan (High-Max, Lapinha, Brazil) and one low in maxadilan (Low-Max, Brasilito,

Costa Rica).

Flies from the two colonies were infected side-by-side via membrane feeding on a

suspension of rabbit blood containing 2X106/ml L. chagasi amastigotes. At 12 days

post-infection, flies were allowed to feed on 4-week-old hamsters. Four months

following infective bites, animals were sacrificed and examined for infection.

A Quantative TaqMan PCR assay (QRT_PCR), together with microscopic analysis, was

used to assess tissue parasite loads. Of 8 animals bitten by the low-max flies, only two

(25%) showed amastigotes in the spleen tissues and one showed liver infection, with

average loads of 9 and 0.03 amastigotes/100 host cells in the spleen and liver,

respectively. In contrast, 6 of 7 (85.7%) animals bitten by the high-max flies showed

high tissue parasite loads, with average of 33.7 and 12.0 amastigotes/100 host cells in

the spleen and liver, respectively. The differences in parasite loads were statistically

significant (Mann-Whitney p values of 0.005 for spleen and 0.013 for liver). Histology

evaluations revealed a correlation between parasite loads and inflammatory foci.

This is the first report indicating the impact of vector’s genetic differences on pathology

of the parasite they transmit, and this novel finding will be discussed in relation to the

epidemiology of L. chagasi infections in South and Central America.

Work funded by NIH/NIAID, grant number: 5R01AI039540-10

OP-40

SAND FLY SALIVA, HOST IMMUNE RESPONSE, AND LEISHMANIASIS

IVA ROHOUSOVA, JAN DRAHOTA, JITKA HOSTOMSKA, JAN VOTYPKA,

and PETR VOLF

Department of Parasitology, Charles University, Vinicna 7, 128 44 Prague 2, Czech

Republic, e-mail: rohousova@seznam.cz

The immunomodulation activity of sand fly saliva plays an important role in the

establishment of Leishmania parasites within the vertebrate host during the first steps of

infection. However, the effect varies between naive and saliva-immunised hosts.

Therefore, we analysed and compared immune response between naive and sand fly-

exposed mice.

In naive mice, salivary gland homogenate (SGH) significantly suppressed both

spontaneous and concanavalin A-stimulated lymphocyte proliferation. In repeatedly

bitten mice, no significant differences were found between control and SGH treated

cells. In parallel experiments we compared immunomodulation activity of different sand

fly species. In naive hosts, all sand fly species tested modulated cell proliferation as

well as the cytokine production in a similar way. In repeatedly bitten hosts, we found

significant differences between cells challenged ex vivo with homologous or

heterologous SGH.

Additionally, we tested the effect of salivary enzyme hyaluronidase on

Leishmania establishment in the host. We showed previously that hyaluronidase is

present in all sand fly species tested. This enzyme plays an important role in blood meal

acquisition as it degrades hyaluronan and other components of extracellular matrix.

Moreover, breakdown products (hyaluronan fragments) are supposed to have

immunomodulatory properties. Thus, in the recent work we assessed the effect of

hyaluronidase coinoculation on the outcome of Leishmania major infection using a

mouse ear infection model. Mice coinoculated with L. major and hyaluronidase

developed bigger lesions than the controls inoculated with parasites only. Parasite

numbers in draining lymph nodes collected early after infection were similar in both

groups.

OP-41

PHLEBOTOMUS PAPATASI SALIVARY GLAND SEQUENCE VARIABILITY

AND IMPACT ON DEFINING VACCINE CANDIDATES

RAMI MUKBEL1, MARCELO RAMALHO-ORTIGAO

1, VINITA TRIPATHI

1,

VALDIR BALBINO1, ILIANO COUTINHO-ABREU

1, GWEN STAYBACK

1,

MARIHA WADSWORTH1, EMIL LESHO

5,

EMAD DIN YEHIA FAWAZ2,

SHAABAN S. EL-HOSSARY2, HANAFI HANAFI

2, DAVID HOEL

2, MAHMOUD

ABO-SHEHDA3, SHADEN KAMHAWI

4, GLENN WORTMANN

5, AND MARY

ANN MCDOWELL1.

1University of Notre Dame, Notre Dame, IN, USA

2Naval Medical Research Unit #3, Cairo, Egypt

3Jordan University of Science and Technology, Irbid, Jordan

4LMVR, NIAID, NIH, Rockville, MD, USA

5Walter Reed Army Medical Center, Washington DC, USA

Data suggest that the incorporation of salivary components in multi-component

vaccines may be a viable strategy for the development of anti-Leishmania vaccines. If

sand fly saliva as a vaccine component is to be realized, it is necessary to understand the

variability of salivary genes and human immune responses to such variability. Here we

investigated expression and amino acid sequence variability of salivary gland proteins

from field populations of P. papatasi sand flies from the Middle East. Salivary gland

cDNAs encoding secreted proteins were PCR amplified, sequenced and the results were

compiled using various bioinformatics tools. For each protein, predicted MHC class II

T-cell epitopes were obtained and compared to areas of amino acid sequence variability.

Our results indicate greater sequence variability than was previously suggested and we

have identified additional MHC class II T-cell epitopes. To explore the early events of

anti-saliva human immune responses monocyte derived dendritic cells and macrophages

were generated from naïve individuals and the effect of sand fly salivary gland

homogenate (SGH) on co-stimulatory molecule expression and cytokine secretion was

assessed. Blood samples also were collected from US soldiers deployed to Iraq and

compared to US soldiers that have never been deployed to P. papatasi endemic regions.

Salivary antigens recognized by the sera of exposed individuals were identified by

western blot analysis of P. papatasi SGH. Peripheral blood mononuclear cells were

assessed for anti-SGH proliferative capacity and cytokine secretion.

OP-42

ANTIBODIES ANTI-SALIVA OF LUTZOMYIA SPP. IN SMALL MAMMALS IN

AN ANDEAN LEISHMANIOSIS ENDEMIC AREA IN PERU.

J ENRIQUE PEREZ1, KATHERINE TORRES, MARGARITA ARANA

2,

OSWALDO RAMIREZ2, MIGUEL CAMPOS

2 & ELENA OGUSUKU

3

1 Instituto de Medicina Tropical Alexander von Humboldt, Universidad Peruana

Cayetano Heredia. kike@upch.edu.pe 2 Facultad de Ciencias y Filosofía, UPCH.

3 Dirección General de Salud Ambiental, Ministerio de Salud, Peru.

The animals of leishmaniosis endemic areas are exposed to sandfly bites and are source

of blood in the maintaining of the sandfly population.

OBJECTIVE: The objective of this study was to detect antibodies antisaliva of

Lutzomyia spp.in small mammals in Chaute (Huarochirí, Lima), at 2500 masl, where

Andean cutaneous leishmaniosis (uta) is endemic.

METHODS: Small mammals were captured using Sherman traps in Chaute. The

antibodies antisaliva of Lutzomyia spp. were detected through the ELISA technique

using salivary gland homogenates of Lutzomyia peruensis and L. verrucarum from

colony.

RESULTS: 28 small mammals were collected, 14 Mus musculus, 8 Phyllotis andium, 3

Oryzomys xantheolus, 3 Thylamis pallidior. The titres of anti-saliva antibodies against

L. verrucarum saliva were higher than those anainst L. peruensis saliva (p=0.007, in a

paired t-test). Analyzing the species, Thylamis pallidior p=0.008 had a significant result

in despite of its low number n=3. The antibodies against Lutzomyia verrucarum saliva

are more common than those against L. peruensis saliva in Chaute. The production

against L. peruensis saliva is the lesser.

CONCLUSIONS: The antibodies against L. verrucarum saliva are the most competent

and would control a Lutzomyia mediated Leishmania infection more effectively.

The antibodies against L. peruensis saliva are the less competents, and for this reason, a

Leishmania infection will success. Lutzomyia peruensis becomes more effective vector

of Leishmania because the antibodies against its saliva are less competent.

This study received financial support from Fondo Concursable, Vicerrectorado de

Investigación, UPCH. Directorate-General for Development Coperation, Belgian

Government, framework agreement 02.

OP-43

ANTIBODIES AGAINST SAND FLY SALIVA: USEFUL TOOL FOR

ANTIVECTORIAL CAMPAIGNS

JITKA HOSTOMSKA, IVA ROHOUSOVA, MICHAELA VLKOVA, VERA

VOLFOVA and PETR VOLF

Department of Parasitology, Charles University, Vinicna 7, 128 44 Prague 2, Czech

Republic, e-mail: volf@cesnet.cz

A repeated exposure to sand flies results in a specific antibody response in bitten

hosts. To find out whether anti-saliva antibodies could be a useful marker of exposure,

we studied the antibody response in dogs experimentally exposed to

Lutzomyia longipalpis females. The antibody response reflected the intensity of

exposure; a stronger IgG response to salivary antigens was observed in high-exposed

dogs. The difference in IgG production between high- and low-exposed dogs was

detectable throughout the study, i.e. more than 6 months after the last exposure. This

indicates that in endemic areas with seasonal fluctuations of sand fly populations, the

immune response to salivary antigens persists up to the following season.

Anti-saliva antibodies were found to be highly specific for different sand fly

species. Sera of mice, hamsters, and rabbits bitten by one sand fly species reacted

strongly with a homologous antigen and faint cross-reaction was observed only between

closely related species. Similarly, sera of humans bitten by Phlebotomus papatasi and

P. sergenti did not cross-react with L. longipalpis salivary antigens. Anti-saliva

antibodies could be therefore used as an efficient marker of exposure to sand fly

species.

Moreover, we demonstrated an important correlation between anti-saliva

antibody levels and the risk of pathogen transmission. Human sera were collected in an

endemic focus of Leishmania tropica where P. sergenti is the vector, while P. papatasi

is refractory to this Leishmania species. In comparison with healthy individuals from

the same place, people with active lesions developed a higher antibody response to

vector saliva (P. sergenti), while levels of antibodies against the non-vector species

(P. papatasi) were equal in both groups. These findings have proved that the antibody

response to vector saliva can generally serve as a marker of risk for various vector-

borne diseases.

SESSION VIII: Sandfly surveillance

OP-44

SAND FLY SURVEILLANCE WITHIN ACTIVE VISCERAL LEISHMANIASIS

FOCI IN TBILISI, GEORGIA

E. GIORGOBIANI 1, G. BABUADZE

1, N. DOLIDZE

1, G. CHANTURIA

1,

D. SACKS 2, P. LAWYER

2, S. KAMHAWI

2

1National Center for Disease Control, Republic of Georgia;

2National Institute of Allergy and Infectious Diseases, NIH, USA

A total of 1,414 visceral leishmaniasis (VL) cases was reported in Georgia from 1990 to

2007, resulting in an 18 fold increase from 10-12 cases per year reported in the early

90’s to 182 cases reported in 2007. A weakened surveillance system for VL in Georgia

during the past 15 years led to a resurgence of the disease and the majority of cases

(~60%) occurred within emerging foci in the capital city, Tbilisi. Since the identities of

the responsible parasite, animal reservoir and vector species remained unknown two

active VL foci in Tbilisi, Vera and Vake communities, were chosen for investigation.

Entomological surveys were conducted over 3 consecutive years (2006-2008) using

light traps and sticky-paper traps to assess the sand fly species diversity, abundance, and

to incriminate the vector(s) of VL in these foci. The sand fly season started at the end of

May, peaked in June-July and ended by late August. The captured sand flies were

identified using a collection of taxonomic keys. In August 2008, live female flies were

dissected and examined microscopically for natural parasite infections. Positive sand

flies were subjected to PCR analysis for identification of Leishmania parasites using

specific primers (Uni21/Lmj4). Five sand fly species were collected during the survey.

Phlebotomus kandelakii was the most abundant species (64%), followed by P. sergenti

(19%), P. balcanicus (10%), P. halepensis (6%), and P. wenyoni (1%). About 90% of

sand flies were collected by light traps vs. 10% captured by sticky-paper traps. Of 324

females dissected, Leishmania promastigotes were found microscopically in three

specimens (infection rate ≈1%), one P. balcanicus and two P. kandelakii. The parasites

were identified by PCR analysis as L. infantum and the PCR product matched those

amplified from isolates of local patients and dogs. Our results suggest that P. kandelakii

is the primary vector of L. infantum in Tbilisi. However, detection of naturally infected

P. balcanicus is critical evidence of more than one vector responsible for transmission

of L. infantum in this focus.

OP-45

SUPPORT OF FAR FORWARD DISEASE SURVEILLANCE

OPERATIONS WITH DEPLOYABLE, REAL-TIME VECTOR-BORNE

DISEASE AGENT ANALYTIC CAPABILITY

COL JAMES A. SWABY AND JAMES C. MCAVIN1

1. 59th Clinical Research Training Division, 59th Medical Wing/Air Force Surgeon

General for Modernization (MDW/SGR), Lackland AFB, Texas 78236-9908

The Vector Surveillance Analytic System (VSAS) is a portable, field-durable, field-

sustainable, real-time, arthropod-borne disease agent detection platform used to support

disease surveillance operations at far forward locations. The VSAS operates as a stand-

alone field surveillance activity or as an extension of other deployable assets. The field

utility of the VSAS is clearly proven in diverse operational applications and

environmental conditions. The VSAS provides deployable analytic capability for real-

time vector-borne disease risk assessment which is paramount in affecting time-critical

and focused disease preventative and control measures.

OP-46

SURVEILLANCE AND CONTROL OF SANDFLIES AS VECTORS OF

ZOONOTIC CL IN NORTHERN AFGHANISTAN

ANDREAS KRUEGER1, THOMAS MORWINSKY

2, HENRI DERSCHUM

3,

MOHAMMED AMIRIH4, WINFRIED MAASSEN

2, HANS-ULRICH

HOLTHERM2, GERHARD HEYL

2, JOACHIM SCHRADER

2, MICHAEL

FAULDE5

1. Bundeswehr-Department of Tropical Medicine, Bernhard Nocht Institute for

Tropical Medicine, Hamburg, Germany

2. Division of Preventive Medicine, Bundeswehr Medical Office, Munich,

Germany

3. Bundeswehr Institute for Microbiology, Munich, Germany

4. Balkh Province Leishmaniasis Center, Mazar-e Sharif, Afghanistan

5. Department of Medical Zoology, Central Institute of the Bundeswehr Medical

Service, Koblenz, Germany

When, in 2005, International Security Assistance Forces (ISAF) troops began to set up a

new camp at Mazar-e Sharif (MeS) airport in northern Afghanistan, about 200 cases of

zoonotic cutaneous leishmaniasis (ZCL) were recorded among them. This led to a quick

establishment of vector and rodent reservoir surveillance, which in turn revealed the

highest population density of the great gerbil (Rhombomys opimus) ever observed so

far. In addition, the primary vector of the causative agent of ZCL Leishmania major, i.e.

Phlebotomus papatasi, was recorded in high numbers. As a consequence, the new camp

was designed for prevention of both gerbil and sandfly habitats, resulting, together with

personal protection measures, in almost zero new infections since 2006/7.

Epidemiological analyses of the transmission dynamics showed a two-peaked seasonal

distribution of CL cases, which could be correlated with the anthroponotic form (L.

tropica) during winter-spring and the more prevalent zoonotic form (L. major) during

late summer-fall. Further entomological parameters are compared with more recent data

from another camp in a mountainous area near Feyzabad (with the presence of R.

opimus being questionable) and with published data from Kabul and Iraq. Whereas in

MeS almost 75% of captured sandflies belong to P. papatasi (and only 2% to secondary

vector species), the proportion of P. papatasi in Feyzabad in August 2007 was only 1%,

whilst 77% of all sandflies were non-mammalophilic Sergentomyia spp. The remaining

sandflies were mainly P. (Paraphlebotomus) alexandri.

OP-47

HOUSE AND ENTOMOLOGICAL RISK FACTORS FOR CUTANEOUS

LEISHMANIASIS IN THE SUBANDEAN AREA OF SOUTHWEST

COLOMBIA

RAUL HERNANDO PARDO1, CLIVE RICHARD DAVIES

2

1Universidad de La Salle, Bogotá, Colombia;

2London School of Hygiene & Tropical

Medicine, London, UK.

Introduction. Cuteneous leishmaniasis (CL) is the major leishmaniasis problem in

Colombia, with most cases located in the sub-andean region. Risk factors for CL,

including sandfly vectors, in this region are poorly understood.

Objective. To identify indoors, and around houses, risk factors for CL in the sub-

andean area of Colombia.

Methods. A house based cross sectional study was carried out in three sub-andean rural

areas within an epidemic region for CL in the department Huila, Colombia. Information

on cases and potential risk factors (house and surrounding habitats features and number

of potential hosts) for CL and sandfly suspected vectors was collected by

questionnaires. In addition, sandfly abundance data was recorded by indoors catches

with CDC light traps. Logistic regression analysis was used to test for CL risk factors

and to test for the relationship between sandfly abundance and CL prevalence.

Results. A total population of 1427 inhabitants was recorded in 271 sampled houses.

Total cumulative prevalence was 11.4% with significant higher prevalence in males

compared with females. Sandflies were present in 89% of the sampled houses with a

total of 7,659 caught sandflies. L. longiflocosa was the dominant species (93.5%)

followed by L. nuneztovari (2.1%). Risk factors for CL detected by multivariate

analysis were: village, altitude, gender, length of residence in the house, abundance of

L. longiflocosa (positively associated) and L. nuneztovari (negatively associated)

females.

Conclusions. The results point to L. longiflocosa as the most important vector of CL

within the study area and indicate a significant degree of transmission indoors.

Furthermore, the results provided the rational for testing interventions aiming to prevent

CL by reducing indoor exposure to L. longiflocosa bites.

Financial support. Main financial support for this study was provided by

COLCIENCIAS, Instituto Nacional de Salud, Pan American Health Organization and

Huila Health Service.

SESSION IX: Sandfly control

OP-48

AFPMB-DWFP SUPPORT FOR DEVISING NEW METHODS AND

MATERIALS FOR CONTROLLING PHLEBOTOMINE SAND FLIES

GRAHAM WHITE

DWFP Consultant, U.S. Armed Forces Pest Management Board, Forest Glen,

Maryland, U.S.A.

Since 2004, the AFPMB of the US-DoD has provided $5 million/year for research

projects to find and develop new insecticides and better application methods for

controlling disease vector mosquitoes, phlebotomines, and muscoid flies. Known as the

R&D program for Deployed War-Fighter Protection (DWFP), projects include

screening chemical libraries from academic and commercial partners, finding

insecticides with new modes of action, reformulating agricultural pesticides suitable for

controlling public health pests, design and improvement of spraying systems, field

evaluations of vector control tools and strategies. Based on fortifying collaboration with

USDA-ARS National Program for Veterinary Urban & Medical Entomology, the

DWFP program also awards grants of up to $250K for 3 years to competitive proposals

from academics, industry, military entomologists and other government agencies, not

restricted to USA. An overview will be given of multidisciplinary activities at 5 USDA

labs, plus 12 grants for competitive projects targeting phlebotomines.

OP-49

KNOWLEDGE OF SAND FLIES FOR PUBLIC AND DOMESTIC CONTROL

ACTIVITIES OF KALA-AZAR IN RURAL BIHAR.

NARENDRA KUMAR, N A SIDDIQUI, A RANJAN, R B VERMA, P. DAS.

Affiliation: Rajendra Memorial Research Institute of Medical Sciences (ICMR) Patna-

800 007 India

Introduction: Visceral leishmaniasis (VL) control is a global cause of concern. The

main aim of (VL) control programme is to reduce morbidity, mortality and transmission

of disease. Kala-azar is one of the most neglected diseases in the world, affecting

poorest segment of rural populations in Southern Asia, Eastern Africa and Brazil. This

disease has been one of the major public health problems in the state of Bihar, India for

many decades. . Presently, 28 out of 37 districts in Bihar are endemic at various levels.

More than 90% of all cases in India are reported from Bihar alone. The increasing case-

fatality rate observed during the past 10 years is a matter of serious concern among

health care providers and policy makers.

Objective: To identify the gaps in people’s knowledge/ awareness about sand flies and

control activities of Kala-azar in rural endemic areas of Bihar.

Methods: A house-hold based cross-sectional community survey was conducted. The

total numbers of households covered in the four villages were 450, with a total

population of 4012 individuals belonging to all age groups and both genders were

covered.

Result: The result showed that 95% respondents had heard about the disease up to some

extent, but surprisingly 99.1% respondents were neither aware about the vector of Kala-

azar, nor they had any idea about transmission of the disease. About 61 % had wrong

impression that mosquitoes were causing Kala-azar. Knowledge about breeding and

resting sites of vectors was quite satisfactory, as 20% reported cattle shed 16% crevices

in the household followed by 15% damp dark places. The attitude of respondents

towards vector control programme was poor, as 99% lost faith in the DDT spraying

because of ineffectiveness and no reduction in mosquito nuisance. Bed net was

considered the best protection method against sand fly or mosquito nuisance but the cost

was considered the major constraint in its use.

Conclusions: Proper health education programme in simple and local language along

with visual demonstration should be promoted to enhance the awareness and co-

operation at community level.

OP-50

PERSPECTIVES OF SAND FLY CONTROL IN THE JUDEAN DESERT

LAOR ORSHAN1

AND ORNA MATZNER2

1Laboratory of Entomology, Ministry of Health, Jerusalem

2Pest Control Division, Ministry of Environmental Protection.

The principal vector of Leishmania tropica in the Judean Desert is Phlebotomus

sergenti, which is considered an outdoor species in Israel. The local control methods

consist of spraying residual insecticides on house surfaces as a barrier against incoming

sand flies.

A sand fly control study was conducted in the town of Ma'ale Adummim in the Judean

Desert, 10km east of Jerusalem. The houses are built on the edges of slopes, separated

by uninhabited valleys. Supportive 3-15 m rockeries or walls separate the small

peripheral backyards from the desert slopes.

The goals of the study were (1) to determine the optimum spray locations, (2) to test the

efficacy of creating a treated barrier to reduce sand fly populations and (3) to determine

the residual efficacy of insecticides exposed to desert conditions in the summer months.

Sand flies were collected using modified CDC light traps baited with dry ice and placed

in fixed positions on the rockeries and in backyards. Residual insecticides consisting of

pirimiphos methyl EC (2%), microencapsulated and EC chlorpyrifos formulations (1%)

and cypermethrin EC (0.5%) were applied at a rate of 1 liter per 10 m2. Large scale

treatments were conducted by the local municipality. Small scale experiments were

conducted on 10x10 m2 of rockeries and cloth covered areas. In these experiments, we

also tested bifenthrin EC (0.5%).

Based on the numbers of sand flies and the low proportion of males collected in traps,

our results indicated that supportive rockeries and walls were suitable spraying sites for

the application of residual insecticides to control sand flies. Moreover, the application

of sugar solutions containing different food dyes to the sprayed area indicated that sand

flies travel from rockeries into yards.

None of the conventional insecticides tested reduced the number of sand flies collected

in backyards. On the sprayed surfaces, only bifenthrin reduced the number of sand flies

for one week. Chemical analysis and mortality in sand flies after forced contact with

treated surfaces showed that the lack of efficacy in the field was not a result of chemical

degradation of insecticides. Possible explanations and the significance of the results for

sand fly control by surface treatments and future perspectives are discussed.

OP-51

FEED-THROUGH CONTROL FOR LARVAL SAND FLIES USING HOST-

TARGETED INSECTICIDE.

GIDEON WASSERBERG1, DAVID MILLER

2, EDGAR RAWTON

1

1Walter Reed Army Institute of Research, Silver Spring, MD.

2Genesis Laboratories Inc. Wellington, CO.

Because Phlebotomine sand fly larvae are known to feed on the feces of rodents, this

behavior can be exploited as a potential route for sand fly control. Here, we describe

the results of a pilot study aimed to evaluate the feed-through effectiveness of the

insecticide Imidoclorpid on larval sand flies. Eight laboratory rats were exposed for 7

days to a diet containing the insecticide Imidoclorpid (100 and 250 ppm). Another 4 rats

were fed standard rat feed (control). Fecal pellets were collected 2 days and 4 days after

diet feeding. These 24 plus 6 additional samples containing standard sand fly larvae

feed were used for the sand fly larvae feeding assay. Fecal and standard feed samples

were crushed and pulverized and applied to different wells (3.5 cm diameter) containing

2-3 days old larvae of Phlebotomus papatasi (Israeli strain) or Lutzomyia longipalpis

(Jacobina strain). For both sand fly species a typical dose-response was observed with

strongest lethal effect for the 250 ppm samples, less for the 100 ppm samples, and no

effect for the control samples. For both species, at either dose, samples from pellets

collected at day 4 had a stronger effect than pellets collected at day 2. At any dose, P.

papatasi larvae are more sensitive to the treatments compared with L. longipalpis.

These results support the potential efficacy of feed-through methods using Imidoclorpid

as a potential method for sand fly control.

OP-52

EVALUATION OF LONG LASTING INSECTICIDAL NETS (LLINS) ON THE

FEEDING BEHAVIOR OF SAND FLIES (DIPTERA: PSYCHODIDAE) IN THE

LABORATORY.

GABRIELA E. ZOLLNER, ROXANNE FLORES, EDGAR ROWTON,

RUSSELL E. COLEMAN.

Division of Entomology, Walter Reed Army Institute of Research (WRAIR), Silver

Spring, MD, USA.

Leishmaniasis is a disease transmitted by the bite of an infected phlebotomine sand fly.

In the US military, sand fly vector control emphasizes the use of Personal Protective

Measures (i.e. DEET-based repellents, permethrin-treated uniforms and bed nets). The

standard military bed net has a very fine mesh (920 holes per sq. inch) and provides a

physical barrier against sand flies, but it requires treatment by the user and is

claustrophobic to use in hot climates. We have teamed up with manufacturers of LLINs

to develop a breathable netting material that is effective against sand flies. In a series of

laboratory tunnel assays, we evaluated three LLINs (Interceptor(r), Olyset(r),

PermaNet(r)) in different mesh sizes (156-625 holes/sq. inch) and impregnated with

different pyrethroid insecticides (α-cypermethrin, permethrin, deltamethrin) for their

efficacy in preventing Phlebotomus papatasi sand flies from feeding on mice. Twenty

flies were released into one end of a tunnel chamber and had to pass through treated

netting to feed on a mouse located at the other end of the tunnel. Assays with treated

netting were paired with two controls consisting of tunnels containing untreated netting

or no netting. Deltamethrin LLIN provide the highest initial contact irritancy and

subsequent knockdown, but α-cypermethrin LLIN caused higher feeding inhibition.

Surprisingly, of a total 60 sand flies that were able to squeeze through α-cypermethrin-

or deltamethrin-treated netting with the finest mesh (625 holes/sq. inch) and feed on the

mouse, approx. 30% of the flies still survived 24 h later. This study raises the question

of how sand flies pass through pyrethroid-treated netting materials with a fine mesh but

do not pick up a lethal dose of insecticide.

OP-53

WHAT DO SAND-FLIES WANT? UNVEILING SECRETS OF HOST AND

PLANT ATTRACTION.

EDGAR RAWTON1, PHILIPP KIRSCH

2, GIDEON WASSERBERG

1

1Walter Reed Army Institute of Research, Silver Spring, MD.

2APTIVE Inc., Portland, OR

Chemical cues are necessary for sand-fly survival. They are used to locate and feed on

mammalian hosts or plant sugars. Sand-fly sugar feeding behavior is critical to adult

survival and as well as being necessary for Leishmania development and transmission.

The goal of this research was to study the topic of chemical cues that attractant sand-

flies. Our specific goal was to screen a wide range of putative plant and host attractants

in order to identify the biologically active compounds comprising the strongest sand-fly

attractants. Here we present results from an experiment in which we used olfactometer

experiments. The olfactometer is a cylindrical apparatus (46.5 x 10 cm) made of

Plexiglas and composed of 3 chambers 15.5 cm in length. Air flows through a filter and

then splits to each side of the olfactometer. The chemical is placed in one of the outer

chambers and the sand-flies (20 per experiment) are introduced into the central

chamber. The vacuum is attached to pull the air through the system and the system runs

for 1 minute before the perforated doors to each outer chamber are opened. The doors

are left open and the test is run for 15 minutes to allow flies to distribute themselves

between the three chambers (treatment, center, or control). The doors are then closed

and flies in each chamber counted. Here, we present the results of experiments with

Phlebotomus duboscqi Kenya strain (51 compounds) and P. papatasi Israeli and

Turkish strains (5 and 7 compounds, respectively). For P. duboscqi only 4 compounds

(plant-derived sugars) were found to have a statistically significant (P<0.05) attracting

effect and another 5 with a marginally significant attracting effect. Two of the plant-

derived compounds found to be attractive to P. duboscqi were also attractive for P.

papatasi Turkish strain. For P. papatasi Israeli strain, only one plant-derived compound

was found to have a significant attractive effect. Further screening experiments are

being done. Results of this study will allow us to better understand the chemical cues

the drives sand-fly’s attraction to food or host. This study also has direct applicative

implications. It would allow us to optimize sand-fly attractant blends that could be used

for targeted surveillance and for the production of attract-and-kill traps that could

potentially be used as an area-wide sand-fly control mechanism.

SESSION II: Ecology, Distribution, and Population Dynamics of Sandflies

(Addition).

OP-54

SANDFLY FAUNA OF CENTRO DE INVESTIGACION Y CAPACITACION

RIO LOS AMIGOS (CICRA), MADRE DE DIOS, PERU.

PEREZ1, J. ENRIQUE; DIANA RADO

2, WILFREDO QUISPE

2 & ELENA

OGUSUKU3

1 Instituto de Medicina Tropical Alexander von Humboldt Universidad Peruana

Cayetano Heredia. kike@upch.edu.pe 2

Universidad Nacional de San Antonio Abad del Cusco

3 Direccion General de Salud Ambiental, Ministerio de Salud, Peru.

INTRODUCTION: The tropical forest areas located Eastern of the Andean Cordillera in

Peru shows a great faunal diversity, the diversity of Lutzomyia sandflies (Diptera:

Psychodidae) is very high in the Department of Madre de Dios, some areas like the

Tambopata Reserve have 35 species.

OBJECTIVE: To determine the composition of the sandfly fauna of Centro de

Investigacion y Capacitacion Rio Los Amigos (CICRA), Madre de Dios, Perú.

METHODS: Four trips were made to collect sandflies during 2007. The collecting

methods used were: Shannon trap with protected human bait from 18:00 to 22:00, and

CDC light traps operated from 18:00 to 06:00. The traps were placed in different types

of vegetation.

RESULTS & DISCUSSION: Twenty five species were identified for CICRA, 5 species

are new records for Perú, these species are: Lutzomyia andersoni, L. corossoniensis, L.

gantieri, L. monstruosa, and L. recurva. Four species remain to be identified, these are:

L. near howardi, L. near olmeca bicolor, and L. sp. 1 and L. sp 2, from the Subgenus

Trichophoromyia.

The “Brazil nut” or “castaña” forest gave the highest sandfly densities, the most

abundant species in this was Lutzomyia paraensis (94 %), specially in the Shannon trap

with protected human bait collections, this species is very highly antropophilic. This

finding suggest an idea of the high exposure to Lutzomyia paraensis bites the Brazil nut

collectors are in the season of this fruit.

This study received financial support from Asociacion para la Conservacion de la

Cuenca Amazonica (ACCA).

.

POSTER PRESENTATIONS

POSTER SESSION I

I: Systematics and Taxonomy of Phlebotomine Sandflies

P-01

A POPULATION OF LUTZOMYIA LONGIPALPIS S.L. (LUTZ & NEIVA, 1912)

(DIPTERA: PSYCHODIDAE: PHLEBOTOMINAE) WITHOUT TERGAL SPOT

ANDREY JOSÉ DE ANDRADE1, FELIPE FONSECA DO CARMO

1, MATEUS

RAMOS DE ANDRADE1, ÁLVARO EDUARDO EIRAS

1, REGINALDO

PEÇANHA BRAZIL2 & JOSÉ DILERMANDO ANDRADE-FILHO

3

1Departamento de Parasitologia, Universidade Federal de Minas Gerais, Belo

Horizonte, MG, Brasil - bioandrey@gmail.com 2Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, Rio de Janeiro, RJ, Brasil

3Centro de Pesquisas René Rachou, Fundação Instituto Oswaldo Cruz, Belo Horizonte,

MG, Brasil

Lutzomyia longipalpis s.l. (Lutz & Neiva, 1912) play an important role in the

transmission of Leishmania (Leishmania) infantum Nicole, 1908, in the Central and

South America. There are some morphological variations in male abdomens of L.

longipalpis: (a) with a single pair of tergal patches on tergite IV; (b) other with two

pairs on tergite III and IV. There are, however, in natural populations, intermediate

forms with tergal spot on segment III showing some variation. In this report, we show a

new morphotype in males of L. longipalpis. Studies have been carried in Chapada

Gaúcha, North of the State of Minas Gerais (Brazil). Phlebotomine sandflies were

caught by CDC light traps placed in a chicken pen within a peridomiciliary

environment, between 18:00h and 08:00h during two weeks. We observe three

morphological variations in tergal spots on L. longipalpis s.l. males of Chapada Gaúcha:

(a) single spot on segment IV; (b) two spots on segments III and IV; (c) neither tergal

spot on the segments. In Brazil sympatric populations of L. longipalpis have been found

in several regions. The present study shows for the first record of a triple sympatry of

males of L. longipalpis in this country. The morphotypes (c) present intumescences on

the III and IV tergites, but not pale spots. This variation will be further studied in

relation to the morphological aspects of the glands and it sexual pheromone.

Grants: CNPq, Fiocruz, Faperj, FINEP, SEBRAE

P-02

DESCRIPTION OF A NEW PHLEBOTOMINE SPECIES (DIPTERA:

PSYCHODIDAE) FROM THE STATE OF ESPÍRITO SANTO, SOUTHEAST

BRAZIL

JOSÉ DILERMANDO ANDRADE FILHO1, ISRAEL DE SOUZA PINTO

2,

CLAUDINEY BIRAL DOS SANTOS2 & GUSTAVO MAYR DE LIMA

CARVALHO1

1 - Laboratório de Leishmanioses, Centro de Pesquisas René Rachou/Fiocruz

2 - Unidade de Medicina Tropical, Universidade Federal do Espírito Santo

Phlebotomine sandflies presents great diversity of species, mainly in Brazil,

where there are known around 300 species. This is due in part to the large number of

biomes and ecotypes within the national territory, these environments able to maintain

several kind of biodiversity. This work describes a new species of sand fly found in the

Brazilian State of Espírito Santo, in the “Reserva Biológica de Duas Bocas”,

municipality of Cariacica. Sandflies were mounted in Berlese liquid and measured with

a binocular Olympus CH-2 microscope with the aid of a micrometer objective and the

drawings were realized with a camera lucida. The type-material is composed of three

males and four females. The morphological characters of the new species permit to

include it in the Evandromyia genus. Others aspects, such as, spemathecae with

superficial striations and the common duct longer than the genital fork permit include

the new species in Evandromyia s. str., series rupicola. This seies is composed by

Evandromyia rupicola (Martins Godoy & Silva, 1962), Evandromyia correalimai

(Martins, Coutinho & Luz, 1970), and Evandromyia gaucha Andrade Filho, Souza &

Falcão, 2007. The new species resemble E. rupicola and can be separated with security

by the aspects of the spermatheca in the females and the males can be separated by the

length of the genital filament, which are longer in the new species.

Financial Support: Fiocruz and Fapemig

Financial Support: FAPEMIG

P-03

DESCRIPTION OF A NEW SPECIES OF PHLEBOTOMINE SAND FLY IN

THE AMAZON BASIN OF PERU (DIPTERA: PSYCHODIDAE)

R. FERNÁNDEZ1, V. LÓPEZ

1, E. REQUENA

2AND J. STANCIL

1

1 United States Naval Medical Research Center Detachment, Lima, Peru.

2 Universidad Nacional de la Amazonia Peruana, Iquitos, Peru

The Lutzomyia subgenus Trichophoromyia includes numerous species that are widely

distributed in the Amazon basin. They are generally zoophilic, feeding on non-human

primates and other small mammals. The Trichophoromyia subgenus was first

documented in the Peruvian Amazon with the description of Lutzomyia loretonensis

(Llanos 1964) collected in the Loreto department of Peru. In 1994, Young & Duncan

described three new species from the Peruvian Amazon, L. clitella, L. nemorosa and L.

sinuosa, for a total of 9 species reported nationally. Additional studies in Peru have

accumulated a total of 13 species in the Trichophoromyia subgenera. In this report we

document the presence of new species records, L. cellulana, L. saltuosa, L. brachipyga,

L. velascoi collected in Peru. Additionally, in 2003, personnel from the University

Nacional de la Amazonia Peruana and the Naval Medical Research Center Detachment

conducting sand fly collections in the Loreto department of Peru (utilizing CO2-baited

CDC light traps and human-baited Shannon traps) discovered a previously undescribed

species of the Trichophoromyia subgenera, which we have named Lutzomyia nautaensis

sp.n. Differentiation of L. nautaensis sp.n from other Trichophoromyia is facilitated,

however, by the presence of two bristle tufts in the basistyle, one of them basal and

compact and the second, apical and diffuse and the paramere which is simple and

typical of the species.

P-04

MORPHOLOGY AND MORPHOMETRY STUDIES OF LUTZOMYIA

(LUTZOMYIA) LONGIPALPIS (DIPTERA:PSYCHODIDAE) POPULATIONS,

VECTOR OF VISCERAL LEISHMANIASIS IN BRAZIL

MARGARETE M S AFONSO1, MARA G TAVARES

2, JOSÉ C MIRANDA

3,

LINDEMBERGH C SOUSA4 & ELIZABETH F RANGEL

1

1Lab. Transmissores Leishmanioses, Instituto Oswaldo Cruz-IOC/FIOCRUZ,Av. Brasil,

4365,Rio de Janeiro,RJ,Brasil 2Depto. Biologia Geral, UFV;

3Lab. Imunoparasitologia,

CPqGM/FIOCRUZ; 4Lab. Entomologia Médica Dr. Thomaz Correa Aragão, SESAU-

CE, Brasil

Lutzomyia (Lutzomyia) longipalpis is considered a complex of species, although in

Brazil, this hypothesis still generates certain disagreement over the real taxonomic

status of this vector. The aim of this study is to compare the morphology and

morphometry of male sand flies, evaluating the degree of polymorphism within and

among Brazilian populations from Teresina/PI, Jequié/BA, Sobral and Massapê/CE (the

last two were considered four populations, according to different patterns of tergal

abdominal spots: S1P, S2P, M1P, M2P) considering 21 characters, analyzed by

ANOVA and SNK tests. Morphological data revealed individuals with one and two

pairs of abdominal spots in all populations. The most frequent palpal formula was

1.2.4.3.5. Morphometric analysis by SNK test showed unions, disjunctions and

intersections between the six populations. Based on the analyses of six characters it

could be observed that Jequié and Teresina populations produced an individual set from

those formed by the Ceará populations. In population of Jequié, when compared to

population of Ceará, it can be observed that 79% to 95% of the characters revealed

significant differences. By ANOVA, at the crossroads between the populations of

Ceará, few differences were detected. The dendogram created by UPGMA method

revealed a split between the Ceará and other populations. The males from Sobral, S1P

versus S2P, were clustered in two isolated branches of the tree. Bootstrap values

reinforced that populations from Ceará and Teresina/Jequié are in fact isolated, with a

confidence level of 100%. The data gathered by morphometry within and among

populations suggests L. (L.) longipalpis as a remarkable polymorphic species. However,

considering the sympatric speciation hypothesis, the morphometric evidences suggest in

Sobral the existence of two populations in process of speciation.

Financial Support:FIOCRUZ, DECIT-CNPq/2006

P-05

DESCRIPTION OF A NEW PHLEBOTOMINE SPECIES FROM A CAVE IN

TOCANTINS STATE, BRAZIL (DIPTERA: PSYCHODIDAE:

PHLEBOTOMINAE).

GUSTAVO MAYR DE LIMA CARVALHO, ALDA LIMA FALCÃO,

CRISTIANI DE CASTILHO SANGUINETTE, JOSÉ DILERMANDO

ANDRADE FILHO

Laboratório de Leishmanioses - Centro de Pesquisas René Rachou - Fiocruz

As inhabitants of forested areas, caves, and anthropic environments, the phlebotomines

call for special attention, due to the capacity of some species to transmit agents, such as

trypanosomatids, bacteria, and viruses, to vertebrates, as well as to cause bother by their

painful bites, which may produce allergic manifestations. The lack of information on

the presence of sandflies in caves and on its behaviour in this ecotype arouses the

curiosity of researchers a long time ago. The present paper deals a new specie of sand

fly captured in year of 1965 in a cave located in Arraias municipality, in the

southeastern state of Tocantins, to 413 km away from Palmas, capital of the state. The

cave was known as the Lapa das Cabeceiras, however, today is called Grutas da Lapa, a

complex of four large rooms, about 40 meters long and 20 meters high. The specimens

of new specie were deposited in the Collection of Sandflies of the “Centro de

Referência Nacional e Internacional para Flebotomíneos” of the Instituto René

Rachou/Fiocruz. A new species is described based in six females and eight males. The

morphological characters of the new species permit included in the Martinsmyia genus,

alphabetica group, being close to M. brisolai (Le Pont & Desjeux, 1987), M. oliveirai

(Martins, Silva & Falcão, 1970) and M. minasensis (Mangabeira, 1942). Both male and

female, has proboscis larger than those found in the others species. In addition, the

males can be separated using the length of the filaments genitals, disposition of the

spines of gonostyle and shape of paramere and the females, through the length of

common duct of the spermathecae, width of individual ducts and by elements of

cibarium.

Financial Support: IRR/FIOCRUZ, FAPEMIG

P-06

DESCRIPTION OF A NEW PHLEBOTOMINE SPECIES OF THE

CORTELEZZII COMPLEX, FROM CAVES OF THE MINAS GERAIS STATE,

BRAZIL (DIPTERA: PSYCHODIDAE: PHLEBOTOMINAE).

GUSTAVO MAYR DE LIMA CARVALHO1, REGINALDO PEÇANHA

BRAZIL2, CRISTIANI DE CASTILHO SANGUINETTE

1, JOSÉ DILERMANDO

ANDRADE FILHO1

1- Laboratório de Leishmanioses - Instituto René Rachou - Fiocruz

2- Laboratório deBioquímica e Fisiologia de Insetos - Instituto Oswaldo Cruz

Sandflies are responsible for the transmission of the protozoa of the genus Leishmania

among their vertebrate host and the study of this group of insects are of importance for

attempt in the control of leishmaniases. Phlebotomines are distributed throughout the

world and there are now about 500 known species in the Americas. The cave fauna of

the Brazil is poorly documented, and among the insects that live or frequent caves and

their adjacent environments, the phlebotomines call for special attention because several

species are vectors of diseases The growing tourism involving search for natural

attractions, as cave exploration, demands a better knowledge of the threats to health that

people may face. A new species from Minas Gerais is described based in females and

males of sandflies collected in caves of the municipality of Lassance. The

morphological characters of the new species permit included in the Evandromyia genus,

in cortelezzii complex. This complex consists of three species: Lu. corumbaensis

(Galati, Nunes, Oshiro & Rego, 1989), Lu. cortelezzii (Brethes, 1923) and Lu. sallesi

(Galvão & Coutinho, 1940). The new specie can be separate of the others the cortelezzii

complex through morphological characters. The males are next to Lu. cortelezzii and

Lu. corumbaensis, being separated through characteristics of paramere, lateral lobe and

tuft of the gonocoxite. The separation of the females with the others of the complex is

more confused, therefore, can be identified throught the characteristics of the

spermathecae and its ducts, beyond characteristics of the cibarium. The new species is

sympatric with Lu. sallesi in the local of capture.

Financial Support: IRR/FIOCRUZ, FAPERJ, FAPEMIG

P-07

STUDIES OF SOME LUTZOMYIA (NYSSOMYIA) WHITMANI S.L. BRAZILIAN

POPULATIONS, IMPORTANT VECTOR OF AMERICAN CUTANEOS

LEISHMANIASIS IN BRAZIL: CONTRIBUTION TO THE DISCUSSION ON

THE TAXONOMY IDENTITY.

PINTO¹, OCTAVIO -, DIAMAR COSTASILVA¹-DANIEL MOTTA

FERNANDES DA SILVA FILHO², ELIZABETH FERREIRA RANGEL¹

¹Lab. Transmissores de Leishmanioses, ²Lab. Epidemiologia Molecular de Doenças

Infecciosas - Instituto Oswaldo Cruz – FIOCRUZ - Av. Brasil, 4365 - Manguinhos, Rio

de Janeiro,RJ, CEP: 21040-360,Brasil

The sand fly Lutzomyia (Nyssomyia) whitmani s.l is incriminated as vector of

American Cutaneous Leishmaniasis (ACL), associate with two parasites: Leishmania

(Viannia) braziliensis (Northeast, Southeast, South and Central Brazil) and L. (V.) shawi

(North Brazil). However, beyond the proven ability to transmit two parasites,

differences in the behavior among distinct geographical populations have suggested that

this sand fly species would be a complex of sibling species. The exact identification of

these populations is important for epidemiological studies of the ACL, allowing the

better knowledgement of the transmission cycles, considering that this sandlfy species

occurs in the great majority of the Brazilian States. In this study was assessed the

genetic variability of L. (N.) whitmani s.l., from Buriticupu (MA), transmission area of

L. (V) shawi and L. (V) braziliensis, Ilhéus (BA), type-locality, with transmission of L.

(V) braziliensis and Paragominas (PA), transmission region of L. (V) shawi, through the

technique of RAPD-PCR (Random Amplified Polymorphic DNA – Polymerase Chain

Reaction). The DNA of sand flies from each locality was extracted and amplified.

Among the six primers tested, the first P4 (5'-AAGAGCCCGT-3 ') proved to be very

polymorphic, identifying one band of 200bp in some individuals from Buriticupu and in

all individuals from Paragominas; this band is absent in all individuals from Ilhéus and

in some from Buriticupu. These data suggest the occurrence of two distinct populations

in Buriticupu in sympatry and related to the transmission of two different Leishmania

species. Supported by: Fiocruz and CNPq-DECIT/2006

P-08

APPLICATION OF THE CYTOCHROME OXIDASE (CO1)

MITOCHONDRIAL SEQUENCE TO THE SYSTEMATICS OF THE SAND

FLY SUBGENUS PINTOMYIA (PIFANOMYIA)

LEE W. COHNSTAEDT1, LORENZA BEATI

2, ABRAHAM G. CACERES

3,

CHRISTINA FERRO4,AND LEONARD E. MUNSTERMANN

1

1 Yale University School of Public Health, New Haven, Connecticut, USA

2 Georgia Southern University, Statesboro, Georgia, USA

3 Universidad Nacional Mayor de San Marcos, Lima Peru

4 Instituto Nacional de Salud, Bogota, Colombia

Introduction. In the Andes mountains, several sand fly species of the Pintomyia

(Pifanomyia) subgenus (sensu Galati) are known vectors of American cutaneous

leishmaniasis. Approximately 40 species have been included is this taxon. Species

within the 5 taxonomic series of the subgenus—evansi, monticola, serrana, townsendi,

and verrucarum--are often difficult to identify by morphology and have not been fully

compared by molecular sequences or related by phylogenetic analysis.

Objective. A standard mitochondrial gene sequence was used to distinguish the

Pintomyia species at the series and the species level, as well as to indicate the

phylogenetic relationships among them.

Methods. A 667 base pair fragment of mitochondrial cytochrome oxidase-1 (CO1) was

used first to document the intraspecific variability among 56 specimens of Pintomyia

verrucarum collected from 8 localities across its range in Peru. Then, interspecific

variation was compared among an additional 10 species representing 4 of the Pintomyia

(Pifanomyia) series (verrucarum--2 species, evansi--1 species, townsendi--6 species,

and serrana--2 species) as regrouped by Galati.

Results. The Kimura two parameter (K2P) distances for intraspecific variation

averaged 0.5% and 2.4% for the verrucarum and townsendi series. Intraseries

divergence averaged 14.9%, 4.2% and 5.2% for the verrucarum + evansi, serrana and

townsendi series, respectively. A maximum likelihood phylogenetic analysis provided

significant support for detecting the geographic localities of P. verrucarum species

populations, as well as assigning species level identifications for 6 species and two

species pairs. In the townsendi series, the 2 indistinguishable pairs were P.

longiflocosa—P. sauroida and P. torvida—P. quasitownsendi. The phylogenetic

relationships among the 4 morphologically distinct sand fly series was established

without ambiguity.

Conclusions. The COI sequence clearly distinguished the series in the subgenus

Pifanomyia as well as most of the species within series and provided well-

supportedphylogenetic relationships among them. However for 2 closely related

species pairs in the series townsendi, the COI sequence was not sufficiently senstive.

Financial Support. United States National Institutes of Health grants U19 AI065866,

R01 AI056254 and NIH training grant PTG-2T32-AI07404.

P-09

UTILITY OF THE TERMINAL DOMAIN CARBOXILE OF CITOCHROME

OXIDASE I, LIKE BAR CODE IN THE TAXONOMIC DETERMINATION OF

LUTZOMYIA SPECIES. LEISHMANIOSIS VECTORS IN COLOMBIA

RAFAEL J. VIVERO G1, MARÍA A. CONTRERAS

1, CAROLINA TORRES G

1,

ANDRÉS LÓPEZ RUBIO2, SANDRA URIBE SOTO

2 , IVÁN D. VÉLEZ

1 .

1 Research Associate, Program for the Study and Control of Tropical Diseases -

PECET, University of Antioquia. SIU- 62 #52-59 laboratorio 632.

pecet@siu.udea.edu.co: rajovigo2001@yahoo.com, Medellín, Colombia. 2. Molecular

Systematics Research Group – National University

Introduction: The taxonomic determination of the species of the genus Lutzomyia

Franca 1924, found in endemic leishmaniasis transmission areas is an essential tool for

understanding the epidemiology of this antropozoonosis. Taxonomic identification

based on morphological characters is very subjective and in some cases having with the

existence of plesiomorphic within a species does not allow obtain a precise and fast

identification. These mitochondrial sequences are included in the Bar Coding Initiative,

that considers COI as the target sequence of the barcode to identify Lutzomyia species.

Objective: The purpose of this study was to evaluate the polymorphism of the terminal

domain carboxile of Citochrome Oxidase I at intra-specific and inter-specific levels in

some collected species of Lutzomyia spp.

Methods: Specimens were obtained from different regions of Colombia, according to

the standardized methodology of the Medical Entomology Laboratory of the PECET.

Some species that were difficult to identify based on morphological characters were

used for this study, these included: Lu longipalpis, L. trapidoi, L. betini, L. runoides, L.

antunesi, L. panamensis among others. The DNA was extracted and a 680 nt product

was amplified for PCR. The genetic analyses were conducted in order to determine

nucleotidic and aminoacidic polymorphism and entropy grade as a signal of the possible

diagnostic sites species–specific.

Results and Conclusions: Several haplotypes of Lu. Longipalpis were determined,

supported by their nucleotide divergence and genetic distance also correlated with the

geographical precedence. Species such as L. panamensis y L. antunesi showed a close

phylogenetic relationship. L. runoides had a high divergence in terms of nucletidic

polymorphism and genetic distance which is also related to the presence of ancestral

morphological patterns. For all species of the genus Lutzomyia analized, the taxonomic

identification given was consistent with molecular characteristics registered using the

carboxyl terminal domain of the gene Cytochrome oxidase I.

P-10

MOLECULAR VARIATION IN THE PARALYTIC GENE IN TWO SYMPATRIC

SIBLING SPECIES OF THE LUTZOMYIA LONGIPALPIS COMPLEX.

R. M. M. A. LINS1, N. A. SOUZA

2 AND A. A. PEIXOTO

1

1 Laboratório de Biologia Molecular de Insetos, IOC, Fundação Oswaldo Cruz, Rio de

Janeiro, Brazil. 2 Laboratório de Transmissores de Leishmanioses, IOC, Fundação

Oswaldo Cruz, Rio de Janeiro, Brazil.

The paralytic (para) gene from Drosophila encodes a voltage-dependent sodium

channel associated with insecticide resistance and male courtship-song production. We

isolated a ~380 bp homologous fragment of the para gene from Lutzomyia longipalpis

s.l. and used it to study the molecular variation and divergence between two sympatric

siblings of the Longipalpis complex from Sobral, Ceará State, Brazil. In this locality

males of these two sibling species can be differentiated by the presence of one (1S) or

two (2S) pairs of abdominal spots. At the present work we analysed 27 sequences of

Sobral 1S and 22 of Sobral 2S males. The results revealed para as the first molecular

marker presenting fixed differences between these two species. We also observed two

low frequency amino-acid changes in an otherwise conserved region of the channel,

suggesting the possibility that these mutations might be associated with incipient

insecticide resistance in this vector.

Financial Support: Howard Hughes Medical Institute, CNPq and Fiocruz-PAPES IV.

P-11

DESCRIPTION OF A NEW SPECIES OF PHLEBOTOMINE SAND FLY,

BELONGING TO THE GENUS EVANDROMYIA (DIPTERA: PSYCHODIDAE:

PHLEBOTOMINAE) OF RONDONIA STATE, BRAZIL

CRISTIANI DE CASTILHO SANGUINETTE1, FÁTIMA DOS SANTOS

2,

GUSTAVO MAYR DE LIMA CARVALHO1, JOSÉ DILERMANDO ANDRADE

FILHO1

1- Laboratório de Leishmanioses - Centro de Pesquisas René Rachou - Fiocruz

2- Fundação Nacional de Saúde, Porto Velho - RO

Phlebotominae are diptera of the family Psychodidae, responsible for the transmission

of leishmaniases among animals, including man. In Brazil, the disease causes serious

damage to public health and is of great importance to the study of epidemiological and

ecological aspects of its vectors. The “Parque Natural Municipal de Porto Velho”,

Rondônia, popularly known as “Parque Ecológico”, has a total area of 390.8 hectares

and is located at the north of the capital, about seven kilometers of the urban perimeter.

In the preserved forest, predominates the Ombrophilous Open Forest with Palm Trees.

The relief is wavy and uneven, presenting narrow valleys forming sinuous alluvial

areas. The soil is of Yellow Latosol type covered with about 20cm thick of organic

matter, forming a coverage of dark soil. The fauna is represented especially by birds,

rodents, bats, ophidians, and other animals of small size. In 2004, specimens of female

phlebotomines, different from species known until the moment, were captured with

CDC light traps at the “Parque Ecológico” of Porto Velho. The new species from

Rondônia is described based in three females of sandflies collected in the park. Under

optical microscope (Olympus CH-2) adapted with micrometer objective (Olympus), the

specimens have been measured and drawn with help of a clear chamber. The presence

of ventro-cervical sensillae, setae on anterior katepisternum margin and papilla on the

flagellomeres AIII, AV, and AXIII, permit to include the new species in the genus

Evandromyia. The vertical teeth are laterally displayed and the spermathecae is apple-

like shaped which would make it similar to Aldamyia subgenera. Females of the new

species can be separated from other phlebotomines of the genus Evandromyia, such as

Evandromyia lenti, Evandromyia carmelinoi, Evandromyia walkeri and Evandromyia

williamsi, mainly by the size and shape of the spermathecae and by the common duct,

which is minor than in other species.

Financial Support: FNS/RO, FAPEMIG

P-12

GENETIC VARIABILITY IN LUTZOMYIA UMBRATILIS (DIPTERA:

PSYCHODIDAE) POPULATIONS OF CENTRAL AMAZON

JUSTINIANO, SCB1, MAIA, JF

2 ; SANTOS, JMM

2.

1

Centro Universitário Nilton Lins, Manaus, Amazonas, Brasil 2nstituto Nacional de Pesquisas da Amazônia, Coordenação de Pesquisas em Ciências

da Saúde, Manaus, Amazonas, Brasil. justiniano@niltonlins.br

Lutzomyia umbratilis is the main vector of cutaneous leishmaniasis due to Leishmania

guyanensis in northern South America. It has been found naturally infected with this

species of Leishmania only east of the Rio Negro and north of the Rio Amazonas.

However, populations of this sand fly species are also present in areas south of the

Amazon river system, which may act as a geographical barrier to the Leishmania

guyanensis. Considering the importance of L. umbratilis as the main vector of

tegumentar leishmaniasis in the Amazon, we analyzed four populations of the species

from the Negro River based on ten isoenzymatic loci to detect their intra- and

interpopulation genetic variability and differentiation. Adult phlebotomines were

collected in four terra firme (non-flooded) primary and secondary forest regions of the

Amazon State (Manacapuru, Novo Airão, Manaus and Pitinga).It was used two types of

fractionation support: partially hydrolyzed amide at 12% and amide-agarose (2% and

1%, respectively). The data were analyzed using the statistical software BIOSYS and

ARLEQUIN. The four populations presented variation in six (PGI, HK, IDH, MDH1,

-GPDH, and PGM) out of the ten loci analyzed, being PGI and PGM the most

polymorphic, with four alleles in most of the populations analyzed. Considering the data

obtained for the four populations of L. umbratilis, it presented high genetic variability

(P = 60%; Ho = 0.070 0.036 - 0.103 0.051 and He = 0.084 0.033 - 0.106 0, 053)

for this marker. The analyzed Manaus population presented the largest mean number of

alleles per locus (2.3 0.4) and the largest heterozygosity (Ho = 0.103 0.051;

He = 0.106 0.053). while, the Pitinga and Manacapuru populations presented the lowest

heterozygosity (Ho = 0.70 0.036 and He = 0.084 0.033, respectively). The analysis

of these data as a whole shows that the L. umbratilis populations studied are very close

genetically, are rather homogeneous, and nearly all variation detected has

intrapopulational origin. There is not significant genetic differentiation between them,

despite the geographic barrier posed by Negro River.

Financial support: CNPq and CAPES

P-13

THE PUPAE OF FIVE LUTZOMYIA SPP. (DIPTERA: PSYCHODIDAE) FROM

THE PERUVIAN ANDES.

PEREZ1, J. ENRIQUE, DIANA RADO

2 & ELENA OGUSUKU

3

1 Instituto de Medicina Tropical Alexander von Humboldt Universidad Peruana

Cayetano Heredia. kike@upch.edu.pe. 2 Universidad Nacional San Antonio Abad,

Cuzco. 3 Direccion General de Salud Ambiental, Ministerio de Salud.

The pupae of L. noguchii, L. oligodonta, L. peruensis, L. tejadai and L. verrucarum

from different localities in the Peruvian Andes were studied by means of scanning

electron microscopy.

Adults of sandflies were collected in Chaute, Lima (L. noguchii, L. peruensis),

Cocachacra, Lima (L. oligodonta), Cascay, Huanuco (L. tejadai) and Surco, Lima (L.

verrucarum). Sandflies were fed by exposing of anesthetized mice, engorged females

were individualized in oviposition vials, after oviposition, the females were identified.

The F1 generation was obtained by rearing these species in our laboratory colony. The

pupae were separated, killed in water at 70 ºC, and fixed and freeze dehydrated in 5

passages in absolute ethanol. The material was processed for scanning electron

microscopy in the Department of Pathology, University of Texas Medical Branch,

Galveston, Texas, USA.

The median longitudinal crest was studied in detail for species differentiation. Two

types of median longitudinal crest were observed in the pupae studied, these are the “Y”

shape (cf. L. longipalpis) in L. verrucarum, and a new morphology which resembles a

“V” in all the other species. This “V” was observed in L. noguchii, L. peruensis, and L.

tejadai, which belong to the Subgenus Helcocyrtomyia, and in L. oligodonta

(ungrouped species). The “V” shape of the Subgenus Helcocyrtomyia and L. oligodonta

pupae can be used as a discrimination tool from L. (Lutzomyia) longipalpis and L.

verrucarum.

P-14

MOLECULAR AND BEHAVIORAL DIFFERENTIATION AMONG

BRAZILIAN POPULATIONS OF LUTZOMYIA LONGIPALPIS (DIPTERA:

PSYCHODIDAE: PHLEBOTOMINAE).

ALEJANDRA S. ARAKI1, FELIPE M. VIGODER

1, LUIZ G. S. R. BAUZER

1,

GABRIEL E. M. FERREIRA1, NATALY A. SOUZA

2, IZENEIDE B. ARAÚJO

3,

JOHN G. C. HAMILTON4, REGINALDO P. BRAZIL

5, ALEXANDRE A.

PEIXOTO1

1Laboratório de Biologia Molecular de Insetos, IOC, Fundação Oswaldo Cruz, Rio de

Janeiro, Brazil. 2Laboratório de Transmissores de Leishmanioses, IOC, Fundação

Oswaldo Cruz, Rio de Janeiro, Brazil. 3Curso de Ciências Biológicas e Agrárias,

Universidade Estadual do Piauí, Parnaíba, Piauí, Brazil. 4Centre for Applied

Entomology and Parasitology, Institute of Science & Technology in Medicine, Keele

University, United Kingdom. 5Laboratório de Bioquímica e Fisiologia de Insetos, IOC,

Fundação Oswaldo Cruz, Rio de Janeiro, Brazil.

Lutzomyia longipalpis, the main vector of American visceral leishmaniasis, is a

species complex but until recently the existence of sibling species among Brazilian

populations was considered a controversial issue. In addition, there is still no consensus

regarding the number of species occurring in this complex. We combined molecular and

behavioral approaches to study the differentiation between Brazilian populations of L.

longipalpis. Using period, a gene that controls circadian rhythms and male courtship

songs in Drosophila, we analyzed the molecular polymorphism in a number of L.

longipalpis samples from different regions of Brazil and compared the results with our

previously published data using the same marker. We also studied the male copulation

songs and pheromones from some of these populations. The results obtained so far

suggest the existence of two main groups of populations in Brazil, one group

representing a single species with males producing Burst-type copulation songs and

cembrene-1 pheromones; and a second group that is more heterogeneous and probably

represents a number of incipient species producing different combinations of Pulse-type

songs and pheromones.

Financial Support : This work was supported by grants from the Howard Hughes

Medical Institute, CNPq, Capes, FAPERJ and FIOCRUZ.

II: Biology of Sandflies

P-15

STUDIES ON THE FEEDING HABITS OF LUTZOMYIA (LUTZOMYIA)

LONGIPALPIS (DIPTERA: PSYCHODIDAE: PHLEBOTOMINAE), VECTOR

OF VISCERAL LEISHMANIASIS IN BRAZIL

MARGARETE M S AFONSO¹, ROSEMERE DUARTE2, JOSÉ C MIRANDA

3,

LINDEMBERGH C SOUSA4 & ELIZABETH F RANGEL¹

¹Lab. Transmissores de Leishmanioses, IOC/FIOCRUZ; 2Laboratório de Pesquisa e

Serviços em Saúde Pública, ENSP/FIOCRUZ; 3Laboratório de Imunoparasitologia,

CPqGM/FIOCRUZ; 4Laboratório de Entomologia Médica Dr. Thomaz Correa Aragão,

SESAU-CE, Brasil

Lutzomyia (Lutzomyia) longipalpis, the most important vector of American Visceral

Leishmaniasis (AVL) in Brazil, is adapted to different environments being found in

several habitats including domiciles in urban areas. Data on its feeding behavior are still

not enough and new studies may provide better understanding of the urbanization of the

disease. The aim of this study is to identify the feeding blood sources of L. (L.)

longipalpis from Jequié (BA), Teresina (PI), Sobral (CE) and Massapê (CE, Brazilian

areas of AVL transmission. The ingested blood was identified using ELISA test, with

antisera specific for bird, dog, opossum, human and rodent. From 609 tested females,

40.72% were positive. The human antiserum was positive in females populations from

Jequié and Massapê. All populations were reactive for birds and dogs anti-sera; the high

positive percentage for both suggests the important role of these domestic animals in the

adaptation process of the vector to the domicile environment. The reactivity for more

than one blood source confirms the eclecticism feeder pattern of L. (L.) longipalpis,

which contributes for its adaptation to different habitats. For the first time were

identified females of L. (L.) longipalpis fed on opossums, in populations from Jequié,

Sobral and Massapê. Since, Didelphis albiventris have been found infected by

Leishmania (L.) infantum chagasi, the association between L. (L.) longipalpis and

opossums suggests a better evaluation of this sinanthropic mammal’s role on the AVL

transmission cycle.

Financial Support: IOC, ENSP/FIOCRUZ, DECIT-CNPq/2006, CNPq

P-16

THE USE OF CUTICULAR HIDROCARBONS TO DISTINGUISH SIBLING

SAND FLY SPECIES: LUTZOMYIA (NYSSOMYIA) WHITMANI S.L. (DIPTERA:

PSYCHODIDAE: PHLEBOTOMINAE) AS MODEL

ANDREY JOSÉ DE ANDRADE1, ELIZABETH F.RANGEL

2, ÁLVARO

EDUARDO EIRAS1

1Depart. de Parasitologia, Universidade Federal de Minas Gerais, Belo Horizonte,

MG,bioandrey@gmail.com/alvaro@icb.ufmg.br 2Lab.Transmissores de Leishmanioses, Instituto Oswaldo Cruz/FIOCRUZ, Rio de

Janeiro,RJ, Brasil - elizabethrangel@fiocruz.br

Currently four lineages of phlebotomine sand fly Lutzomyia (Nyssomyia) whitmani s.l.

have been suggested in Brazil. These possible sibling species have a large geographical

distribution and play an important role as vector of Leishmania (Viannia) shawi and L.

(V.) braziliensis, in the Amazonian Region and others Brazilian regions, respectively.

Vector insects present wax cuticular hydrocarbons, and there are analyses have showing

that these compounds are apparently species-specific and could act as chemotaxonomic

markers. The aim of the study is to provide evidences about hydrocarbons of sand flies

and to propose the use of hydrocarbon’s characterization, using the “complex” L.

whitmani s.l. as model. Sand flies were caught in the five geographical regions from

Brazil: North (Pará State); Northeast (Ceará and Bahia States), Southeast (Minas Gerais

State), South (Paraná State), and Center-West (Mato Grosso State). Fifty specimens of

each population were separated by sex and were used in the hydrocarbons identification.

The hydrocarbons were extracted with 1 ml of hexane solvent for 20 min., for 2 and 24

hrs. The hexane layer was filtered and dried on a N2 flow. The extracts were diluted in

20ml hexane and aliquots of 2 ml of each sample were injected for Gas

Chromatography coupled Mass Spectrometry (GC-MS). The standardization of the

hydrocarbons extraction technique is under development. The first results of this step

indicated that 20 min. and 50 specimens are the best to the preliminary GC analysis.

However, different amount of males and females will be used for better improving the

GC analysis, before to evaluate each population. There are only six articles in the

literature using hydrocarbons identification and to distinguish sibling species of sand

flies. Probably these chemical analysis would be an important tool to study the

“complex” L. whitmani s.l.

Financial supporte: CNPq

P-17

IDENTIFICATION OF SEX PHEROMONES OF LUTZOMYIA LONGIPALPIS

(LUTZ & NEIVA, 1912) POPULATION FROM ASUNCIÓN, PARAGUAY.

BRAZIL RP

1, HAMILTON JGC

2, CABALLERO NNG

1, CANESE A

3.

1- Lab. de Bioquímica e Fisiologia de Insetos - Instituto Oswaldo Cruz, Brazil

e-mail: rpbrazil@ioc.fiocruz.br 2- Center for Applied Entomology and Parasitology, Keele University, England

3- Facultad de Ciencias Médicas, Universidad Nac. de Asunción, Paraguay

The sand fly Lutzomyia longipalpis (Lutz & Neiva) is the main vector of Leishmania

(L.) infantum (Nicolle), the causative agent of the American Visceral Leishmaniasis

(AVL) in the New World. Male of L. longipalpis have secretory glands which produce

sex pheromone in either abdominal tergites 3 or 3 and 4. These glands give rise to one-

spot (1S) or two-spot (2S) morphology. It has been suggested that these chemotype

populations represent true sibling species. In Latin America, besides Lu.

Pseudolongipalpis arrivillaga and Feliciangeli from Venezuela, populations of

Lutzomyia longipalpis can be identified by their male-produced sex pheromones: (S)-9-

methylgermacrene-B, 3-methyl--himachalene, and 2 cembrenes, 1 and 2.

In this study we present the results of an analysis, by coupled gas chromatography -

mass spectrometry, of the pheromones of males L. longipalpis captured in an endemic

area of AVL in Asunción, Paraguay. Our results showed that L. longipalpis from this

site produce (S)-9-methylgermacrene-B which has also been found in L. longipalpis

found different areas of the Mato Grosso do Sul along the border with Paraguay.

Financial Support: Faperj, Wellcome Trust and Fiocruz

P-18

BLOOD MEAL IDENTIFICATION IN LUTZOMYIA LONGIPALPIS (LUTZ &

NEIVA, 1912) CAPTURED IN THE MUNICIPALITY OF JANAÚBA, STATE

OF MINAS GERAIS, BRAZIL.

MICHALSKY E.M.1,3

, FRANÇA-SILVA J.C.2, LOROSA E.S.

4, LOUREIRO

A.M.F.5, BARATA R.A.

1, DIAS E.S.

1

1Centro de Pesquisas René Rachou/FIOCRUZ-MG,

2Universidade Federal de Minas

Gerais-MG, 3Universidade Federal do Triângulo Mineiro-MG,

4Instituto Oswaldo

Cruz/FIOCRUZ-RJ, 5Secretaria Municipal de Saúde de Janaúba; Brazil.

Introduction: Lutzomyia longipalpis, the main vector of visceral leishmaniasis in

Brazil, is a species well adapted to anthropic environments. Due to the hematophagic

behavior of the females, analysis of their intestinal contents has been usually employed

to determine on which animal species they are feeding in nature. This data might

indicate a possible participation of these animals in the maintenance of the enzootic

cycle of VL and their eventual role as reservoirs, which may help in understanding the

epidemiology of the disease. Objective: To identify the feeding habits of L. longipalpis

females in Janaúba, an area of transmission of VL in Brazil. Methodology:

Entomological captures were performed in 15 districts of the municipality under study

using HP light traps installed inside and outside households. Engorged phebotomine

females captured were dissected for species identification. The individual intestinal

contents of those belonging to L. longipalpis species were macerated in saline and

centrifuged. The supernatants were submitted to precipitin test using antisera from

different animals. Results: Intestinal contents of 73 females of L. longipalpis were

analyzed and single reactivities were obtained for the following antisera: human

(11.0%), equine (13.7%), opossum (5.5%), bovine (4.1%), porcine (4.1%), avian

(21.9%) and rodent (12.3%). Twenty-three percent (24.7%) of the samples displayed

mixed reactivity (feeding in more than one animal) and 2.7% were non-reactive to the

antisera employed. Conclusions: Our results show that females of L. longipalpis are

able to use a variety of animals as their blood source, probably adjusting their feeding

habits to available hosts. This adaptability favors the adaptation of this vector species to

anthropic environments thus contributing to the urban transmission of VL.

FINANCIAL SUPPORT: CNPq, FAPEMIG, FIOCRUZ, UFTM.

P-19

ANALYSIS OF THE DEVELOPMENTAL CYCLE DIFFERENCES AMONG

BRAZILIAN SIBLING SPECIES OF THE LUTZOMYIA LONGIPALPIS

COMPLEX.

NATALY A. SOUZA, CLAUDIA A. ANDRADE-COELHO, VANDERLEI C.

SILVA, RICHARD D. WARD3 & ALEXANDRE A. PEIXOTO2

1

Lab. Transmissores de Leishmanioses & 2

Lab. Biologia Molecular de Insetos,

Instituto Oswaldo Cruz/ Fiocruz, Rio de Janeiro, Brazil; 3 Institute of Science and

Technology in Medicine, School of Life Sciences, Keele University, Staffordshire, U.K.

The establishment of laboratory colonies of phlebotomine sandflies is vital to undertake

various studies including aspects of behaviour which are still poorly understood. In the

present report the laboratory developmental cycles of five Brazilian populations of the

Lutzomyia longipalpis species complex were compared. Three of the populations were

derived from insects collected in 3 allopatric sites at Natal, Rio Grande do Norte,

Jacobina, Bahia and Lapinha, Minas Gerais. The other two originated from a sympatric

site Sobral, Ceará. Each life stage was recorded including egg production and longevity,

larval and pupal development time and the completion rates of adult emergence. The

results clearly showed that all 3 populations in the study whose males produce C16 and

use pulse-type copulation songs (Jacobina, Lapinha, and Sobral 1S) are more easily

adapted to the colonisation conditions used in our laboratory, producing larger egg

batches, with a lower oviposition mortality and having an overall faster developmental

cycle. This contrasts with populations producing C20 male pheromones and using burst-

type copulation songs (Natal and Sobral 2S) that produce smaller egg batches, have

higher oviposition mortality and a slower developmental rate under identical laboratory

conditions. These phenological differences are a further indication of the differentiation

of the siblings within the L. longipalpis complex in Brazil.

P-20

LIFE-CYCLE AND LARVAL MORPHOLOGY OF LUTZOMYIA MARUAGA

ALVES, FREITAS & BARRETT, 2008

RONILDO B. ALENCAR, VERACILDA R. ALVES, TOBY V. BARRETT

Instituto Nacional de Pesquisas da Amazônia, AM, Brazil

Observations on the life-cycle of a troglobiont and apparently extreme endemic sand fly

from one of the rare arenitic caves of the Central Amazon are presented. All 400 adults

of L. maruaga captured in light traps were females. In addition, 520 larvae and pupae

were extracted by flotation from guano from the cave. All of the adults that emerged

were females, which refused to feed on hamster or human blood, as did the individuals

captured in the caves as adults. Adults laid eggs or died with mature eggs retained in the

abdomen. Both laid and retained eggs hatched, to yield another all-female generation of

adults. The development time from laid egg to eclosion was up to 20 days, and from

first-stage larva to adult, another 90. The external pair of caudal bristles in second to

fourth instar larvae of L. maruaga are three to four times shorter than the internal.

Discussion: Parthenogenesis in Phlebotominae has previously been reported only for L.

mamedei. One of the advantages of sexual reproduction is that it promotes genetic

variability, useful for species whose members face natural selection in an inconstant

environment. In the case of L. maruaga, her world may be so predictable as to make

males an unnecessary drain on resources, although any perturbation could lead rapidly

to extinction. L. maruaga is autogenous, as females do not require a blood meal to

procreate. This could be related to the long development time of the larvae in the

nutrient-rich guano, and the evident presence of a bolus in the midgut of teneral adults.

This makes sense, as obtaining a bloodmeal from an insectivorous bat is clearly

hazardous. On the other hand, the mouthparts of L. maruaga appear to be functional.

Perhaps, in nature, the species does feed on blood, after the first oviposition. Most

known larvae of the genera Phlebotomus, Sergentomyia and Lutzomyia, possess a single

pair of cauldal bristles in the first larval instar, and two pairs of approximately the same

length in the subsequent instars. Exceptions include P. tobii, in which the external

bristles are vestigial. L. maruaga shares with P. mirabilis, a cavernicolous African

species, a pronounced reduction of the length of the external caudal bristles. Whether or

not this coincidence can be attributed to a common cretaceous ancestry, or to parallel

evolution, the true function of the caudal bristles remains uncertain.

P-21

HISTOLOGY OF THE ALIMENTARY CANAL OF LUTZOMYIA

(NYSSOMYIA) INTERMEDIA (LUTZ & NEIVA, 1912): (DIPTERA:

PSYCHODIDAE: PHLEBOTOMINAE) LARVAE: VECTOR OF CUTANEOUS

LEISHMANIASIS IN BRAZIL

1,2

WAGNER A. COSTA, 2SIMONE M. DA COSTA,

2ELIZABETH F. RANGEL,

2

JACENIR MALLET, 3JOSÉ EDUARDO SERRÃO

1 Depart. Biologia Animal, Universidade Federal De Viçosa-Mg,

2Lab. Transmissores de

Leishmanioses, IOC/ Fiocruz, RJ; 3Departamento de Biologia Geral, Universidade

Federal de Viçosa-Mg, Brasil.

The sand fly species, vectors of leishmaniases, are identified using gross morphology of

the adulthood, which made the identification of immature stages hard due to scarcity of

data from larvae. This work describes the anatomy and histology of alimentary canal of

Lutzomyia (Nyssomyia) intermedia larvae. In the second instar larvae the foregut is

short and the midgut is an undifferentiated tube, sometimes having one constriction on

the median portion. In the former portion of the midgut, the cells are flattened, followed

by cubic cells in the median portion. In the posterior portion of midgut the cells are

acidophil, with the globular cells apparently containing secretions in apical portion. In

the third instar larvae the pharynx and salivary glands are placed in head being

surrounded by thin layer of muscle cells. The midgut also presents cubic and globular

cells. In the fourth instar larvae it is evident a dilated pharynx, with flattened cells lined

of thick cuticle. The salivary glands are well developed located after the pharynx. The

proventriculus is well developed, but it is narrower near to the cardiac valve. The

midgut is long reaching approximately the abdominal segment IV, where cells are

flattened and some are globular. The hindgut is cylindrical and its wall has high cells.

These results will be compared with those obtained for Lutzomyia (N) neivai in order to

contribute to the identification of sand fly larvae.

III A: Ecology, Distribution, and Population Dynamics of Sandflies

P-22

UPDATED LUTZOMYIA SPECIES IN PERU (DIPTERA-PSYCHODIDAE).

ROBERTO FERNÁNDEZ1, NELSON SOLORZANO

2, ETTY LOPEZ

3 & KIRK

MUNDAL1

1 Naval Medical Research Center Detachment, Lima-Peru. (NMRCD). 2 Ministerio de Salud. Dirección Regional de Salud de Ancash. Hospital de Caraz.

3 Ministerio de Salud. Dirección Regional de Salud de San Martín.

The study of Phlebotomine sand flies in Peru, began in 1913 with the

entomologist Charles Townsend. He worked in the Rimac Valley (Lima), and his main

contribution was the description of Lutzomyia verrucarum, which is the vector of

Carrion’s disease. Since its description, taxonomic studies have made slow progress. In

the 1960s, investigators launched studies to expand the knowledge of the phlebotomine

fauna within Peru. The majority of the work was done in the eastern regions with new

records of species, new descriptions and their distributions being recorded. Since 1986,

sand fly collections have been accomplished in the different regions of Peru, with

collaborations among universities, Regional Health Ministries, and others researchers.

The specimens were identified to species using internal and external morphological

characters of adults, based in Young & Duncan (1994), with many new species being

described. This information along with an extensive literature review was done to

update the species list of Lutzomyia in Peru. We reported 162 species belonging to 13

subgenera, 8 groups of species and 5 species as non grouped. We also reported new

country records of: Lutzomyia andersoni, L. witoto, L. saltuosa, L. pilosa, L. cellulana,

L. nematoducta, L. marienkellei, L. trapidoi, L. velascoi, L. brachipyga and L. tortura,

with 6 new species that still require description.

P-23

EPIDEMIOLOGICAL STUDY OF AMERICAN CUTANEOUS

LEISHMANIASIS (ACL), AT NORTHERN PARANÁ STATE, BRAZIL, IN

AREAS AROUND THE HYDROELECTRIC PLANTS ON THE

PARANAPANEMA RIVER, 2004 -2006.

FORDELLONE-CRUZ MF*, GALATI EAB**, CRUZ CFR*, PAULA.IRP*

UENP/FALM-PR-BR **FSP/USP-SP-BR

INTRODUCTION: There is a great number of areas affected by human activity, such

as farming and the installation of hydroelectric plants in the state of Paraná. However

the arthropods fauna that acts like vectors within infectious illnesses is rarely known.

Among those are the sandflies (Diptera, Psychodidae), vectors of the ACL.

OBJECTIVE: to study the risk factors associated to the presence of sandflies, in order

to evaluate the transmission of ACL. METHODS: The captures of sandflies were

undertaken with automatic light traps, between 18:00-7:00h, at 25 different spots and an

epidemiological questionnaire was applied to 316 people who live in the surrounding

areas (197.301 km2; 402 m.a.s.l.) situated in northern of Parana State (2302’00’’ S and

50 22’00’’ W). RESULTS: A total of 3187 sandflies were captured belonging to 15

species. The standardized abundance index being: Ny. neivai (0.87), Pi. pessoai (0.79),

Ny. whitmani (0.50), Br.brumpti (0.41), Mg. migonei (0.35), Pi. fischeri (0.20), Br. sp

(0.14), Br. cunhai (0.10), Br. nitzulescui (0.07), Ex. firmatoi (0.05), Ev. bacula (0.03),

Si. sordellii (0.02), Mi. ferreirana (0.01), Pa. aragaoi (0.01) and Pa. shannoni (0.01).

Ny. neivai (34.4%) predominated, followed by Pi. pessoai (32.6%), Mg. migonei

(11,6%), Ny. whitmani (8,8%) and Pi. fischeri (2,7%). From the people who answered

the questionnaire, 90% knew nothing about the transmission of the disease.

CONCLUSIONS: Within all the sites, there were found vector species. There are some

forest areas left, as well as domestic and wild animals, nocturnal activities and fishing.

The lack of information about the leishmaniasis stirs up the need of educational

activities in order to prevent the illness. Some measures could be recommended, such as

the use of repellents, as well as to avoid the schedules of vector’s bigger activity. The

collect of vectors in the surrounding areas indicated that they are getting adapted to the

conditions of life in urban environment, which could represent an increase on the

transmission of the illness in these locals.

P-24

EPIDEMIOLOGICAL STUDY OF RISK FACTORS FOR THE

TRANSMISSION OF AMERICAN CUTANEOUS LEISHMANIASIS (ACL), IN

THE ECOLOGICAL PARK “MATA SÃO FRANCISCO”, STATE OF PARANÁ,

BRAZIL, 2005 -2007.

CRUZ, CFR*,FORDELLONE-CRUZ, MF*,GALATI, EAB**

*UENP/FALM-PR-BR **FSP/USP-SP-BR

INTRODUCTION: There is a great number of parks in the State of Paraná, some of

them with little information on sandflies (Diptera, Psychodidae), vectors of the ACL.

OBJECTIVE: To investigate the risk factors associated to the presence of sandflies, in

order to evaluate the transmission of ACL. METHODS: Captures of sandflies were

undertaken with automatic light traps, between 18:00-7:00 h. Epidemiological

questionnaires were applied to people who either visit or live in the park and

surrounding areas. The ecological park “Mata São Francisco” is situated between the

cities of Cornélio Procópio and Santa Mariana, within an area of 832.57 hectares,

covered by part of the Atlantic forest. RESULTS: A total of 501 sandflies were

captured. The standardized abundance index being: Pintomyia pessoai (37.0%),

Migonemyia migonei (25.2%), Pintomyia fischeri (2.8%), Nyssomyia whitmani (4.8%)

and N. neivai (1.2%). The other species: Brumptomyia brumpti (10.7%), Br. sp (5.0%),

Br. cunhai (1.99%), Br.nitzulescui (0.4%), Evandromyia bacula (0.2%), Ev. correalimai

(2.0%) Ev. cortelezzii (0.2%), Expapilata firmatoi (2.6%), Lutzomyia amarali (0.2%),

Micropygomyia ferreirana (0.2%), Pintomyia monticola (0.2%), Psathyromyia aragaoi

(2.0%), Psathyromyia lanei (1.0%), Psathyromyia shannoni (0.6%), Psathyromyia sp

(0.4%) and Sciopemyia sordellii (0.6%). The Females (59.7%) predominated over the

males (40.3% ). From the 102 people who answered the questionnaires, 85% knew

nothing about the vectors of leishmaniasis. CONCLUSIONS: The high predominance

of vector species (71.0%) and the number of females represent a risk factor for the

transmission of the ACL to people who either visit the park or live there. The lack of

information about the leishmaniasis transmission makes it necessary to develop

activities purposed tourism and management organizations, as well as the use of

repellents and long sleeves clothing could be recommended to the visitors, researchers

and staff members of the park.

Acknowledgements: IAP-Ambiental Institute of the State of Paraná. Financial Support: FUNDACIÓN MAPFRE

P-25

EPIDEMIOLOGICAL STUDY OF RISK FACTORS FOR THE

TRANSMISSION OF AMERICAN CUTANEOUS LEISHMANIASIS (ACL), AT

THE ECOLOGICAL PARK OF VILA VELHA, CITY OF PONTA GROSSA,

STATE OF PARANÁ, BRAZIL, 2005 -2007.

FORDELLONE-CRUZ, MF*,GALATI, EAB**,CRUZ, CFR* PAULAIR*/

*UENP/FALM-PR **FSP/USP-SP

INTRODUCTION: The Vila Velha Park in the Paraná State, Brazil, is a great

attraction for tourists. However, the knowledge on sandflies (Diptera, Psychodidae,

Phlebotominae) and vectors of the American cutaneous leishmaniasis (ACL) is at most

inexistent. OBJECTIVE: to study the risk factors associated to the presence of

sandflies, in order to evaluate the transmission of American cutaneous leishmaniasis

(ACL). METHODS: The captures of sandflies were undertaken with automatic light

traps, settled at five different spots, between 18:00-7:00h, from January 2006 to January

2007. An epidemiological questionnaire was applied to the visitors of the park and to

the inhabitants from the surrounding areas. The Park area, with 3,122 hectares, is

formed by sandstones covered with open vegetation and also mixed umbrophylous

forest. RESULTS: A total of 125 phlebotomines belonging to 14 species, and 52

bruchomyiines (Bruchomyia spp., 2 species) were collected. Among the phlebotomines,

Brumptomyia ortizi predominated (27.2%) followed by Lutzomyia gaminarai (22.4%).

The other species captured were: Br. figueiredoi, Br. guimaraesi, Br. nitzulescui, and

Br. troglodytes, Evandromyia correalimai, Ev. edwardsi, Ev. lenti, Expapillata firmatoi,

Migonemyia migonei, Pintomyia fischeri, Pi. monticola and Psathyromyia lanei. From

the 201 people who answered the questionnaire, 90% knew nothing about the vectors of

leishmaniases. CONCLUSIONS: Only two potential vectors of cutaneous

leishmaniasis were found, Mg. migonei (3.95%) and Pi. fischeri (3.39%). Due to the

very low frequencies the transmission rate of this disease within the area is low. The

find of Lu. gaminarai, species belonging to the same monophyletic group of Lu.

longipalpis, implies the need of more bio-ecology researches about its potentiality as a

visceral leishmaniasis vector. Some preventive measures can be established, such as the

use of repellents for visitors, staff members, researchers and trainees. It could also be

recommended to be developed a health educational work, in order to explain about the

illness. Acknowledgements: IAP-Ambiental Institute of the State of Paraná. FINANCIAL SUPPORT: FUNDACIÓN MAPFRE

P-26

PRELIMINARY RESULTS ON POPULATION DISPERSAL STUDIES OF

SANDFLIES (DIPTERA: PSYCHODIDAE: PHLEBOTOMINAE) SUPPORTED

BY GEOTECHNOLOGIES IN AN URBAN ENDEMIC AREA OF VISCERAL

LEISHMANIASIS OF MATO GROSSO DO SUL, BRAZIL

EVERTON FALCÃO DE OLIVEIRA1, ANTONIO CONCEIÇÃO PARANHOS

FILHO2, ELAINE ARAÚJO E SILVA

3, MARIA ELIZABETH CAVALHEIROS

DORVAL1, REGINALDO PEÇANHA BRAZIL

4, VALTER ALEXANDRE

TIVIROLI2, ALESSANDRA GUTIERREZ DE OLIVEIRA

1.

1. Universidade Federal de Mato Grosso do Sul - Laboratório de

Parasitologia/DPA/CCBS, Campo Grande, MS, Brazil. 2. Universidade Federal de

Mato Grosso do Sul - Laboratório de Geoprocessamento para Aplicações

Ambientais/DHT/CCET, Campo Grande, MS, Brazil 3. Centro de Controle de

Zoonoses, Secretaria de Saúde do Município de Campo Grande, MS, Brazil. 4. Instituto

Oswaldo Cruz- LBII, Rio de Janeiro, RJ, Brazil.

Dispersal of sandflies has been studied in the city of Campo Grande, MS, Brazil. A

preliminary experiment was performed in January 2008 in a district of the south of the

city, in which insects were captured, marked with a fluorescent powder released and

recaptured. The initial collection of insects was performed inside a chicken coop. The

insects were collected with an electrical aspirator between 06:00 and 11:00 PM. The

captured specimens were then marked and released on the same night. The attempts of

recapture were made with CDC light traps installed in sites around the house were

insects were released and in 19 geocoded points in a range of 300m around the released

point, chosen with the aid of a geoprocessing program. Recaptures were performed over

seven consecutive nights following the release. A number of 542 insects were caught,

marked and released from which 17,89% (97 insects, 75 males and 22 females) were

recaptured. The analysis of the points showed the spatial distribution of the recaptured.

The maximum distance of dispersion was near 223m reached by a female in the second

day after release. From the marked insects, 75,3% were recaptured at the release site,

9,3% at 156m, 10,2% at 169 and 5,2% to a distance of 223m from the release site.

P-27

ECOLOGICAL ASPECTS OF SANDFLIES (DIPTERA: PSYCHODIDAE:

PHLEBOTOMINAE) IN CAMPO GRANDE, VISCERAL LEISHMANIASIS

ENDEMIC AREA OF MATO GROSSO DO SUL, BRAZIL

ALESSANDRA GUTIERREZ DE OLIVEIRA1, REGINALDO PEÇANHA

BRAZIL2, ITALO ALEXANDER CABELLO ESPINDOLA

3, MARIA

ELIZABETH CAVALHEIROS DORVAL1, CARLOS EURICO FERNANDES

1,

EUNICE APARECIDA BIANCHI GALATI4.

1. Universidade Federal de Mato Grosso do Sul, Laboratório de

Parasitologia/DPA/CCBS, Campo Grande, MS, Brazil, E-mail: agoliveira@nin.ufms.br.

2. Instituto Oswaldo Cruz- LBII, Rio de Janeiro, RJ, Brazil. 3. Acadêmico do curso de

Farmácia/Universidade Católica Dom Bosco-UCDB, Campo Grande, MS, Brazil. 4.

Universidade de São Paulo, FSP, Departamento de Epidemiologia, São Paulo, Brazil.

Aspects of ecology of sandflies in Campo Grande, MS were studied. With the aid of

CDC light traps, insects were captured weekly in seven different sites including forest

remains and houses from December 2003 to November 2005. Twenty five species were

identified in the areas studied. From a total of 11,024 specimens captured (7,805 males

and 3,219 females), 9,963 (90,38%) were identified as Lutzomyia longipalpis, the

proven vector of visceral leishmaniasis; the remaining specimens were distributed

among the other 24 species. The most frequent species in all sampled areas was Lu.

longipalpis, and this species was the first on the ranking of the padronized abundance

index; it was also the predominant species caught in areas surrounding human dwellings

(90,96%) in contrast to a much lower percentage inside the houses (9,04%). In animal

shelters it was found predominantly in chicken coops and prevailed at land level into

and at the border of the forest remains near the housing areas. This finding is of great

importance as this site may serve as scape areas for the species from the inside house

spraying. Into the areas of remains of forest the following species vector of cutaneous

leishmaniasis were also found: Nyssomyia whitmani, Ny. antunesi and Bichromomyia

flaviscutellata.

Financial support: FUNDECT 498/02, DECIT/FUNDECT, process 41/100208/2003

and FIOCRUZ.

P28

STUDIES ON SAND FLY FAUNA (DIPTERA:PSYCHODIDADE) FROM

TRANSMISSION AREAS OF AMERICAN CUTANEOUS LEISHMANIASIS IN

STATE OF ACRE, BRAZIL.

ALFREDO CR AZEVEDO, SIMONE M COSTA, MARIAC G PINTO*, JANIS L

SOUZA*,HENRIQUE C CRUZ*, JOAQUIM VIDAL*, ELIZABETH F RANGEL

Lab. Transmissores Leishmanioses, Instituto Oswaldo Cruz/FIOCRUZ, Rio de Janeiro,

RJ, Brazil * Secretaria de Saúde do Estado do Acre, Rio Branco, Brasil

Amazonia has an American Cutaneous Leishmaniasis (ACL) epidemiological circuit

with rural and occupational characteristics, associated with the forest destruction, in the

State of Acre. The aim of the study is to know the sand fly fauna and vectors of ACL in

Acre with particular interest in Lutzomyia (Nyssomyia) umbratilis. Sand fly collections

were done in the Municipalities of Bujari (Area I), Xapuri (Area II) and Rio Branco

(Area III, Area IV and Area V), from 2002 through 2006. Two of the localities (Bujari –

Area I and Rio Branco – Area III) were considered strategic areas for regular

collections, considering that these municipalities have been registering high incidence

of ACL human cases. The phlebotomines were collected in anthropic impact forested

areas using CDC light traps. Captures in soil level (50cm) were conducted in Areas I

and III, as well as in the tree canopy (15 – 18m) in Area III. The SISA was used to

calculate data obtained in Areas I and III. During four years 7,302 specimens were

captured in 915 hours. Fifty two species were identified, these Lutzomyia (N.) antunesi,

L. (N.) whitmani, L. (P.) davisi, L. migonei, L. (N.) umbratilis, L. (N.) flaviscutellata, L.

(T.) ubiquitalis, L. (P.) hirsuta hirsuta, L. (P.) paraensis and L. (P.) ayrozai, are vectors

of ACL. L. (N.) umbratilis and L. (N.) whitmani occurred in three municipalities.

Considering systematic collections, L. (N.) umbratilis was the fifth more abundant

species, being more frequent at the Municipal Park of Chico Mendes in Rio Branco; the

vertical stratification registered L. (N.) umbratilis in two levels, predominating in the

canopy of the trees. This study has been suggesting three transmission cycles of ACL in

Acre: Leishmania (V.) braziliensis by Lu. (N.) whitmani, Leishmania (V.) lainsoni by

Lu. (T.) ubiquitalis and, for the first time in the south of the Amazon River transmission

of Leishmania (V.) guyanensis by Lu. (N.) umbratilis.

Supported by: FIOCRUZ, CNPq-DECIT/2006

P-29

SURVEY OF PHLEBOTOMINE SANDFLY FAUNA (DIPTERA:

PSYCHODIDAE) OF AL-ASABA TOWN IN THE NEFUSA MOUNTAIN OF

LIBYA: AN ENDEMIC AREA OF ZOONOTIC CUTANEOUS

LEISHMANIASIS.

EL-ABAIED, *B. ANNAJAR & A. EL-BUNI

Libyan National Centre for Infectious Diseases Prevention & Control

*Correspondence: B. Annajar (bbannajar@yahoo.com)

Zoonotic cutaneous leishmaniasis (ZCL) has been endemic in the Northwestern region

of Libya for over nine decades. Survey of sandfly fauna in the region revealed that 13

species have been recorded with various distribution and abundance patterns.

Phlebotomus papatasi proved to be the main vector of the disease in many areas.

Objective: To identify sandfly species present in the Al-Asaba town and determine

their spatial and seasonal abundance. An epidemiological analysis of the data obtained

from the recorded cases was also carried out.

Methods: Sandflies collected from various sites using sticky traps and CDC miniature

light traps during the period from March – November 2006. Recorded ZCL cases were

collected from the local Primary Health Care Department and analysed using SPSS

statistical package.

Results: Ten species of sandflies were identified, 7 belong to the genus Phlebotomus

and 3 belong to the genus Sergentomyia. P. papatasi was the most abundant species

with peak season recorded in September. The prevalence of the disease was low

however; notable increase of ZCL cases in last 3 years has been indicated.

P-30

STUDY FAUNA OF SAND FLIES IN ENDEMIC AREA FOR AMERICAN

TEGUMENTARY LEISHMANIASIS, MUNICIPALITY OF GOVERNADOR

VALADARES (MINAS GERAIS, BRAZIL).

BASTOS MC1, DIAS ES

1, OLIVEIRA RCA

1, MICHALSKY EM

1, PINHEIRO

AC2, AMBRÓSIO VO

2, LOPES MOG

3, PAZ GF

1, & BARATA RA

1.

1Centro de Pesquisas René Rachou/FIOCRUZ;

2. Secretaria Municipal de Saúde de

Governador Valadares; 3. Biodiversity Salvation

Introduction. In Brazil, the American Cutaneous Leishmaniasis (ACL) have increased

their importance in the context of public health due to the urbanization process,

characterized not only by geographical expansion, invading indenes areas, but also by

the appearance in outbreaks old. The municipality of Governador Valadares was

considered an endemic area for the disease, showing in the last 4 years (2004 to 2007),

221 new cases in accordance with the SINAN. Objectives. To help us understand how

the transmission of ACL may be occurring in the area, this study aimed to know the

wildlife phlebotomine and some aspects of the behaviour of the species for elucidation

of the epidemiology of the disease. Methods. Captures were made with systematic use

of light traps HP during the months of January and February 2008. Sixteen traps were

exposed in 4 districts (Elvamar, Vila Isa, Village da Serra, Vila Parque Ibituruna),

inside the home and outside, for 3 consecutive nights in each month. Results. 2220

specimens were collected, and 1338 females and 882 males, with a share of 32.8% and

67.2% no intra and peridomiciliary, respectively. The sand flies showed the following

distribution: Brumptomyia sp. (0.2%), Lutzomyia cortelezzii (23.3%), L. intermedia

(18.2%), L. ischyracantha (17.1%), L. lenti (9.4%), L. longipalpis (10.0%), L.

quinquefer (18.5%), L. sordellii (0.1%), L. termitophila (1.4%), L. whitmani (1.4%) e

Lutzomyia spp. (0.4%). The neighborhoods Vila Isa and Village da Serra were those

who had the highest occurrence of specimens captured. Conclusions. The results

showed that the fauna phlebotomine in the region is quite diversified with some species

of epidemiological interest. The presence of L. intermedia and L. whitmani strengthen

their roles as a vector of ACL. The meeting of L. longipalpis is worrying, requiring

entomological surveillance for the prevention of kala-azar in the region. The high

percentage of sand flies inside the home points to a high rate of synanthropy and the

possibility of transmission of Leishmania.

Financial support: FIOCRUZ, FAPEMIG, SMS/Governador Valadares.

P-31

PHLEBOTOMINAE (DIPTERA: PSYCHODIDAE) FAUNA IN THE LOW

LAND REGION OF BOLIVIA IN THE BORDER WITH CORUMBÁ, BRAZIL.

BRAZIL RP1, LANÇA PASSOS W

1, TEMELJKOVITCH M

2, TEDESQUI V

1,

ANDRADE FILHO JD3

1- Lab. de Bioquímica e Fisiologia de Insetos - Instituto Oswaldo Cruz, Rio de Janeiro,

Brazil e-mail: rpbrazil@ioc.fiocruz.br 2 -

Secretaria de Agricultura-Prefeitura de Corumbá, MS,Brazil. 3 – Instituto René Rachou, Fiocruz,Belo Horizonte,MG,Brazil

In Bolivia, the incidences of American Cutaneous Leishmaniasis (ACL) and Visceral

Leishmaniasis (VL) have shown to be increasing over last few decades. The low lands

known as the Chaco biogeographical region have extended areas which have borders

with Argentina, Paraguay and Brazil. Both ACL and VL are found in the borderline

areas of the three countries and possible also in Bolivian Chaco. Although several

aspects of AVL and VL are reported from the highland areas, little is known about the

diseases and sand fly composition in the eastern Chaco region of Bolivia. A preliminary

account is shown from captures in peridomestic sites of the town of El Carmen (S 180

49´64´´; W 580 36´95´´) in March 2004 using CDC light traps. Five species of sand flies

were captured: Lutzomyia cruzi, L. forattinii, Evandromyia corumbaensis,

Psathyromyia shannoni and Expapillata cerradincola, with high prevalence of L. cruzi.

Except for P. shannoni all the other species are new records for Bolivia. L. cruzi and L.

forattinii are suspect vectors of Visceral leishmaniasis in Corumbá, Mato Grosso do Sul

just 200 kms of distance with a potential risk of VL transmission in this region of

Bolivia.

Financial Support: Fiocruz and Faperj

P-32

IS LUTZOMYIA CRUZI (MANGABEIRA, 1938) (DIPTERA: PSYCHODIDAE)

PRESENT IN THE STATE OF CEARÁ, BRAZIL?

JOSÉ DILERMANDO ANDRADE FILHO, SOFIA BERNARDES LOURENÇO

BARBOSA, CRISTIANI DE CASTILHO SANGUINETTE, ALDA LIMA

FALCÃO

Laboratório de Leishmanioses - Centro de Pesquisas René Rachou - Fiocruz

The leishmaniases are a group of diseases caused by protozoa of the genus Leishmania

and are transmitted through the bite of female phlebotomines. They can present itself by

two basic clinical forms: American Tegumentary Leishmaniasis (ATL) and Visceral

Leishmaniasis (VL), being the last one lethal, if not treated. The VL presents as the

main vector, in Brazil, the species Lutzomyia longipalpis, meantime the species

Lutzomyia cruzi can also be involved in transmission of this disease. Regarding the

geographic distribution of these species of phlebotomines, both are widely found in

Central West Region of Brazil while in the Northeast Region, L. longipalpis appears in

all states and L. cruzi was recorded only for the State of Ceará [Cipa Group, available in

http://cipa.snv.jussieu.fr/; Aguiar & Medeiros, 2003. Distribuição e Hábitats. In:

Flebotomíneos do Brasil (Rangel & Lainson Eds.), pp.207-257.]. This work aims to

discuss about the discovery of L. cruzi in Ceará using specimens from the Municipality

of Quixadá, deposited in the Collection of Phlebotomines of the Centro de Pesquisas

René Rachou/Fiocruz. These specimens are represented by a female, mounted

individually, identified as L. cruzi and by five males mounted on two slides: one with

three specimens, one L. cruzi and two L. longipalpis; and another with two specimens

determined as L. cruzi. According to one of us (ALF) the information passed to the Cipa

Group to register the specie in Ceará is from such material. After examining these slides

in optical microscope, it was noted the presence of only one species, L. longipalpis,

making void the occurrence of L. cruzi in Ceará, restricting its geographical distribution

to the states of Central West Region. It is interesting to note that the specimens that

were erroneously identified are in perfect condition, being possible to perform, safely,

its identification.

Financial Support: PIBIC/FIOCRUZ/CNpQ, FAPEMIG

P-33

TEMPORAL PATTERNS OF ABUNDANCE OF THE SAND FLY LUTZOMYIA

LONGIFLOCOSA IN A DOMESTIC FOCUS OF AMERICAN CUTANEOUS

LEISHMANIASIS IN CHAPARRAL, TOLIMA, COLOMBIA

DAIRO MARÍN1,2

, CRISTINA FERRO1,3

, LEONARD MUNSTERMANN4,

CLARA OCAMPO1

1 Centro Internacional de Entrenamiento e Investigaciones Médicas, CIDEIM., Cali,

Colombia 2 Laboratorio de Investigaciones en Parasitología Tropica,l LIPT, Ibague, Colombia

3 Investigadora Emérita Instituto Nacional de Salud, INS, Bogota, Colombia.

4 Yale University School of Public Health, New Haven, Connecticut, USA

Introduction: The sand fly Lutzomyia longiflocosa, is considered the principal vector of

cutaneous leishmaniasis (CL) in the Subandean region of Colombia. In 2004, this

species was the most abundant species found in the municipality of Chaparral during the

largest CL outbreak ever recorded in the country.

Objective: to identify temporal abundance patterns of Lutzomyia longiflocosa during

high and low precipitation periods, and determine temporal risk of CL in the

municipality of Chaparral.

Methodology: Sampling was done in the vereda (rural township) of Agua Bonita which

registered high cumulative incidence during the CL outbreak. CDC light traps were set

up monthly in 3 houses for 3 consecutive nights for 11 months beginning in June 2007.

The three houses were selected on the basis of previous observation of high indoors

sand fly abundance. Five traps were set per house: one indoors, 2 peridomestically (10

m away from house) and 2 in neighboring woodland. Preliminary analysis of the

resulting sand fly abundance data was by linkage to local climatic data from the

Instituto de Hidrología, Meteorología y Estudios Ambientales de Colombia (IDEAM).

Results: 59 % of the sandflies (total 10,406) were collected in August and February,

months with lower precipitation. Sand fly collections were much higher in the

peridomestic (42.23 per trap-night ) and woodland environment (36.30) than indoors

(8.87). Lutzomyia. longiflocosa was the most abundant species collected . Cumulative

rainfall over the 15 days before collection, was negatively associated with sand fly

abundance, as was humidity.

Conclusion: In Chaparral, as elsewhere in the Subandean region, the highest sand fly

abundance occurs during the low precipitation period. From a public health perspective,

these data identify the time of highest risk of CL transmission.

POSTER SESSION II

III B: Ecology, Distribution, and Population Dynamics of Sandflies

P-34

NEW EVIDENCE OF LUTZOMYIA LONGIFLOCOSA (DIPTERA:

PSYCHODIDAE) AS VECTOR IN THE UPPER MAGDALENA RIVER

VALLEY COLOMBIA

, LEONARD

1,3DAIRO MARÍN,

,

1O CADENA, HORACI

1,2CRISTINA FERRO

MUNSTERMANN4,CLARA OCAMPO

1

1 Centro Internacional de Entrenamiento e Investigaciones Médicas, CIDEIM., Cali,

Colombia 2 Investigadora Emérita Instituto Nacional de Salud, INS, Bogota, Colombia.

3 Laboratorio de Investigaciones en Parasitología Tropica,l LIPT, Ibague, Colombia

4 Yale University School of Public Health, New Haven, Connecticut, USA

Introduction. In 2004 in Chaparral County, Tolima Province, (elev. 1,000-2,000 m)

the largest recorded outbreak of cutaneous leishmaniasis (CL) in Colombia was

reported. Prevalence in townships(rural sectors) of similar altitude varied from 1% to

95%. Lutzomyia longiflocosa was considered the most probable vector because of its

predominance, high anthropophily and apparent endophagic behavior and Leishmania

guyanensis as most probable agent.

Objective. In order to understand the variation in prevalence within townships of the

municipality of Chaparral of the similar altitude, in two of them we studied the

composition of Lutzomyia and habitat, one with high and one with low prevalence.

Methods. Sampling was undertaken at stations that were defined by a household

located at the center of four radiating transects. Sandflies were collected with CDC light

traps placed inside dwelling and along the transects at 10, 30, 60 and 90 m. intervals

from the household. The type of habitat was also recorded: secondary forest, coffee

plantations, grassland or others.

Results. Twelve sampling stations in the townships of Agua Bonita (AB, with CL

prevalence 74%) and IrcoDos Aguas (ID, with CL prevalence 1.3%) indicated the

presence of many more sandflies in AB (n=1,249) than in ID (n=76). Lutzomyia.

longiflocosa was the most frequent species in AB (78%) and in ID (49%). In AB the

abundance of Lutzomyia was highest. Ten species of sand fly were present in AB and 7

in ID. With regards to associated habitat, AB sites tended to be located adjacent to

forest and shaded coffee plantations, and with little grassland,

Conclusions These differences explain in part the variation in prevalence within the

townships (rural sectors) of the municipality of Chaparral.

P-35

CLIMATE VARIABILITY AND LUTZOMYIA SPINICRASSA ABUNDANCE IN

AN AREA OF CUTANEOUS LEISHMANIASIS TRANSMISSION IN NORTE

DE SANTANDER, COLOMBIA

FREDY GALVIS OVALLOS,1 YANIS RICARDO ESPINOSA,

1 REINALDO

GUTIÉRREZ MARÍN,1 NELSON FERNÁNDEZ,

2 ALFONSO RODRIGUEZ-

MORALES,3 CLAUDIA MAGALY SANDOVAL.

1

1Laboratorio de Entomología Médica, Instituto de Ciencias Biomédicas INBIOM,

Universidad de Pamplona, Colombia; 2Centro de Hidroinformática, Recursos Naturales,

Univ de Pamplona, Colombia; 3Instituto Experimental JWT, Universidad de Los Andes,

Trujillo, Venezuela.

Introduction: Cutaneous leishmaniasis transmission is the product and interplay of

different ecological, biological and epidemiological elements. Recently climate

variability is one ecoepidemiological element influencing that dynamics with a

heterogeneous impact according time and space.

Objectives: For these reasons we have been interested on analyze the impact of climate

variability elements on the abundance of Lutzomyia spinicrassa in an area of cutaneous

leishmaniasis transmission in Norte de Santander, Colombia between February 2006

and February 2007.

Methods: Collection methods include human landing, intra and peridomiciliarly, as

well with Shannon tramps and CDC light tramps. The relation between the sandflies

abundance and climate elements was done taking in consideration a time lag of 5 days,

analyzing relative humidity, average temperature and rainfall. Statistical analysis was

done through the use of main components analysis (MCA) with 95% level of

confidence.

Results: During the study period 5,079 sandflies were collected, L. spinicrassa

represented 95.2% of them. The climatic period corresponded to a dry season of El Niño

(highest Oscillation Niño Index in the last 2006 trimester, 1.2 SST). In general, the

MCA evidenced a significant inverse relation between L. spinicrassa abundance and the

relative humidity (p<0.05), as well also with the rainfall (p<0.05), but not for the

average temperature (p>0.05).

Conclusions: The climatic variability seen in Norte de Santander, in concordance with

the macroclimate variability indicating the shifting to El Niño phase, suggest a suitable

environment for the presence and abundance of L. spinicrassa in this northern

Colombian area, which is in support of previous published data indicating that during El

Niño phases with a dry seasons and environments, sandflies, and eventually

transmission and cutaneous leishmaniasis cases, are prone. These results will provide

important information in the development of tools for forecasting, prevention, early

detection and control of cutaneous leishmaniasis in endemic areas.

P-36

ENTOMOLOGICAL ASPECTS AND CANINE INFECTION IN AREA OF

INTENSE TRANSMISSION OF VISCERAL LEISHMANIASIS IN JANAÚBA,

MINAS GERAIS.

LARA-SILVA, F.O.1, MICHALSKY, E.M.

1, 3, FRANÇA-SILVA, J.C.

2, BARATA,

R. A.1, SOARES, L. F.

1, DIAS, E. S.

1

1 – Centro de Pesquisas René Rachou / Fiocruz

2 – Universidade Federal de Minas Gerais

3 – Universidade Federal do Triângulo Mineiro

Introduction: The transmission of leishmaniasis occurs by the bite of sand flies of the

genus Lutzomyia in the New World. Some mammals act as sources of infection for the

vectors, but the dogs are prosecuted as domestic reservoir of the disease. The Visceral

Leishmaniasis stands out as a public health problem; studies clarify the biology and

behavior of the vector associated with the epidemiological role of the dog, can drive

new discoveries for the control of leishmaniasis. Objectives: Raise the fauna

flebotominica, study their behaviour in relation to endofilia and exofilia and estimate

the prevalence of canine infection in the city. Methodology: Entomological captures

were held monthly, for three consecutive nights using light traps HP, and installed

inside the home, April 2005 to March 2007, in 15 neighborhoods of the municipality.

The canine survey was conducted in the neighborhoods under study in 2006, in order to

estimate the prevalence of infection. The diagnosis used was the reaction of indirect

immunofluorescence in eluate of blood dissected on filter paper. Results: 14591

specimens from sand flies were caught, distributed on 8 species: Lutzomyia longipalpis

(92.39%), L. sallesi (0.63%) and L. intermedia (0.74%), L. lenti (4.14%), L. sordellii

(0.03%), L. termitophila (0.05%), L. whitmani (0.03%), Lutzomyia spp. (1.99%),

Brumptomyia brumpti (0.01%). In relation to the behaviour of the species, 17% were

caught inside the home and 83% in the peridomicile. Of the 6108 dogs examined, 1084

were positive, with average prevalence rate of around 18%. Conclusions: The fauna

flebotominica had to be quite diverse, with the presence of vector species of

leishmaniasis, with L. longipalpis the predominant species. The high rate of prevalence

of canine visceral leishmaniasis, and many sand flies trapped inside the home can

contribute in the urban cycle of transmission in the municipality of Janaúba.

Financial Support: FAPEMIG, UFTM, FIOCRUZ.

P-37

GEOGRAPHICAL DISTRIBUTION OF THE CORTELEZZI COMPLEX

(DIPTERA: PSYCHODIDAE: PHLEBOTOMINAE FROM BRAZIL).

GUSTAVO MAYR DE LIMA CARVALHO1, REGINALDO PEÇANHA

BRAZIL2, ALDA LIMA FALCÃO

1, JOSÉ DILERMANDO ANDRADE FILHO

1.

1- Laboratório de Leishmanioses - Centro de Pesquisas René Rachou - Fiocruz

2- Laboratório de Bioquímica e Fisiologia de Insetos - Instituto Oswaldo Cruz - Fiocruz

Around 500 species of sandflies are known in the Americas and in Brazil more than 300

species are found in the Brazilian territory where about 30 species are proven or

suspicion to transmit leishmaniases. The species Evandromyia cortelezzii, Evandromyia

sallesi and Evandromyia corumbaensis are part of the cortelezzii complex. These

species are morphologically alike and many times they are taxonomically confused,

generating errors in its specific identification. Due to epidemiological importance

recently attributed to this complex we made a detailed study in order to correct the

mistakes committed during the specific identification of these species and update their

geographical distribution based in the recent papers and in these species deposited in the

Collection of the Instituto René Rachou/Fiocruz. It was found that E. corumbaensis has

a more focal distribution, occurring only in Center-West Region, where the others two

species also occurs. E. cortelezzii and E. sallesi is reported in many municipalities of

Minas Gerais State, with the former also reported from the North (Acre and Tocantins

States), Northeast (Bahia, Ceará, and Maranhão States), Southeast (Espírito Santo, Rio

de Janeiro, and São Paulo States) and South Regions (Paraná and Rio Grande do Sul

States) and the later is also found in Rio de Janeiro, Rio Grande do Norte (Northeast

Region), Sergipe (Northeast Region) and Tocantins States. It is expected that this work

shows the necessity of a better morphological study and correct specific identification

before any publications showing a wrong geographical distribution.

Financial Support: IRR/FIOCRUZ, FAPERJ, FAPEMIG

P-38

SAND FLY SURVEY IN THE INFLUENCE AREA OF PEIXE ANGICAL

HYDROELETRIC PLANT, STATE OF TOCANTINS, BRAZIL

VILELA ML, AZEVEDO ACR, COSTA SM, COSTA WA, MOTTA-SILVA D,

GRAJAUSKAS AM, CARVALHO BM, BRAHIM LRN, KOZLOWSKY D &

RANGEL EF.

Lab. Transmissores Leishmanioses, Instituto Oswaldo Cruz/ FIOCRUZ, Av.Brasil,

4365, Rio de Janeiro, RJ, Brasil.

Leishmaniases are emergent diseases with occurrence in all geographic regions of

Brazil. Continuous activities such as deforestation have been modifying the

environments and contributing to the increase of epidemic outbreaks and expansion of

the diseases. The State of Tocantins has been suffering environmental impacts related to

the construction of hydroelectric plants, agriculture activities mostly related to soybean

plantations, and the establishment of new settlement areas which are determining

changes in the landscape. Because of this, leishmaniases have expanded their range and

nowadays an expressive number of municipalities from the State of Tocantins, where

the disease was not present in the past, have now human cases. Some municipalities are

considered highly endemic for both American visceral leishmaniasis (AVL) and

American cutaneous leishmaniasis (ACL), whose have re-emerged in most of the

municipalities. The aim of this study was to identify potential Leishmania vectors and

transmission cycle patterns of ACL in the influence area of Peixe Angical Hydroelectric

Plant., in the South of State. Sand fly captures were conducted with light traps, CDC-

like and Shannon trap, in peridomicile and in the forest, in Paranã, Palmeirópolis, Peixe

and São Salvador do Tocantins municipalities, from February 2004 to May 2008. The

phlebotomine fauna was composed by 54 sand fly species, belonging to the genera

Lutzomyia and Brumptomyia, with a total number of 14,761 collected specimens.

Among the most representative species, Lutzomyia whitmani and L. longipalpis were

found. The occurrence of L. longipalpis is being monitored related to its frequence.

Transmission cycles of ACL are suggested: Leishmania (V.) braziliensis by L. (N.)

whitmani and Leishmania (L.) amazonensis by Lutzomyia (N.) flaviscutellata. Studies

on the search of natural infection by Leishmania spp.in these ACL vectors are in

progress and would give us evidences on the participation of both in the suggested

cycles.

Support: ENERPEIXE, FIOCRUZ, CNPq-DECIT/2006

P-39

SAND FLY (DIPTERA: PSYCHODIDAE) OF AN ENDEMIC LEISHMANIASIS

AREA IN THE ORINOQUÍA - AMAZONÍA OF COLOMBIA

OLGA L. CABRERA1, LAUREANO MOSQUERA

2, ERIKA SANTAMARÍA

1,

CRISTINA FERRO1

1 Grupo de Entomología, Instituto Nacional de Salud, Bogotá, D.C., Colombia.

2 Secretaría de Salud del Guaviare, San José del Guaviare, Colombia.

Introduction. So far, the report of cases of leishmaniasis in the department of Guaviare

did not include entomological studies to identify sand flies vectors

Objective. Identification of species of Lutzomyia in the four townships of Guaviare and

determination of probable ones involved in transmission from their previous record as

vectors.

Materials and methods. Sampling were carried out with CDC light traps suspended

between 1.5 m and 9 m high. In addition sand flies were also caught with Shannon trap

and in resting sites.

Results. Sand flies belonging to 37 different species were collected. 35 of them were

recorded for the first time in Guaviare and 4 for Colombia: L. begonae, L. campbelli, L.

sericea and L. nematoducta. The most abundant species were L. hirsuta hirsuta 24.3 %

(148/610), L. yuilli 15.2 (93/610), L.davisi 10.3 % (63/610), following by L. fartigi, L.

carrerai carrerai, L. antunesi, L. flaviscutellata and L. olmeca bicolor.

Conclusion. Seven species of Lutzomyia were identified as possible sand flies vectors.

P-40

PHLEBOTOMINAE IN NORTHWESTERN ARGENTINA: PRELIMINAR

DISTRIBUTION MAP MODEL

QUINTANA MG 1,3,4

; SALOMON OD 2,3,4

; LIZARRALDE DE GROSSO MS 1,3

1INSUE, National University of Tucumán (UNT),

2: Ministry of Health,

3CONICET,

4REDILA: National Network for the Research of Leishmaniasis in Argentina

gabrieladelaquintana@gmail.com

In the last 20 years, the evidence of an association between vector-borne diseases re-

emergence and environmental changes increased. During the same period the cases of

cutaneous leishmaniasis in Argentina rose associated with changes in vectors density or

distribution, that facilitate their effective contact with humans. The re-emergence was

explained mainly by: 1) deforestation (logging, expansion of the agricultural border), 2)

unplanned urban growth (rural-periurban), 3) climate / environmental factors. In the last

decade, the use of algorithms to model ecological niches of the species was used for

several applications including eco-epidemiology. These models are applied to explore

relationships between the values of biotics (type of vegetation) and abiotic factors

(temperature, precipitation, etc), with data presence, presence-absence and / or

abundance of species. In this study we developed a model to estimate the more uniform

distribution of Phlebotominae in Argentinian Northwestern with the program

MAXENT, with the aim of generating a map of potential distribution of vectors based

on biotic and abiotic conditions, and abundance and distribution of vectors in different

environments of the region, to contribute to design spatially focused strategies for

surveillance and prevention of leishmaniasis. The study area is classified as subtropical

humid forest, where the forest degradation has been taking place since the decade of 80.

The model was developed with 98 entries of Phlebotominae, according to altitudinal

and latitudinal gradients, watersheds, environmental changes and history of human

cases. A total of 12.079 sand flies were captured, Lu.neivai (85,6%), Lu.migonei

(11,6%), Lu.cortelezzii (2,6%) and Lu. shannoni, Lu.quinquefer y Brumptomyia spp.

(less than 0,1%). The first two species had epidemiological importance because are

vectors of Leishmania braziliensis. Human cases have not entered into the analysis

because they introduce errors by asymptomatic incidence and bias on spatial data

obtained from clinical files. In the analysis were used seven layers of BIOBLIM: annual

precipitation, maximum, minimum and seasonal temperature, and elevation. The

variables with the highest relative contribution were: seasonal rainfall, seasonal

temperature and elevation. This approach based on models gives us, once validated, the

ability to predict and assess the impact of enviromental changes on public health,

designing control strategies, and generate predictive models based on remote sensing

and captures of vectors in sentinel sites, in order to provide an early warning system for

outbreaks monitored in real time.

P-41

SANDFLY FAUNA (DIPTERA: PSYCHODIDAE) OF INTRA AND PERI -

DOMICILLIARY ENVIRONMENTS IN AGUAS CALIENTES, LA

CONVENCION, CUSCO, PERÚ.

QUISPE D(1)

, RADO D(1)

, QUISPE W(1)

, PACHECO R(1)

(1)Universidad Nacional de San Antonio Abad del Cusco

INTRODUCTION: Aguas Calientes is an endemic area for leishmaniosis in Perú, the

sandfly occurrence in intra and peri - domiciliary environments is studied here.

OBJECTIVE: To identify the Lutzomyia species in intra and peri - domiciliary areas in

Aguas Calientes, La Convención, Cusco, Perú.

METHODS: Six houses were visited to collect sandflies during august in 2007, CDC

light traps were used to collect the shadflies from 18:00 to 06:00 hours in intra and peri

- domiciliary environments, and Shannon traps with protected human bait from 18:00

to 22:00 hrs were used in peri - domicilary areas. The species identification was carried

out using Young and Duncan keys.

RESULTS AND CONCLUSIONS: Six species were collected in intradomicilliary

environments, these are: L. auraensis (36.36%), L. yuilli yuilli (18.18%), L. sherlocki

(18.18%), L. aragoi (9.09%), L. carrerai (9.09%) and L. walkeri. (9.09%). 11 species

were collected in peri - domiciliary environments: L. yuilli yuilli (60.28%), L. chagasi

(14.89%), and L. sherlocki (10.99%), were the most abundant species.

Lutzomyia yuilli was the most abundant species in the area, and L. auraensis was

typically intradomicilliar, the inhabitants of the area are highly exposed to the bites of

this species while are resting inside their houses.Acknowledgements: Anyela Gutierrez

and Karen Maguiña for their help during the study.

P-42

ABUNDANCIA RELATIVA DE ESPECIES DE LUTZOMYIA (DIPTERA:

PSYCHODIDAE) EN VEREDAS ENDÉMICAS PARA LTA, DEL NORESTE

COLOMBIANO

HOLVER PARADA 1, FAUSTINO MENDOZA

1, CARLOS ÁLVAREZ

2,

REINALDO GUTIÉRREZ1, ROCIO CÁRDENAS

1

1. Instituto de Investigaciones en Ciencias Biomédicas INBIOM de la Universidad

de Pamplona, Colombia. Telf. 0057-5682915 www.unipamplona.edu.co

2. Subgrupo Control de Vectores, Instituto Departamental de Salud Norte de

Santander, Colombia. Telf. 0057-5740753 www.ids.gov.co

Al Noroccidente de la Cordillera Oriental de los Andes suramericanos, se ubica el

Departamento de Norte de Santander y de sus cuarenta municipios 35 tienen

transmisión activa de Leishmanisis Tegumenaria Americana LTA. Según el Ministerio

de la Protección Social se ubicó entre los cinco primeros departamentos con las mayores

tasas de transmisión para LTA 30.4/100.000 hab en el país. Uno de sus municipios,

Abrego (bh-pM), con altura promedio de 1398 m.s.n.m y 21ºC, contiene veredas

endémicas para la transmisión de LTA y se mantiene en alto riesgo epidemiológico

desde el año 2003 según estadisticas del IDS. En este estudio de 13 veredas con

pesquisas entomológicas, 5 resultaron positivas para flebótomos: El Rosario, Los

Asientos, El Potrero, Mata de Fique y Anicillos. Consecutivamente durante 30 días, se

realizaron las recolecciones utilizando trampa Shannon y CDC, para un total de 2219

ejemplares capturados. Se identificaron 327 machos los cuales correspondieron con Lu.

longiflocosa, y 1892 hembras encontrando 3 especies: Lu. Longiflocosa (90.64%), Lu.

lichy y Lu. dubitans. Las veredas del Rosario (82.24%) y Los Asientos (17.21%)

presentaron las mayores abundancias. De los métodos de captura utilizados la trampa de

luz tipo Shannon fue la más efectiva con un porcentaje del 87.38%. Las mayores

capturas de Lu longiflocosa se llevaron a cabo en el peridomicilio de las viviendas, los

cuales son invadidos por cafetales y secundariamente por cultivos de cebolla y tabaco,

hallazgo que infiere la asociación indirecta de la población de flebótomos a este tipo de

vegetación. Pese a que en Norte de Santander se presentan 9 especies del Grupo

Verrucarum y se conoce la abundancia de Lu spinicrassa en los focos de mayor

endemicidad, es notoria la predominancia de Lu longiflocosa en esta extensa zona de

Abrego y su posible asociación con la ocurrencia de LTA en el municipio.

P-43

NEW RECORDS OF LUTZOMYIA FRANCA, 1924 (DIPTERA:

PSYCHODIDAE) IN THE ORINOCO REGION OF COLOMBIA.

RAFAEL J. VIVERO G1, LINA M. CARRILLO B

1, ANDRÉS VÉLEZ

1,

CAROLINA TORRES G1, SANDRA URIBE SOTO

2, EDUAR E. BEJARANO

3,

IVÁN D. VÉLEZ1

1. Research Associate, Program for the Study and Control of Tropical Diseases -

PECET, University of Antioquia. rajovigo2001@yahoo.com, Medellín, Colombia.

2. Molecular Systematics Research Group – National University 3. Biomedical

Research Group, University of Sucre, Sincelejo, Colombia

Introduction: The transmission cycles of parasites that cause leishmaniasis are defined

by the geographical distribution of insect vectors and by their preferences for specific

habitats. In ecoepidemiological studies, it is very important to identify the species

involved in transmitting the disease as well as their distribution, in order to determine

the risk of infection in different biogeographical areas.

Objective: To report the finding of seven species of Lutzomyia in an endemic area of

cutaneous leishmaniasis in the orinoquia region of Colombia.

Methods: The phlebotomine sand flies were collected in light traps such as CDC and

Shannon, from 18:00 to 22:00 hours, between November and December of 2007 in the

rural village of Caño Arriba, Vichada. The species were identified following Young and

Duncan (1994), and Galati (2003) taxonomic keys.

Results and Conclusions: Out of 98 individuals collected in traps there were nine

phlebotomine species, with seven species recorded here for the first time: L. saulensis,

L. flaviscutellata, L. runoides, L. yuilli, L. ayrozai, L. betinni and Lutzomyia sp.

(Evandromyia). The species L. aragaoi and L. antunesi had been reported in different

localities of the orinoquia, and it is worth to point out the presence of antropofilic

species like L. antunesi, which is considered a potential vector of Leishmania

guyanensis; L. yuilli found infected with L. panamensis; L. flaviscutellata, a vector of

Leishmania amazonensis and L. ayrozai, which is considered highly antropofilic and

associated to Leishmania naiffi. The presence of these especies in the peridomestic and

extradomestic environments is a potential risk for human transmission of leishmaniasis.

Financial Support: This research was supported by Secretary of Health's Department

Vichada.

P-44

SANDFLIES (DIPTERA: PSYCHODIDAE) IN THE ORIENTAL CHACO

PHYTOGEOGRAPHICAL REGION-ARGENTINA: SEASONAL AND

HABITAT DISTRIBUTION

SZELAG EA, ROSA JR, SALOMON OD1, ANDRADE FILHO JD

2, PEÇANHA

BRAZIL R3

Instituto de Medicina Regional. Universidad Nacional del Nordeste, Chaco, Argentina,

e-mail: szelag_enrique@yahoo.com.ar 1Ministry of Health/CONICET, Buenos Aires, Argentina.

2Instituto René Rachou, Fiocruz, Belo Horizonte, MG, Brasil.

3Instituto Oswaldo Cruz, Fiocruz, Rio de Janeiro, RJ, Brasil.

Chaco province is divided in two phytogeographical regions, Wet Chaco and Dry

Chaco, both with current Tegumentary Leishmaniasis (TL) cases. The aim of this work

was to identify the sandflies of the Dry region, almost unknown, and to determine their

seasonal and habitat distribution. Insects were collected in Misión Nueva Pompeya

(24º55’S, 61º30’W) from November 2006 to December 2007 in Fortín Arenales (FA)

and Los Pozos (LP), around houses with active LT cases. The captures were performed

monthly one night/site with CDC light traps from 19.00 to 07.00h in domestic,

peridomestic (animals shelters) and wooded habitats (≥150m from domicile). In total

1,689 sandflies were captured, with a sex ratio male/female 2.02: 1. The seasonal

distribution was: Spring: Lutzomyia migonei 80.1%, L cortelezzii complex 8.9%;

Autumn: L peresi 2.5%, L. quinquefer 0.8%, L. neivai 0.2%, L. spp 7.3%; Summer: L.

toroensis 0.2%; while none sandfly was obtained during winter. L. spp was sent to the

Centro de Referência Nacional e Internacional para Flebotomíneos of the Instituto René

Rachou/FIOCRUZ, Brazil, for further identification. There were not differences in

diversity between sites. In FA more phlebotomine were collected in the woods (36.1%)

than in the peridomestic (24.9%) or the domestic (23.1%) habitat. In LP there wasn't

differences between the domestic and woods captures (5.7% and 5.8%), but there are

difference with the peridomestic trapping (4.3%). L. migonei was prevalent in all

ecotones and seasons, followed by L cortelezzii complex. Based on the abundance and

closeness to houses with cases these species could be suspected as TL vectors in the

region

Financial Support: Roemmers Argentina Foundation, Fiocruz

P-45

SANDFLIES (DIPTERA: PSYCHODIDAE) OF THE CHACO PROVINCE,

ARGENTINA

ROSA JR1, SZELAG EA

1, SALOMON OD

2, ANDRADE FILHO JD

3,

CARVALHO GM3, STEIN M

1, PEÇANHA BRAZIL R

4

1Instituto de Medicina Regional. Universidad Nacional del Nordeste. Chaco, Argentina.

e-mail: juan_rosa05@yahoo.com.ar 2Ministry of Health/CONICET, Buenos Aires, Argentina.

3Instituto René Rachou, Fiocruz, Belo Horizonte, MG, Brasil.

4Instituto Oswaldo Cruz, Fiocruz, Rio de Janeiro, RJ, Brasil.

American Cutaneous Leishmaniasis (ACL) is endemic with occasional epidemics in

northern Argentina´s provinces, including Chaco. As the last Phlebotomine there were

reported before the`60s, this work aim was to update the sandfly diversity and

distribution in the Dry and Wet phytogeographical regions of Chaco province. From

2001 to 2007 in Dry Chaco [Misión Nueva Pompeya (24º55'S, 61º30W)] and Wet

Chaco [Margarita Belén (27º10`S,58º50`W) and Resistencia (27º26'S,58º54'W)] were

made regular captures twice a month with CDC light traps (19.00 to 07.00h) in

domestic, peridomestic and the forest. Eight species were captured: Lutzomyia neivai, L.

cortelezzii, L. shannoni, L. migonei, L. sallesi, L. quinquefer, L. peresi, L. toroensis and

Brumptomyia brumpti. The last six species were first records for Chaco province. L.

sordellii reported once in 1953 wasn’t found in this study. L. neivai prevailed in the Wet

Chaco in peridomicile and forest, but it was the less abundant in the dry region. L.

migonei reported as the second in abundance in most Argentinian regions, in Dry Chaco

was the first in all biotopes. L. cortelezzii, also suspected as vector in Argentina, was

captured in all ecotones together with L. sallesi. L. quinquefer, limited to the eastern

Paranaense region was found in the Wet Chaco in peridomestic and forest

environments. L. peresi was captured in houses and the forest in Dry Chaco and it was

the first report in Argentina and its southernmost record. L. toroensis was a new

American species, found in the Dry Chaco, in forest captures. In conclusion the sandfly

fauna of Chaco province is composed by nine species including L. sordellii historical

records. Based on their abundance and the literature the suspected vectors of ACL were

L. migonei and L. neivai in the Wet Chaco, and L. migonei and L. cortelezzii in the Dry

Chaco.

Financial Support: Roemmers Argentina Foundation and Fiocruz.

P-46

PHLEBOTOMINAE (DIPTERA: PSYCHODIDAE) IN AN ENDEMIC SITE OF

THE WET CHACO REGION, CHACO PROVINCE, ARGENTINA.

, 1

N OD², SOBRADO SV¹, SZELAG EA¹, STEIN M¹, SALOMOROSA JR

GORODNER JO¹

¹Instituto de Medicina Regional. Universidad Nacional del Nordeste. Resistencia,

juan_rosa05@yahoo.com.arChaco, Argentina. 2Ministry of Health/CONICET, Buenos Aires, Argentina.

During the last decades emergent outbreaks of American Cutaneous Leishmaniasis

(ACL) were reported in the phytogoegraphical region of Wet Chaco, mainly associated

with the gallery forests of the main rivers. With the aim to describe the sandfly fauna

and their seasonal variation in a site with ACL cases but low endemicity, we carried out

a study from November 2004 to October 2006 in Resistencia city (27º30’S, 59ºW).

Regular collections were performed twice a month in two sites (A-B) in domestic

(houses), and peridomestic (hen house and pigsty) habitats, and the forest, with CDC

light traps (19:00 to 07:00h.). A total of 1459 sandflies were captured with a sex ratio

M/F:1,98/1. Lutzomyia neivai (83%), L. migonei (14%), L. cortelezzii (2%), L. shannoni

(0,2%) and Brumptomyia brumpti (0,8%). Lutzomyia neivai and L. migonei

predominated in the peridomestic habitat and the forest in both sites. L. neivai was only

captured inside the house in site B, with two females. Phlebotomine were present all the

year round, except March, June, and July 2005, and September 2006. The seasonal

distribution (year 100%)-average temperature(ºC)-rainfall(mm) was: Winter 49.82%-

15.7ºC-8.3mm due to an exceptional temperate dry (mean temperature 27ºC), Spring

21.52%-22.8ºC-10mm, Summer 15%-27ºC-14.2mm, Autumn 13.63%-19.9ºC-22.2mm).

Mainly due to L. neivai prevalent species the captures showed a trimodal pattern that

peaks in April and November 2005, and July 2006. L. neivai and L. migonei were

dissected in search for natural infections of Leishmania, none was found, PCR is also

performed (ongoing results). Based on their abundance, distribution in the ecotone of

the forest and peridomestic habitats, and literature about epidemic 'hos spots' in the Wet

Chaco, we conclude that L. neivai and L. migonei could be also the probable vectors in

the region during inter-epidemic periods.

Financial Support: Roemmers Argentina Foundation

P-47

STUDIES ON THE VECTORS OF LEISHMANIASES IN THE STATE OF

TOCANTINS, BRAZIL: PORTO NACIONAL MUNICIPALITY,

TRANSMISSION AREA OF AMERICAN VISCERAL LEISHMANIASIS

MAURICIO L VILELA, BRUNO M CARVALHO, ELIZABETH F RANGEL

Lab.Transmissores Leishmanioses, Instituto Oswaldo Cruz/FIOCRUZ, Av.Brasil, 4365,

Rio de Janeiro, RJ, Brazil

The State of Tocantins, Brazil, has been presenting high levels of leishmaniases, of both

cutaneous (ACL) and visceral (AVL) forms. Despite this fact, available information

about the sand fly vectors is still scarce in the State. The objective of the present study

was to carry out an entomological survey in Porto Nacional, intense transmission area

of AVL, according to the Brazilian Ministry of Health. Sand fly captures were

conducted monthly using CDC light traps and Shannon traps, during 24 consecutive

months, from October 2004 to September 2006. A total of 4455 specimens were

collected and 38 species of the genera Brumptomyia and Lutzomyia were identified. The

Index of Species Abundance (ISA) was calculated in the rural and urban areas of the

municipality. On both areas, the highest ISA values were attributed to Lutzomyia

(Lutzomyia) longipalpis, vector of AVL (0.742 and 0.972 for rural and urban areas,

respectively). Compared to the results from the rural area, L. (L.) longipalpis showed a

higher distribution in the urban area. Lutzomyia (Nyssomyia) whitmani was more

representative in the rural area. Other potential sand fly vectors of ACL were identified:

L. (L.) gomezi, L. migonei, L. (N.) flaviscutellata, L. (N.) antunesi, and L. (N.)

intermedia. Despite the existence of some sand fly putative vectors of ACL, L. (N.)

whitmani is being suggested as the most important vector.

Financial Support: FIOCRUZ; CNPq-DECIT/2006;ENERPEIXE

IV: Molecular Methods for Detection of Leishmania in Sandflies

P-48

NATURAL INFECTION OF SANDFLIES (DIPTERA: PSYCHODIDAE:

PHLEBOTOMINAE) IN THE URBAN AREA OF CAMPO GRANDE, MS,

BRAZIL, BY Leishmania SPP.

ALESSANDRA GUTIERREZ DE OLIVEIRA1, BYANCA REGINA PAIVA

2,

ROSELY DOS SANTOS MALAFRONTE2, MARIA ELIZABETH

CAVALHEIROS DORVAL1, REGINALDO PEÇANHA BRAZIL

3, EUNICE

APARECIDA BIANCHI GALATI4

1. Universidade Federal de Mato Grosso do Sul, Laboratório de

Parasitologia/DPA/CCBS, Campo Grande, MS, Brazil, E-mail: agoliveira@nin.ufms.br.

2. Universidade de São Paulo, IMT, São Paulo, Brazil. 3. Instituto Oswaldo Cruz- LBII,

Rio de Janeiro, RJ, Brazil. 4. Universidade de São Paulo, FSP, Departamento de

Epidemiologia, São Paulo, SP, Brazil.

Natural infection of sandflies was investigated by the mini-exon PCR technique in

females caught weekly in three regions of remains of forest and five houses of Campo

Grande, MS. Of 563 females studied 103 were analyzed individually and 82 in pools of

2 and 10 specimens. Among 15 species studied the most frequent was Lutzomyia

longipalpis which reached 75,8% of the caught specimens. Positive results were found

in nine specimens of which four belonged to Lu. longipalpis species and the others to

Evandromyia lenti, Micropygomyia quinquefer, Nyssomyia whitmani, Psathyromyia

aragaoi and Psychodopygus claustrei, representing a minimum infection index of 1,6%.

The natural infection of sandflies by Leishmania spp is an important parameter for the

incrimination of species as vector of the parasite.

Financial support: FUNDECT 498/02, DECIT/FUNDECT, process 41/100208/2003

and FIOCRUZ.

P-49

DETECTION OF NATURAL INFECTION OF LUTZOMYIA LONGIPALPIS

(LUTZ & NEIVA, 1912) BY LEISHMANIA CHAGASI (CUNHA & CHAGAS,

1937) IN THE MUNICIPALITY OF JANAÚBA, STATE OF MINAS GERAIS,

BRAZIL

MICHALSKY E.M.1,3

, FRANÇA-SILVA J.C.2, BARATA R.A.

1, LOUREIRO,

A.M.F., TEIXEIRA-NETO4, R.G.

1; GUEDES K.S.

1, LARA-SILVA F.O.

1,

FORTES-DIAS C.L.5 & DIAS E.S.

1

1Centro de Pesquisas René Rachou/FIOCRUZ,

2 Universidade Federal de Minas Gerais,

3Universidade Federal do Triângulo Mineiro,

4Secretaria Municipal de Saúde de

Janaúba, 5Fundação Ezequiel Dias; Minas Gerais, Brazil.

Visceral leishmaniasis (VL) has been reported in almost every state in Brazil. Until the

1970’s, the transmission of VL was restricted to rural areas. Later on, a number of

human cases of VL have been recorded in urban areas of big, medium and small size

cities. Our study was developed in Janaúba, a city with 65,500 inhabitants in the north

of the Brazilian state of Minas Gerais, which has been considered as an area of intense

transmission of VL by the Brazilian Ministry of Health. A total of 126 human cases

were reported in the last five years and the prevalence rate of canine VL reached 17%.

The aim of our study was to determine the rate of L. longipalpis naturally infected by

Leishmania spp. and to identify the infecting species of the parasite in Janaúba.

Entomological captures were performed in 15 districts of Janaúba using HP light traps

and the phebotomine sand flies females were dissected for species identification. Pools

with 10 females of L. longipalpis each were subjected to genomic DNA extraction, PCR

amplification and RFLP analysis. All of the 155 pool samples analysed were positive

for the cacophony gene of Lutzomyia spp. thus validating the DNA extraction. The

120bp-fragment characteristics of Leishmania spp. was detected in 6 out of these 155

pool samples, which resulted in a natural rate of infection of 3.8% for L. longipalpis in

Janaúba. The RFLP patterns indicated Leishmania chagasi as the infecting parasite in

the municipality.

Financial support: CNPq, FAPEMIG, FIOCRUZ and UFTM

P-50

LUTZOMYIA ANTUNESI AS SUSPECTED VECTOR OF CUTANEOUS

LEISHMANIASIS IN THE ORINOQUIAN REGION OF COLOMBIA

ERIKA SANTAMARÍA1, ADOLFO VÁSQUEZ

2, ANGÉLICA GONZÁLEZ

2,

STELLA BUITRAGO3, AGUSTÍN GÓNGORA

2, OLGA CABRERA

1

1. Grupo de Entomología, Instituto Nacional de Salud. Bogotá D.C.

2. Universidad de los Llanos. Villavicencio, Colombia.

3. Secretaria de Salud del Meta. Villavicencio, Colombia.

Introduction: knowledge of cutaneous leishmaniasis epidemiology in the Orinoquian

region of Colombia is limited. In this region the disease is considered as predominant

of sylvatic areas, anyhow, in the last 5 years an increase of cases has been observed in

zones of agricultural and cattle expansion and even close to urban areas. In 2005, in a

rural zone few kilometers of Villavicencio city, were detected 15 cases of cutaneous

leishmaniasis in humans and six cases in canines. This outbreak motivated an

entomological study.

Objective: Identify the probable vectors of cutaneous leishmaniasis in the affected area

having into account the relative abundance of sand flies and their natural infection with

Leishmania spp.

Methods: Sampling of sand flies was done with CDC trap in and around 20 dwellings.

Pools of three females of the most abundant species of Lutzomyia were used to identify

Leishmania spp. by PCR, with OL1 y OL2 primers.

Results: 1,305 sand flies of nine species were captured, of which L. antunesi (75.5 %)

and L. walkeri (19.1 %) were the most abundant. Anthropophilic species L. panamensis

and L. gomezi were presented with low abundances (<2.4). The PCR detected

Leishmania spp. infection in two L. antunesi groups (total = 123 processed females).

Conclusion: Due to the fact that L. antunesi was the most abundant species and was

found with the Leishmania infection, it may be considered as the principal suspected

vector of cutaneous leishmaniasis in the study area. It is recommended to do detailed

studies on the biology of this species including bitting and resting behavior, in order to

carry on vectorial control measures.

P-51

FIRST REPORT OF LUTZOMYIA (NYSSOMYIA) NEIVAI (DIPTERA:

PSYCHODIDAE: PHEBOTOMINAE) NATURALLY INFECTED BY

LEISHMANIA (V.) BRAZILIENSIS IN A PERIURBAN AREA OF SOUTH

BRAZIL, STATE OF RIO GRANDE DO SUL, USING A PCR MULTIPLEX

ASSAY.

DANIELA PITA-PEREIRA1, GETULIO D SOUZA

2, ADRIANA ZWETSCH

3;

CARLOS ROBERTO ALVES1,

CONSTANÇA BRITTO1, ELIZABETH F.

RANGEL3

1Lab.Biologia Molecular e Doenças Endêmicas

3Lab. Transmissores de Leishmanioses,

Instituto Oswaldo Cruz, FIOCRUZ, Av.Brasil, 4365, Rio de Janeiro,RJ, Brasil 2Seção

de Reservatórios e Vetores, IPB, LACEN/RS, FEPPS, Secretaria Estadual de Saúde do

RS, Brasil

In order to identify Lutzomyia spp. naturally infected by Leishmania parasites, a PCR

multiplex coupled to non-isotopic hybridization was used for the analysis of wild sand

flies circulating in an endemic periurban area of American Cutaneous Leishmaniasis

(ACL) in the municipality of Porto Alegre (first human cases in 2002), State of Rio

Grande do Sul, Brazil. Sand flies were collected by CDC light traps from October 2006

to May 2007. A sample of 450 Lutzomyia (N.) neivai specimens was collected (270

females and 180 males) and grouped into pools of 10 insects each for DNA extraction

and Leishmania spp. detection by the proposed molecular assay. PCR positive results

were achieved in 3 out of the 27 Lutzomyia (N.) neivai female pools and the infection by

Leishmania (Viannia) braziliensis was confirmed after hybridization with a species-

specific biotinylated probe derived from the kinetoplast minicircle conserved sequence.

Considering the detection of three positive groups (3/27) and that there was at least one

infected insect in each, it was possible to infer an infection rate of 1.1% in the analyzed

samples. Our results in association with some ecological data suggest a potential

vectorial competence of Lu. (N.). neivai in transmitting L. (V.) braziliensis in the

municipality of Porto Alegre, south Brazil, and suggest this sand fly species as another

potential vector of L. (V.) braziliensis in Southeast and South Brazil.

Work support by: FIOCRUZ and CNPq, Brazil

P-52

INCRIMINATION OF LUTZOMYIA SPP. (DIPTERA: PSYCHODIDAE) AS

PROBABLE VECTOR OF LEISHMANIA IN AGUAS CALIENTES, LA

CONVENCION, CUSCO, PERU.

RADO D(1)

, QUISPE W(1)

, PEREZ JE(2)

, QUISPE M (1)

, VELAND N(2)

, ESPINOSA

D(2)

, PACHECO R(1)

(1) Universidad Nacional de San Antonio Abad del Cusco

(2) Instituto de Medicina Tropical Alexander von Humbolt, UPCH

Cusco is a region with one of the highest rates of leishmaniasis in Peru. Most of the

reported cases are of people who moved from Andean areas to the tropical forest areas

to work for short periods. Aguas Calientes, an endemic for Leishmaniosis, is one of

these areas, receiving workers for agriculture.

The Objective of this study was to incriminate Lutzomyia spp. as probable vector of

Leishmania in Aguas Calientes.

METHODS: Sandflies were collected with Shannon trap with protected human bait and

CDC light traps, in a dry and rainy season of 2005. To determine the natural infection of

Leishmania, we used three techniques; dissection of the sandflies guts, PCR and in-vitro

culturing of the isolated parasites. The isolated parasites from sandflies and recruited

patients were identified by PCR amplification, followed by PCR-RFLPs.

RESULTS & CONCLUSIONS: 15 species of the Lutzomyia genus and 1 of the

Brumptomyia genus were identified for Aguas Calientes. The most predominant species

were L. yuilli yuilli, L. llanosmartinsi, L. carrerai, L. hirsuta y L. chagasi; these species

were collected with the Shannon traps. Compared with previous reports, 5 species new

records were found for the study area, L. auraensis, L. walkeri, L. andersoni, L. choti

and Brumptomyia galindoi. Nine (9) natural infected samples were detected by the PCR

technique, 8 Leishmania positives belong to L yuilli yuilli and 1 to L. chagasi. The

presence of Leishmania brazilienzis “complex”, was detected by PCR-RFLP technique,

using the Hsp 70 gen. These parasites were isolated from 2 specimens of L. yuilli yuilli,

and also from parasites isolated from patients (both belonging at same study area) and

kept under in vitro-culture conditions. The results show L. yuilli yuilli as a probable

vector of Leishmania, because it was found naturally infected by the same Leishmania

species found in patients from de same area.

V: Sandflies as Hosts of Microorganisms other than Leishmania

P-53

STUDIES ON THE ACTIVITY PERFORMED BY SERRATIA MARCESCENS

IN PROMASTIGOTES OF LEISHMANIA (LEISHMANIA) INFANTUM

CHAGASI AND LEISHMANIA (LEISHMANIA) AMAZONENSIS IN VITRO AND

IN THE EXPERIMENTAL INFECTION OF LUTZOMYIA (LUTZOMYIA)

LONGIPALPIS .

SANDRA M P OLIVEIRA1, MARIANNA C COSTA

1, CHERYL GOUVEIA

1,

LUCIA H PINTO-DA-SILVA3, DEIVID C SOARES, ELVIRA M T SARAIVA

2 &

E F RANGEL1

1Laboratório de Transmissores de Leishmanioses, Instituto Oswaldo Cruz/FIOCRUZ,

2Departamento de Imunobiologia, IMPPG/UFRJ

Bacteria of genus Serratia are frequentely associated to insects of several orders and

some species was found in intestinal microbiota of sand fly, which could interfer on

development of promastigotes of Leishmania spp. Studies on the activity performed by

Serratia marcescens, strain SJ tio, on promastigotes of Leishmania (L.) infantum

chagasi and Leishmania (L.) amazonensis were carried out in vivo and in vitro. The role

of S. marcescens in the Lutzomyia longipalpis infection by L. (L.) infantum chagasi and

by L. (L.) amazonensis was evaluated through experimental infections of females fed

with blood containing: (1) promastigotes of L. (L.) infantum chagasi and L. (L.)

amazonensis (2.5 x 107/mL), (B) S. marcescens (1x10

8 and 1x10

6 CFU/mL), and (C)

promastigotes and bacteria. After the midguts dissection the co-infection was analyzed

by the optic microscope and the parasites were counted in Neubauer's chamber. The

agglutinated activity was tested using a suspension of S. marcescens (1x108

CFU/mL)

diluted in series, followed by the addiction of L. (L.) infantum chagasi and L. (L.)

amazonensis promastigotes (2.5 x 107/mL) and incubation in environmental

temperature. After the association with S. marcescens, in the concentrations 1x103,

1x104, 1x10

5 and 1x10

6 CFU/mL, the viability of L. (L.) infantum chagasi was analyzed

in cytometer. The bacteria didn’t demonstrate lysis effect on the promastigotes of both

Leishmania species, only the agglutinate effect was observed. However, in the co-

infected midguts, the promastigotes presented no motility and had their morphology

altered. The strain SJ tio was able to kill the L. longipalpis females, and also, lower the

infection degree. The results suggest that S. marcescens could have an important role as

a natural control of the leishmaniasis transmission process.

Financial support: FIOCRUZ; FAPERJ

VI: Ecology of Leishmaniosis

P-54

PARTICULAR CONDITIONS OF AMERICAN CUTANEOUS

LEISHMANIASIS TRANSMISSION IN AN OLD COLONIZATION AREA:

CAMPUS FIOCRUZ DA MATA ATLÂNTICA (ATLANTIC FOREST)

(JACAREPAGUÁ, RIO DE JANEIRO /RJ, BRAZIL).

CHERYL GOUVEIA1, ROSELY M. OLIVEIRA

2 & ELIZABETH F. RANGEL

1

1Lab. Transmissores de Leishmanioses, IOC/FIOCRUZ

2Deptº. Endemias Samuel Pessoa, ENSP/FIOCRUZ

The American Cutaneous Leishmaniasis (ACL) is characterized as a focal disease, with

ocurrence and maintenance features of transmission cycle very specific by locatily. This

study intends to know the social, economical and biological determinants related to the

ACL transmission in the Campus FIOCRUZ da Mata Altântica, represented by five

communities with about 700 inhabitants living in precarious houses nearly to the wood.

Quantitative and qualitative techniques were used to investigate the particular features

that could be associated to the occurrence and maintenance of the transmission cycle.

Was also done a monthly entomological investigation with light traps, during one year,

in three monitoring stations in the locality with the highest number of human cases of

ACL, Caminho da Cachoeira: sand fly collections were done in the intra and

peridomestic environments and at the wood limit. In the campus, the ACL occurs in

individuals of several ages, most between 0 to 10 years old, without gender and

occupation distinction. The abundance of Lutzomyia (N.) intermedia and L. migonei,

vectors of Leishmania (V.) braziliensis, in the intra and peridomestic environments, is

possibly related to the survival strategies developed by the popular groups, whose

maintain creations of domestic animals, giving conditions for breeding sites and shelters

to the vectors, besides acting as attractive for mammals sylvatic hosts. It was verified

that ACL occurrence is probably influenced by the way of life of the popular groups,

whose produces different conditions by receptivity for ACL. In this kind of situation,

proposes of environmental management and community mobilization become

important, since stimulating residents integration in order to create strategies avoiding

man’s exposition to the vector, without compromise the peculiar way of life of each

locality and without remove the survival strategies nescessary to the popular groups.

Financial Support: PDTSP/FIOCRUZ;CAPES

P-55

TREATMENT OF THE CUTANEOUS LEISHMANIASIS WITH

FITOTHERAPY IN MARGINAL-URBAN AND RURAL COMMUNITIES IN

TRUJILLO, VENEZUELA

ROJAS E.,CHÁVEZ O.,VIRLA J. & JUÁREZ E.

Postgrado en Educación para la Promoción de la Salud Comunitaria. Instituto

Experimental Jose Witremundo Torrealba NURR-ULA. AP 168

saludcomunitaria@ula.ve. Trujillo – Venezuela

The located cutaneous leishmaniasis is caused by the sting of a female phlebotomine

insect, previously infected by a parasite of the Leishmania genus. In Trujillo, Venezuela

this illness has high epidemic risk for its urban and rural character, it presents an index

of variation monthly average of 143 cases, calculated based on the voluntary attendance

of cases to the clinical center of this Institute since 1982.

We present the results obtained when contrasting two rural communities (group A) and

two marginal urban (group B) of this territory of the foot of Andean mountains with the

objective of knowing the personal practice in front of the perception of the

leishmaniasis as an illness. The sample consisted on 80 women and 20 men obtained by

reference of a previous work where they turned out to be responsible for the clinical

behavior of trying or not the wound(s) and of going or not to a hospital center for such

an end. Those selected consented by means of informed consent.

This type of study valorative descriptive demonstrated that beliefs persist and the

common practice in both groups of people (55 group A and 45 group B) about the use

of the Phytotherapy, differing alone in the form of obtaining the medicinal plants.

Regarding the application form the Leishmaniasis is treated with washes of infusions in

80% whose botanical species employees (15) have demonstrated to be able to antiseptic,

anti-inflammatory and healing. The uses of macerated and raw cataplasms for occulsive

cure of 10 different botanical species are effective for 40% of the interviewees in the

urban means and 60% of the rural means. The application of topical treatments with

dyes, oils and raw latex was effective in the first days of the illness.

75% of the two groups didn't go to medical establishments for that they obtained

clinical cure (scaring) in two or three months and the rest 15 urban% and 20 rural% they

needed local medical help for cure. 10% and 5% of them in each group received

immunotherapy.

The phytotherapy knowledge direct conserved traditionally by direct lineal familiar

relationships is very right as it demonstrates by the successful cure that was found. The

cost of the pharmaceutical medicines used universally for the leishmaniasis and the little

existence in the country, the restricted use of the antimonies makes that it is necessary

to re-evaluate the traditional outlines so that the chemical-specific treatment is applied

on time, don't be interfered and to avoid later complications without disqualifying the

use of the phytotherapy before and during the specific treatment..

P-56

EVALUATION OF VISCERAL LEISHMANIASIS VECTOR URBANIZATION

IN A CITY OF THE NORTH OF BRAZIL

, IORLANDO R. 1

S. DE OLIVEIRA, DAVI MARCOS 1

IVONEIDE M. SILVA

BARATA2, ADELSON A. A. SOUZA

2, EDILENE OLIVEIRA DA SILVA

1,

ELVIRA MARIA SARAIVA3

1Universidade Federal do Pará,

2Instituto Evandro Chagas,

3Universidade Federal do

Rio de Janeiro.

The number of visceral leishmaniasis cases has increased over the past ten years in

Brazil, especially in the north and northeast of the country. Visceral leishmaniasis is a

typically sylvan disease. However, environmental changes produced by human

intervention, such as deforestation, have produced the expansion of endemic areas and

urbanization of the disease and vector. The aim of this study was to evaluate the

visceral leishmaniasis vector urbanization in Barcarena (Pará), an endemic Visceral

Leishmaniasis area in north of Brazil. Sand flies were caught, using CDC light traps,

along an urban-rural gradient: a densely populated urban area, an intermediate, and a

rural area, located on the forest border, CDC traps were installed inside hen-house, 150

cm high. The sand flies caught were identified and the natural infection rate of females

were evaluated by dissection and microscopic examination. A total of 4.861 specimens

were collected (3.357 males and 1.504 females). Phlebotominae sand fly fauna was

comprised by nine species. The predominant specie was Lutzomyia longipalpis with a

rate of 96.75%, suggesting its participation in the transmission of Visceral

Leishmaniasis. A total of 1.382 L. longipalpis females were dissected and no

Leishmania infection was detected. All sand flies were caught at the border of the

forest, except for one, caught at the intermediate area. None sand flies was caught at

urban area suggesting that the transmission is still restricted to rural sites. However, the

fact that a specimen was collected in intermediate area indicates that the urbanization is

a real possibility and that the vector monitoring is important.

Grant: CNPq

P-57

IMPACT OF CLIMATE VARIABILITY ON AMERICAN CUTANEOUS

LEISHMANIASIS IN VENEZUELA

ALFONSO J. RODRIGUEZ-MORALES,1 LILIANA RADA,

2 JESUS

BENITEZ,1,3

CARLOS FRANCO-PAREDES,4

ELINA ROJAS.1

1Instituto Experimental Jose Witremundo Torrealba, Universidad de Los Andes,

Trujillo; 2Salud Miranda, Miranda;

3Ministry of Health, Maracay; Venezuela.

4Emory

University, Atlanta, USA.

Introduction and Objectives: American Cutaneous leishmaniasis (ACL), a tropical

vector-borne disease caused by Leishmania spp, vectorized in Americas by species

Lutzomyia, could be influenced by climatic variability. This issue has been understudied

in many places. For this reason we report possible impacts of climatic variability and El

Niño events occurred during 1994–2003 on ACL in 17 endemic states of Venezuela.

Methods: Climatic data was obtained from remote sensing systems. Epidemiological

data was obtained from Environmental health service (DGSACS), Ministry of Health.

Climatic events classification was made according NOAA and the indexes SOI and ONI

were used as main global climatic variability indicators. Comparisons of yearly

variations and deviation from medians trends between ACL incidence and climatic

variability as well lineal regression models were made. Statistical analyses were made

with SPSS 10.0 and GraphPad Prism 4.0, 95% of confidence.

Results: During this period a considerable global climatic change was present, with

strong El Niño events during years 1994, 1997 and 2002, and strong La Niña events

during 1995/1996 and 1998–2001. El Niño in eastern region of the country is expressed

as drought periods and La Niña as increases in rainfall. During this period, in these

states 17,589 cases of ACL were registered (20.2% from Lara, 11.9% Miranda and

10.5% Trujillo), mean of 2281.1±443.8 cases/year. During years with El Niño a mean

increase of 78.61% in ACL incidence was observed (ranging 8.3 to 614%), whilst in La

Niña a mean decrease of 9.78% was evidenced (ranging 8.3 to 614%). Comparisons in

deviation according these seasons were significant 15 out of 17 states (p<0.001). Lineal

regression models analysis, for Lara found that with a higher value of SOI (tending to

La Niña) less incidence of ACL is observed, although did not reached statistically

significance (r2=0.3085, p=0.0955); with higher values of ONI an increase in ACL

incidence was observed, being significant (r2=0.4254, p=0.0410). Similar patterns were

observed in other states.

Conclusions: This evaluated data reflected importance of climatic variability on ACL

incidence and phlebotomine sandflies vectorial transmission, and open further

investigations in the area to develop possible forecasting and monitoring systems with

relevance in regional public health.

VII: Sandfly Saliva

P-58

PHLEBOTOMUS PAPATASI SALIVARY GLAND SEQUENCE VARIABILITY

AND IMPACT ON DEFINING VACCINE CANDIDATES

RAMI MUKBEL1, MARCELO RAMALHO-ORTIGAO

1, VINITA TRIPATHI

1,

VALDIR BALBINO1, ILIANO COUTINHO-ABREU

1, GWEN STAYBACK

1,

MARIHA WADSWORTH1, EMIL LESHO

5,

EMAD DIN YEHIA FAWAZ2,

SHAABAN S. EL-HOSSARY2, HANAFI HANAFI

2, DAVID HOEL

2, MAHMOUD

ABO-SHEHDA3, SHADEN KAMHAWI

4, GLENN WORTMANN

5, AND MARY

ANN MCDOWELL1.

1University of Notre Dame, Notre Dame, IN, USA

2Naval Medical Research Unit #3, Cairo, Egypt

3Jordan University of Science and Technology, Irbid, Jordan

4LMVR, NIAID, NIH, Rockville, MD, USA

5Walter Reed Army Medical Center, Washington DC, USA

Data suggest that the incorporation of salivary components in multi-component

vaccines may be a viable strategy for the development of anti-Leishmania vaccines. If

sand fly saliva as a vaccine component is to be realized, it is necessary to understand the

variability of salivary genes and human immune responses to such variability. Here we

investigated expression and amino acid sequence variability of salivary gland proteins

from field populations of P. papatasi sand flies from the Middle East. Salivary gland

cDNAs encoding secreted proteins were PCR amplified, sequenced and the results were

compiled using various bioinformatics tools. For each protein, predicted MHC class II

T-cell epitopes were obtained and compared to areas of amino acid sequence variability.

Our results indicate greater sequence variability than was previously suggested and we

have identified additional MHC class II T-cell epitopes. To explore the early events of

anti-saliva human immune responses monocyte derived dendritic cells and macrophages

were generated from naïve individuals and the effect of sand fly salivary gland

homogenate (SGH) on co-stimulatory molecule expression and cytokine secretion was

assessed. Blood samples also were collected from US soldiers deployed to Iraq and

compared to US soldiers that have never been deployed to P. papatasi endemic regions.

Salivary antigens recognized by the sera of exposed individuals were identified by

western blot analysis of P. papatasi SGH. Peripheral blood mononuclear cells were

assessed for anti-SGH proliferative capacity and cytokine secretion.

P-59

IMMUNOGENEIC, PROTEOMIC AND MORPHOLOGY OF THE

SALIVARY GLAND OF LUTZOMYIA MIGONEI AND LUTZOMYIA

OVALLESI

NIEVES E., BUELVAS N., RONDON M., GONZALEZ N.

LAPEX-Laboratorio de Parasitología Experimental, Depto. Biología, Facultad de

Ciencias Universidad de Los Andes, Mérida, EDO-Mérida, Venezuela 5101

E-mail: nevelsa ula.ve

Salivary glands from Lutzomyia ovallesi (Ortiz) and Lutzomyia migonei (Franca)

(Diptera: Psychodidae), Leishmania vectors, were compared. Female sandflies groups

of both species were fed with 50% sucrose solution, blood meal and Leishmania

amazonensis infected blood. Protein concentration for females fed with 50% sucrose

solution was determined, for L. migonei an average of 0.8 µg/pair of glands and for L.

ovallesi an average of 0.5 µg/pair of glands. Salivary glands of L. ovallesi fed with 50%

sucrose solution and blood were smaller than those of L. migonei. This difference was

statistically significant (p<0.05). About 16-18 polypeptides were detected in L. ovallesi

and L. migonei from salivary glands with molecular weights ranging from 6 to 180 kDa.

Comparison between protein profiles by SDS PAGE of salivary glands from these two

species in different groups and post feeding timings showed that no apparent change in

the kinetic expression of salivary proteins in the different physiological states exists for

these two species exhibiting different qualitative and quantitative features. However, the

results showed proteins that are specific for L. migonei (25 and 32 kDa) and L. ovallesi

(15, 99, 100 and 170 kDa). Furthermore, polypeptides with high molecular weights in

both species, 170 and 180 kDa for L. ovallesi, and 180 kDa for L. migonei, were

detected. Also, antibodies against salivary gland specific proteins were found in mice

which were immunized by the bites from L. ovallesi (6, 14, 18, 20 and 29 kDa) and L.

migonei (6, 14, 20, 25 and 29 kDa), with 18 kDa and 25 kDA being specific for L.

ovallesi and L. migonei, respectively. Cross reactivity with the anti-saliva serum from

both species was observed; 4 polypeptides were recognized by L. ovallesi in anti-

migonei sera (6, 14, 25, and 29 kDa), and a polypeptide was recognized by the saliva of

L. migonei saliva against anti-ovallesi serum of 29 kDa. Results suggest that the sandfly

salivary proteins could be used as effective tools of screening risk factors in the

population and other vertebrate hosts, and as a potential candidates for the development

of a vaccine.

KEY WORDS: Lutzomyia, salivary glands, proteins, sandflies, Leishmania.

Financial Support: Project LOCTI and CDCHT-ULA.

VIII: Surveillance and Sandfly Control

P-60

PRELIMINARY OBSERVATIONS ON ATTRACTIVENESS OF

PHLEBOTOMINAE SAND FLIES (DIPTERA: PSYCHODIDAE) TO

DIFFERENT COLORED STICKY TRAPS.

BRAZIL RP1, BRAY DP

2, TEMELJKOVITCH M

3.

1- Instituto Oswaldo Cruz,Fiocruz,Rio de Janeiro,RJ,Brazil

2- Keele University, Staffordshire ,UK

3- Secretaria de Agricultura, Prefeitura de Corumbá, MS, Brazil.

Sticky traps, used extensively in agricultural pest management, could offer a cheaper

alternative to mechanical light traps for use in sand fly monitoring and control.

However, whether sand flies are attracted to such traps, and if they show any preference

for a particular colour in the field, is unknown. Here, we tested the relative

attractiveness of five colours of trap (green, black, red, yellow and blue, 16cm x 20cm)

coated with insect glue (Oecotack A6, Aerosol) in a pig pen in Corumbá, Mato Grosso

do Sul, Brazil. Trapping took place over four nights, with the position of each colour

rotated between replicates. Male and female Lutzomyia cruzi and Lutzomyia forattini

were captured, but numbers were generally low, with no significant effect of colour on

attractiveness (Kruskal-Wallis test). These results indicate that while sand flies can be

collected on sticky traps, use of a chemical attractant may be required to lure and kill

large numbers.

Financial Support: Faperj, Fiocruz and Wellcome Trust.

P-61

REEVALUATION OF THE FEEDING HABITS OF LUTZOMYIA

LONGIPALPIS IN THE CITY OF ARAÇATUBA, SÃO PAULO, BRAZIL,

FROM 2002 TO 2005, AFTER INTRODUCTION OF A DOG DELTAMETHRIN

COLLARS - SCALIBOR®.

VERA LF CAMARGO-NEVES¹, MARLUCI GUIRADO, LÍLIAN AC RODAS,

GISELE S CABRAL, CLÓVIS PAULIQUÉVIS JR.

Leishmaniasis Study Group, Surveillance Epidemiologic Center, Endemic Control

Superintendence. Secretary of Health of State São Paulo, Brazil.

¹E-mail: veracamargo@saude.sp.gov.br

A previous study in Araçatuba, São Paulo, Brazil, evaluated feeding habits of L

longipalpis in areas with transmission of visceral leishmaniasis, registering marked

preference for canine blood. We reevaluated feeding patterns of this species after

introduction of a deltamethrin collar, acknowledged repellent for the vector.

A cohort of dogs was performed during 2002 to 2005, in two areas of the city. In the

area called Collar, seronegative dogs used a collar impregnated with deltamethrin 4%-

Scalibor®

(from 2003 to 2004), and seropositive dogs were euthanized, according to the

guidelines of the official Program; in the Control area, infected dogs were euthanized.

Entomologic captures were made in each area, in an average of 40 households

each/month, with ideal characteristics for vector establishment, employing aspirators

(6V), within and without households per 20 minutes. Ingurgitated females were

separated and preserved at -22°C until intestinal contents exam, by immuneprecipitation

reaction, in capillary tube. Each female was immersed in 0.2ml of sterile physiological

solution at 0.85% overnight, and then macerated and centrifuged twice at 3000rpm/10’,

and afterwards tubes were filled with 30l of swimming fluid and 30l of total antisera

Sigma® human, dog, chicken, horse and mice. Reading time, determined from the

positive control for each antisera, varied from 12 to 24 hours. Proportion tests were

employed for analysis, with a significance level of 5%.

It was collected 13113 samples of L. longipalpis, 8455 (72.1%) were captured in the

Collar area and 89.2% were males. Of the 1420 females, 48.1% were ingurgitated; from

these 53.1% were examined with the anti-sera set, resulting in the Control area, 40.9%;

31.8%; 24.5%; 2.7% e 0.0% reactive, respectively, for dog, chicken, humans, horses

and mice and, in the Collar area, the percentages were 33.8; 31.1; 31.3; 0.5 e 3.2%,

respectively, for the same antisera.

We did not observe any statistically significant differences, during the period, in both

areas, and it was possible to see feeding preferences for blood from dogs, chickens and

humans, different that it was observed in the previous study, which can be explained by

the collection pattern employed in this study, directed to areas that favored the finding

of the vector. Introduction of the collar did not lead to changes in the feeding pattern of

the vector.

P-62

EFFECT OF FRUIT AND LEAVES OF MELIACEAE PLANTS

(AZADIRACHTA INDICA A. JUSS AND MELIA AZEDARACH L.) ON THE

DEVELOPMENT OF LUTZOMYIA LONGIPALPIS LARVAE (DIPTERA:

PSYCHODIDAE: PHLEBOTOMINAE), UNDER EXPERIMENTAL

CONDITIONS

CLAUDIA A. ANDRADE-COELHO1, NATALY A. SOUZA

1, CHERYL

GOUVEIA1, VANDERLEI C. SILVA

1, MARCELO S. GONZALES

2 &

1ELIZABETH F. RANGEL

1. Lab. Transmissores de Leishmanioses, Instituto Oswaldo Cruz/FIOCRUZ,Av. Brasil,

4365, Manguinhos, Rio de Janeiro, RJ 21045-900, Brasil. 2. Departamento de Biologia

Geral, Universidade Federal Fluminense,RJ,Brasil.

.

Some species of Meliaceae are known as potential sources of insecticidal substances.

Extracts from seeds and leaves of Azadirachta indica and Melia azedarach have shown

insecticidal properties against several medically relevant insects . Recent studies have

focused on the effects of azadirachtin, a triterpenoid originally extracted from Meliaceae

plants, on the development and mortality of Lutzomyia longipalpis larvae (Andrade-

Coelho 2006). The present laboratory study focuses on the feeding of L. longipalpis

larvae with unprocessed fruit and leaves of A. indica and M. azedarach in order to

determine the effects of these extracts on the mortality and metamorphosis of this sand

fly. Larvae were separated immediately after eclosion and placed in Petri dishes with

plaster of Paris as a substrate. At least three batches of 30 first-instar larvae each were

formed per study group. The larval food for the control groups were offered to the

larvae as daily dose. Experimental groups were fed with triturated fruit and leaves from

both Meliaceae plants and were offered to the larvae as daily dose.

A. indica and M. azedarach fruit and leaves significantly increased larval mortality in

comparison to untreated larvae fed. A. indica fruit and leaves and M. azedarach fruit

blocked the molting of the larvae to the fourth-instar. Feeding on M. azedarach leaves

resulted in stopped the development of larvae in the second-instar. No antifeedant effect

was detected. These data suggest that the non-toxic unprocessed extracts obtained from

A. indica and M. azedarach are potent development inhibitors of L. longipalpis larvae.

Financial Support: Fiocruz, CNPq and Faperj

P-63

THE SPREADING OF LEISHMANIASIS IN ITALY: ENTOMOLOGICAL

SURVEYS IN NORTHERN AND CENTRAL-WESTERN REGIONS

GRAMICCIA M., BONGIORNO G., DI MUCCIO T., GRADONI L., MAROLI,

M.

MIPI Department, Istituto Superiore di Sanità, Rome, Italy. E-mail:

marina.gramiccia@iss.it

Zoonotic Visceral Leishmaniasis (ZVL) incidence has been increased in Italy since

early 1990’s with new foci detected within traditional endemic foci but also in northern

regions previously recorded as non-endemic. From 2003 to 2005 a national project was

launched in 5 northern regions of Italy (Maroli et al 2008, Trop. Med. Int. Health, 13:

256-64) to monitor the ZVL putative spreading by human, canine and entomological

surveys. The surveillance continued in the frame of UE subproject EDEN-LEI.

Particularly, entomological surveys, aimed to study the distribution of putative sandfly

vectors in two selected north-western areas of sub-Alpine and sub-Apennine territories

bordering the Padana valley, were carried out since 2005. In 2007, a study was also

carried out in central-western Italy (Abruzzi region) with the aim to evaluate the

influence of altitude on the distribution of two Leishmania vectors, Phlebotomus

perniciosus and Phlebotomus perfliewi. Sandfly specimens were collected by sticky

traps settled in a variety of resting sites and identified to the species level. The

collecting site records were entered in a GIS database adopted by EDEN-LEI teams and

used to characterize the bio-geographic factors limiting the presence of phlebotomines.

In the sub-Alpine area two Leishmania vectors, P. perniciosus and P. neglectus, were

recorded from 206 to 570 m a.s.l; in the sub-Apennine area P. perniciosus and P.

perfiliewi were found from 52 to 440 m a.s.l. It was concluded that the most competent

L. infantum vector, P. perniciosus, is widespread in all the environments investigated.

On the contrary, the secondary vectors P. neglectus and P. perfiliewi show limited

distribution, being the first present only in the sub-Alpine area and the second in the

sub-Apennine one. Apparently, the large Padana valley seems to be free from

phlebotomine sandfly colonization. The sandfly survey carried out in central Italy

(Aquila and Teramo provinces of the Abruzzi region) revealed that among Phlebotomus

species, P. perfiliewi and P. perniciosus were the most abundant. Few specimens of P.

mascittii and one of P. neglectus were also collected. P. perniciosus, although less

abundant than P. perfiliewi, showed a constant prevalence and was recorded up to 772

m a.s.l.. Instead, P. perfiliewi was the prevalent species in the range 300-599 m a.s.l.

(50.2%) but it was apparently absent from 610 m a.s.l.

This study was carried out in the framework of 6° F.R.P Integrated Project Emerging

Diseases in changing European eNvironment (EDEN), CE N. 010284-2, Subproject

Leishmaniasis.

P-64

EVALUATION OF INCANDESCENT OR UV LIGHT TRAPS FOR SANDFLIES

CATCHES IN ARACAJU – SE, BRAZIL

JERALDO, V.L.S.;1 CASANOVA, C

2; MELO, C.M.

1; ARAÚJO, E.D.

1; CRUZ,

D.E.R.1; GÓES, M.A.O.

1; PINTO, M.C.

3

1Instituto de Tecnologia e Pesquisa, Aracaju-SE;

2Superintendência de Controle de

Endemias, Mogi Guaçu-SP;

3Faculdade de Ciências Farmacêuticas, UNESP,

Araraquara-SP

The monitoring of sandflies is a crucial steep in programmes for control of

leishmaniasis. The knowledge of the sandflies species involved in a transmission area as

well as their population dynamics can guide measures of reduction of the disease

transmission. In August of 2007 sand flies catches were carried out in Areia Branca a

rural area of Aracaju/SE, Brazil. The aims of this study were to identify the local

sandfly fauna and to compare a modified CDC light traps with incandescent or UV

lights. Such comparison was used to define which it would be the better source of light

for collecting sandflies in a future more extensive survey in the area. During six nights,

two nights a week, the traps with incandescent light and UV were disposed in pairs with

three to five meters of distance apart each other in six different sites. The traps were

shifted of position each second night of the week. A total of 72 traps (12 traps/night

and 36 with each source of light) were used. The Mann-Whitney statistical test was used

to analyze the results. The sandflies species collected were Lutzomyia longipalpis (146),

Lutzomyia lenti (9) e Lutzomyia intermedia (1). The highest captures of L. longipalpis

were in UV traps, (U= 318,00, p=0,0002 for males and U= 404,50 p=0,006 for

females). Interestingly females with eggs were only collected in UV traps (13 specimens). The highest

effectiveness of UV suggest its possible use in monitoring L. longipalpis when its density is low.

P-65

THE BG-SENTINELTM

TRAP AS A TOOL TO CATCH PHLEBOTOMINE

SAND FLIES (DIPTERA: PSYCHODIDAE)

, 2

, ADELSON A. A. SOUZA2

, IORLANDO R. BARATA1

IVONEIDE M. SILVA

MARA C. PINTO3, CAROLINA SPIEGEL

4, ANDREY J. ANDRADE

5,

CLAUDIA C. GULIAS GOMES5, ÁLVARO E. EIRAS

5

1Universidade Federal do Pará,

2Instituto Evandro Chagas,

3Universidade Estadual

Paulista, 4Universidade Federal Fluminense,

5Universidade Federal de Minas Gerais.

Leishmaniasis is a wide problem of Health Public in Brazil. The diseases control is based

on treatment of human cases, euthanize of positive dogs and the monitoring of the sand

flies is carried out by CDC light traps. The BG-SentinelTM

(BGS) is a new trap that uses

a synthetic human odor as attractant (BG-LureTM

). The BG-Lure was developed to

attract the mosquito Aedes aegypti but attracts also other haematophagous insects. The

aim of this study was to evaluate the effectiveness of BGS to catch sand flies. Four

parameters using the BGS were evaluated: light source, trap height, opening position and

lure (BG-Lure).. Three experiments were carried out in peridomestic sites in Barcarena

(Pará, Brazil), an endemic Visceral Leishmaniasis area (1) to compare CDC and BGS

(without light, UV and incandescent lights; (2) to evaluate the BGS opening (insect

suction tube) in three different positions (up, down and lateral, at 40 and 150 cm high);

(c) to evaluate the BGS-lure with the best light source, height and opening position

(BGS+lure, BGS+light, BGS+lure+light). The CDC light trap was used as a control in

all field bioassay (latin square design). The traps collected a wide diversity of sand flies

species, including Lutzomyia longipalpis. The BGS trap was not efficient in the capture

sand flies without light source. Incandescent light was more attractive than UV light,

although statistical difference was not detected. CDC trap caught more sand flies than

BGS with incandescent (10.94±3.71; 3.18±1.21) as well as UV light (7.53±1.78;

2.47±0.74). The highest catches were observed when BGS-Sentinel was positioned with

the opening down at 150 cm high. BG-lure did not increase the capture of sand flies,

with or without the presence of light. The data suggest that the BGS-Sentinel is able to

catch sand flies but is not a substitute to CDC light trap.

Grants: FINEP; CNPq

P-66

LARVICIDAL ACTIVITY OF ESSENCIAL OIL PIPER ADUNCUM L. GAINST

LUTZOMYIA LONGIPALPIS (DIPTERA:PSYCHODIDAE)

JUNIOR, A. O. A; FILHO M. B. S; JUSTINIANO, S.C. B.

Centro Universitário Nilton Lins, Manaus, Amazonas, Brasil.

sjustiniano@niltonlins.br

Lutzomyia longipalpis is the most important vector of AmericanVisceral Leishmaniasis

(AVL) due to Leishmania chagasi in the New World. In an effort to find efective and

affordable ways to control this sand fly, the larvicidal activities of essencial oil of the

Piper aduncum plant was analyzed. 150 larvae of 3rd

larval instars were used for the

experiment, oil was diluted with etila acetate in three concentrations (10-1

, 10-2

and 10-3

)

and each dilution had three repetitions with 3µm. 10 larvae were put of in rearing pots

of 60mL, in a period of 72 hours. In this test they were made three replicates for each

concentration accompanied with the positive control (SPT® insecticide) and

negative (acetate). The pots were maintained at 27oC and 92% relative humidity

(RH) and light:dark cycle of LD 12:12. In 72 hrs, the experiment of dilution 10-1

obtained a percentage of 42% of larvae died, the dilution 10-2

obtained 22% of larvae

died and the dilution 10-3

obtained 11% of died, already the positive control obtained

20% of died and the negative control obtained 5% of died. The preliminary results

suggest a potential right of action on the larvae of Lutzomyia longipalpis, however new

tests should be accomplished to obtain the ideal concentration of that oil as larvicidal.

Financial support: CNPq ; FAPEAM.

P -67

PERCEPTION OF THE LEISHMANIASIS IN MARGINAL URBAN AND

RURAL COMMUNITIES IN TRUJILLO-VENEZUELA.

HERNÁNDEZ, MARÍA, DELGADO M, ALDANA A, MORÓN C, BRICEÑO Y,

CABRERA M. BENCOMO M. VIRLA J, ROJAS E.

Postgrado en Salud Comunitaria. Instituto Experimental JWT-NURR-ULA. AP168

saludcomunitaria@ula.ve. Trujillo-Venezuela.

The tegumentary American leishmaniasis is a known as tropical and subtropical illness

with importance in Public Health to be transmitted by phlebotomines vectors of

Leishmania both with autochthonous genres and species.

To exist the illnesses they should be recognized as social nosological entities, each

cultural group ends up having different perceptions. The objective of this work was to

establish the perception of Leishmaniasis, known illness as rural, of marginal urban

inhabitants of two communities (Las Adjuntas and Santa Rosa) and two rural (Loma de

Piedras Negras and Sabanetas). Previous validation study, they established focal groups.

A questionnaire type knowledge, practices and aptitudes was applied. It was carried out

a retrospective clinical study based on IDR and presence of corporal scars and medical

histories.

People of both groups (160) identified Leishmaniasis due to open lesions and closed in

skin or for scars though this illness is endemic for almost 450 years in these towns.

The biggest difficulty was in the recognition of the phlebotomine and of the causal

agent of the lesions mainly in the marginal urban communities. The lesion is recognized

but it doesn't exist an idea of how it is acquired.

The result of the personal practices was heterogeneous in both groups being the mothers

in 80% and the fathers 20% responsible for the decision about which medical or

homemade treatment. We conclude that the increase of cases of cutaneous leishmaniasis

in the urban area has contributed to the recognition of the lesion but it exist some

disagreements among knowledge and the personal practices adopted especially in front

the cases of self-medication in the marginal urban areas and the phytotherapy in the

rural area. The human beings live in societies and we create cultures.

P-68

GEOGRAPHICAL ATLAS OF THE CUTANEOUS LEISHMANIASIS IN THE

TRUJILLO STATE – VENEZUELA

CAÑIZALES B & ROJAS E.

Postgrado en Salud Comunitaria. Instituto Experimental JWT-NURR-ULA. AP 168

saludcomunitaria@ula.ve. Trujillo-Venezuela

At the present time a new paper of the Geography assumed as interdisciplinary and

auxiliary to the medical sciences for the development of technologies emerges that,

applied from teaching centers they allow to know the geographical origin and its

relationship with vulnerable areas for the diagnosis and control of some tropical

illnesses, as well as the eco-epidemiology description of the landscape where they

happen. The fundamental purpose of the present work was to elaborate a Geographical

Atlas of the Trujillo State; with base to the permanency and localization of clinical cases

of Cutaneous Leishmaniasis; to be used as didactic tool in the teaching of social and

health related areas in the primary schools of the Trujillo state - Venezuela, the type and

design of the investigation was documental, accompanied by an electronic prototype of

the Geographical Atlas of the Leishmaniasis in the Trujillo state. The investigation was

descriptive and transversal, when considering the clinical cases of Cutaneous

Leishmaniasis reported by the Experimental Institute of the Núcleo Universitario Rafael

Rangel, of the University of Los Andes, in the period understood among 2002-2005.

The determination of the geographical conditions of the municipalities that integrate the

Trujillo state, as well as the location of clinical cases of the mentioned pathology and

the permanency of the same ones, they were decisive elements in the design of the

Geographical Atlas. It was found that the municipality of Trujillo is the one that has

reported the bigger number of clinical cases of this illness in the analyzed period,

followed by Pampanito and Pampan. In relation to the geographical conditions, the

municipalities of this state present characteristic of always green vegetation, placed in

the categories of Tropical Humid Forest, Mount Foot Forest, among others. In relation

to the relief, the municipalities in those that the presence of cutaneous leishmaniasis is

perceived are located in valleys, fact that, together to the presence of cultivations of

coffee as the main agricultural activity, as well as to the temperature, they constitute

factors that favor the presence of the transmitter vector of the pathology before

mentioned, assigning to this the possibilities of acquiring the endemic character.

ANNEX 1

VISIT TO THE RIMAC VALLEY October 29,2008 Map showing our route.

HIGHLIGHTS OF THE VISIT

The Verrugas bridge or Carrion's bridge was built around 1870; the name was given as

a memorial of more than 7000 workers who died with Carrion's disease. Many of these

workers were foreign people hired for the construction of the rail way.

Tornamesa train station, at 1430 masl built in 1865-1870; this is the original building,

where CHT Townsend collected sandflies for the first time in Peru, from a window of

the building. He described Phlebotomus verrucarm Townsend 1913 with such material.

San Jeronimo de Surco, at 2000 masl a small town in an endemic area. Many

inhabitants have uta scars in their faces. Those interested can capture Lutzomyia in the

town surroundings.

AUTHORS INDEX

ABBASI I. OP-26

ABO-SHEHDA M. OP-41, P-58

ACARDI SA. OP-21, OP-22

AFONSO MMS. P-04, P-15

AKOPYANTS N. OP-34

ALDANA A. P-67

ALENCAR RB. P-20

ALKAN MZ. OP-27

ALMEIDA LDGD. OP-06

ALTEN B. OP-02, OP-29

ALVAREZ C. P-42

ALVES CR. P-51

ALVES VR. P-29

AMBROSIO VO. P-30

AMIRIH M. OP-46

ANDRADE AJ. P-65

ANDRADE AJD. P-15

ANDRADE ARO. OP-24

ANDRADE-COELHO CA. P-19, P-62

ANDRADE FILHO JD. OP-06, OP-11,

OP-25, OP-32, P-01, P-02, P-05,

P-06, P-11, P-31, P-32, P-37, P-45,

P-44

ANNAJAR B. P-29

ANTONIOU M. OP-02

AÑEZ N. OP-23

AQUINO RB. OP-24

ARAKI AS. OP-01, P-14

ARANA M. OP-42

ARBIZU G. OP-14

ARAUJO ED. P-64

ARAUJO IB. OP-01, P-14

ARRUDA CCP. OP-24

AZAVEDO ACR. P-28, P-38

BABUAZDE G. OP-44

BALBINO V. OP-41, P-58

BALCIOGLU C. OP-27

BARBOSA SBL. P-32

BARATA IR. P-56, P-65

BARATA RA. P-18, P-30, P-36, P-49

BARRESI G. OP-10

BARRETT TV. P-20

BASTOS MC. P-30

BAUZER LGSR. OP-01, P-14

BEATI L. OP-04, OP-17, P-08

BEJARANO EE. P-42

BENCOMO M. P-67

BENITES J. P-57

BEVERLY S. OP-34

BITTON S. OP-09

BONGIORNO G. P-63

BRAHIM LRN. P-38

BRAY DP. P-60

BRAZIL RP. OP-01, OP-06, OP-11,

P-01, P-06, P-14, P-17, P-26, P-27,

P-31, P-37, P-44, P-45, P-48, P-60

BRICEÑO Y. P-67

BRITTO C. P-51

BUELVAS R. P-59

BUITRAGO S. P-50

CABALLERO NNG. P-17

CABRAL GS. P-61

CABRERA OL. P-39, P.50

CACERES A. OP-17, P-08

CADENA H. P-34

CAMARGO-NEVES VLF. P-61

CAMPINO L. OP-35

CAMPOS M. OP-42

CANESE A. P-17

CAÑIZALES B. P-68

CABRERA M. P-67

CARDENAS R. P-42

CARDONA LAA. OP-08

CARRILLO LM. OP-08, P-43

CARVALHO BM. P-37, P-47

CARVALHO DA. OP-25, OP-32

CARVALHO GMDL. OP-06, OP-11,

P-02, P-05, P-06, P-11, P-37, P-45

CASANOVA C. P-64

CHANTURIA G. OP-44

CHAVEZ O. P-55

COHNSTAEDT LW. OP-17, P-08

COLEMAN RE. OP-28, OP-52

CONTRERAS MA. P-09

COSTA MC. P-53

COSTA SM. OP-03, P-21, P-28, P-38

COSTA WA. P-21, P-38

COSTA-PINTO D. P-07

COUTINHO-ABREU I. OP-41, P-58

CRISANTE G. OP-23

CRUZ C. OP-18

CRUZ CFR. P-23, P-24, P-25

CRUZ DER. P-64

CRUZ HC. P-28

CUNIO R. OP-26

DANTAS-TORRES F. OP-05

DAS P. OP-49

DA SILVA EO. P-56

DA SILVA FILHO OF. P-07

DAVIES CR. OP-47

DE ANDRADE AJ. OP-05, P-01

DE ANDRADE MR. P-01

DE OLIVEIRA AG. P-26, P-27, P-48

DE OLIVEIRA DMS. P-56

DE OLIVEIRA EF. P-25

DELGADO M. P-67

DEMIR S. OP-27

DEPAQUIT J. OP-13

DERSCHUM H. OP-46

DIAS ES. P-18, P-30, P-36, P-49

DI MUCCIO T. P-63

DISTEFANO S. OP-10

DO CARMO FF. P-01

DOLIDZE N. OP-44

DORVAL MEC. P-25, P-27, P-48

DOS SANTOS CB. P-02

DOS SANTOS F. P-11

DRAHOTA J. OP-40

DUARTE R. P-15

D’URSO V. OP-10

DVORAK VIT. OP-02

EIRAS AE. P-01, P-16, P-65

EL-ABAIED. P-29

EL-BUNI A. P-29

EL-HOSSARY SS. OP-41, P-58

ELNAIEM DA. OP-39

ERTABAKLAR H. OP-27

ESPINDOLA IAC. P-27

ESPINOSA D. P-52

ESPINOSA YR. P-35

FAIMAN R. OP-26

FALCAO AL. OP-25, OP-32, P-05,

P-32, P-37

FAULDE MK. OP-28, OP-46

FAWAZ EDY. OP-41, P-58

FERNANDES CE. P-27

FERNANDEZ M. OP-18

FERNANDEZ N. P-35

FERNANDEZ R. P-03, P-22

FERREIRA GEM. OP-01, P-14

FERRO C. P-08, P-33, P-34, P-39

FLORES M. OP-18

FLORES R. OP-51

FORDELLONE-CRUZ MF. P-23,

P-24, P-25

FORTES-DIAS CL. P-49

FRANCA-SILVA JC. P-18, P-36, P-49

FRANCO-PAREDES C. P-57

GALATI EAB. OP-05, OP-24, P-23,

P-24, P-25, P-27, P-48

GIORGOBIANI E. OP-44

GOES MAO. P-64

COMES CCG. P-65

GOMES CR. OP-25

GONGORA A. P-50

GONTIJO CMF. OP-32

GONZALES MS. P-62

GONZALEZ A. P-50

GONZALEZ N. P-59

GORODNER JO. P-46

GOUVEIA C.P-53, P-54, P-62

GRADONI L. P-63

GRAJAUSKAS AM. P-37

GRAMICCIA M. P-63

GUEDES KS. P-49

GUTIERREZ R. P-42

HAMILTON JGC. OP-01, P-14, P-17

HANAFI H. OP-41, P-58

HERNANDEZ J. OP-18

HERNANDEZ M. P-67

HEYL G. OP-46

HOCHBERG LP. OP-28

HOEL D. OP-41, P-58

HOLTHERM H-U. OP-46

HOSTOMSKA J. OP-40, OP-43

ILANGO K. OP-07

IVOVIC V. OP-02

JAFFE CL. OP-26

JECNA L. OP-35, OP-37

JERALDO VLS. P-64

JONES LA. OP-04

JUAREZ E. P-55

JUSTINIANO SCB. P-12, P-66

KAMHAWI S. OP-41, OP-44, P-58

KHALFA Z. OP-31

KIMBLIN N. OP-34

KIRSCH P. OP-53

KOZLOWSKY D. P-38

KRUEGER A. OP-46

KUMAR N. OP49

LANTOVA L. OP-30

LANZARO GC. OP-39

LARA-SILVA FO. P-36, P-49

LAWYER P. OP-34, OP-44

LEE JS. OP-28

LEGER N. OP-13

LESHO E. OP-41, P-58

LEUTENEGGER C. OP-39

LINS RMM. P-10

LLANOS-CUENTAS A. OP-19

LISI O. OP-10

LIZARRALDE DG MS. P-40

LOPES MOG. P-30

LOPEZ E. P-22

LOPEZ V. P-03

LOROSA ES. P-18

LOUREIRO AMF. P-18, P-49

MAASSEN W. OP-46

MACHADO TO. OP-06

MAGUIÑA-VARGAS C. OP-20

MAIA C. OP-35

MAIA JF. P-12

MALAFRONTE RDS. P-48

MALLET J. P-21

MARIN D. P-33, P-34

MARIN RG. P-35. P-59

MARLUCI G. P-61

MAROLI M. OP-02, OP-10, P-63

MASON PW. OP-36

MATZNER O. OP-50

MCAVIN JC. OP-28, OP-45

MCDOWELL A. OP-41, P-58

MEDINA V. OP-23

MELO CM. P-64

MELO MN. OP-25, OP-32

MENDOZA F. P-42

MENESES CRV. OP-03, OP-39

MICHALSKY EM. P-18, P-30, P-36,

P-49

MILLER D. OP-51

MIRANDA JC. P-04, P-15

MOCK S. OP-04

MORON C. P-67

MORWINSKY T. OP-46

MOSQUERA L. P-389

MOTTA-SILVA D. P-07, P-38

MOULTON JK. OP-28

MUKBEL R. OP-41, P-58

MUNDAL K. P-22

MUNSTERMANN LE. OP-04, OP-17,

P-08, P-33, P-34

NASEREDDIN A. OP-26

NIEVES E. P-59

NUNES VLB. OP-24

OCAMPO C. P-33, P-34

O`GUINN ML. OP-28

OGUSUKU E. OP-18, OP-42, OP-54,

P-13

OLGEN K. OP-27

OLIVEIRA RCA. P-30

OLIVEIRA RM. P-54

OLIVEIRA SMP. OP-38, P-53

ORSHAM L. OP-09, OP-31, OP-50

OVALLOS P-35

OZBEL Y. OP-27

PACHAS P. OP-16

PACHECO R. P-41, P-52

PAIVA BR. P-48

PARADA H. P-42

PARANHOS FILHO AC. P-26

PARDO RH. OP-47

PASSOS W L. P-31

PATRICK R. OP-34

PAULA IRP. P-23, P-25

PAULIQUEVIS JR C. P-61

PAZ GF. P-30

PEIXOTO AA. OP-01, OP-38, P-10,

P-14, P-19

PEREZ JE. OP-14, OP-15, OP-42,

OP-54, P-13, P-52

PINHEIRO AC. P-30

PINTO MC. P-62, P-28, P-64, P-65

PINTO IDS. P-02

PINTO-DA-SILVA LH. P-53

PITA-PEREIRA D. P-51

PITALUGA NA. OP-36

PUTMAN JL. OP-28

QUINTANA MG. OP-21, OP-22, P-40

QUISPE D. P-41

QUISPE M. P-52

QUISPE W. OP-54, P-41, P-52

RADA L. P-57

RADO D. OP-54, P-13, P-41, P-52

RAJAN A. OP-49

RAMALHO-ORTIGAO M. OP-41,

P-58

RAMIREZ O. OP-42

RANGEL EF. OP-03, P-04, P-07, P-15,

P-16, P-20, P-28, P-38, P-47, P-51,

P-53, P-54, P-62

RAWTON E. OP-52, OP-53

REQUENA E. P-03

REZENDE CF. OP-25

ROCCA MEG. OP-24

RODAS LAC. P-61

RODRIGUEZ-MORALES A. P-35,

P-57

ROHOUSOVA I. OP-40, OP-43

ROJAS A. OP-23

ROJAS E. P-55, P-57, P-67, P-68

RONDON M. P-59

ROSA JR. OP-21, OP-22, P-44, P-46,

P-45

ROWTON E. OP-51

RUBIN DE CELIS M. OP-14

RUBIO AL. P-09

SACKS D. OP-34, OP-44

SADLOVA J. OP-33, OP-35

SALOMON OD. OP-21, OP-22, P-40,

P-43, P-45, P-46

SANDOVAL CM. P-35

SANGUINETTE CDC. OP-06, OP-11,

OP-25, P-05, P-06, P-11, P-32

SANTAMARIA E. P-39, P-450

SANTOS JMM. P-12

SANTOS MFC. OP-24

SARAIVA EMT. P-53, P-56

SARAIVA L. OP-25, OP-32

SCHNUR L. OP-26

SCHRADER J. OP-46

SECUNDINO N. OP-34

SERRAO JE. P-21

SHEM KM. OP-08

SIDDIQUI NA. OP-49

SILVA EAE. P-26

SILVA IM. P-56, P-5

SILVA VC. P-19, P-62

SIMSEK F. OP-27

SOAREZ DC. P-53

SOARES LF. P-36

SOARES MJ. OP-38

SOBRADO SV. P-46

SOLORZANO N. P-22

SOONG L. OP-39

SOTO SU P-09, P-43

SOUSA LC. P-04, P-15

SOUZA AAA. P-56, P-65

SOUZA CM. OP-25

SOUZA GD. P-51

SOUZA JL. P-28

SOUZA NA. OP-01, P-10, P-14, P-19,

P-62

SPIEGEL CN. OP-38, P-65

STANCIL J. P-03

STAYBACK G. OP-41, P-58

STEIN M. P-45, P-46

SVAROVSKA A. OP-37

SVOBODOVA M. OP-26, OP-29,

OP-30

SWABY CJA. OP-45

SWANSON KI. OP-28

SZEKELY D. OP-09

SZELAG EA. P-44, P-44, P-46

TAVARES MG. OP-03, P-04

TEDESQUI V. P-31

TEMELJKOVITCH M. P-31, P-60

TEIXEIRA-NETO RG. P-49

TIVIROLI VA. P-26

TORRES G C. P-09, P-43

TORRES K. OP-42

TOZ SO. OP-27

TRAUB-CSEKO YM. OP-36

TRIPATHI V. OP-41, P-58

VALINSKY L.OP-31

VARGAS E. OP-23

VASQUEZ A. P-50

VELAND N. P-52

VELEZ A. OP-08, P-43

VELEZ ID. OP-08, P-09, P-43

VERGARA D. OP-08

VERMA RB. OP-49

VIDAL J. P-28

VIEIRA AP- OP-24

VIGODER FM. OP-01, P-14

VILELA ML. P-38, P-47

VIRLA J. P-55

VIVERO R. OP-09

VIVERO RJ. P-09, P-43

VLKOVA M. OP-43

VIRLA J. P-67

VOLF P- OP-02, OP-26, OP-29, OP-30,

OP-33, OP-35, OP-37, OP-40,

OP-43

VOLFOVA V. OP-43

VOTYPKA J. OP-26, OP-29, OP-30,

OP-40

VOTYPKA VIT. OP-02

WADSWORTH M. OP-41, P-58

WARBURG A. OP-26

WARD RD. P-19

WASSERBERG G. OP-12, OP-51,

OP-53

WHITE G. OP-48

WORTMAN G. OP-41, P-58

YÉPEZ JY. OP-23

ZOLLNER GE. OP-52

ZWETSCH A. P-51

SPONSORS

Vicerrectorado de Investigacion (UPCH)