40 Aniversario

6th INTERNATIONAL SYMPOSIUM

on PHLEBOTOMINE SANDFLIES

October 27-31, 2008, Lima, PERU

40 Aniversario

6th INTERNATIONAL SYMPOSIUM

on PHLEBOTOMINE SANDFLIES

October 27-31, 2008, Lima, PERU

Auditorio Hugo Lumbreras Cruz, Casa Honorio Delgado
Av. Armendriz 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 Arvalo
Ciro Maguia
Humberto Guerra
Elena Ogusuku
Carmen Rado
Pablo Villaseca
Dionicia Gamboa
HONORARY COMMITTEE
N. Aez, 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. Lger, 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:
Lunch:
Afternoon:

09:00-13:00
13:00-14:00
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 Maguia
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).
1

ALEJANDRA S. ARAKI , FELIPE M. VIGODER1, LUIZ G. S. R. BAUZER ,

GABRIEL E. M. FERREIRA1, NATALY A. SOUZA2, IZENEIDE B. ARAJO3,
JOHN G. C. HAMILTON4, REGINALDO P. BRAZIL5, ALEXANDRE A.
PEIXOTO1
1

Laboratrio de Biologia Molecular de Insetos, IOC, Fundao Oswaldo Cruz, Rio de

Janeiro, Brazil. 2Laboratrio de Transmissores de Leishmanioses, IOC, Fundao
Oswaldo Cruz, Rio de Janeiro, Brazil. 3Curso de Cincias Biolgicas e Agrrias,
Universidade Estadual do Piau, Parnaba, Piau, Brazil. 4Centre for Applied
Entomology and Parasitology, Institute of Science & Technology in Medicine, Keele
University, United Kingdom. 5Laboratrio de Bioqumica e Fisiologia de Insetos, IOC,
Fundao 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 VOTYPKAa, VLADIMIR IVOVICb, MARIA ANTONIOU c,
MICHELE MAROLI d, BULENT ALTEN e and PETR VOLFa
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 COSTA, CLAUDIO RV MENESES, MARA G TAVARES3 &
ELIZABETH F RANGEL
Lab. Transmissores de Leishmanioses, Instituto Oswaldo Cruz/FIOCRUZ; University
of California, Davis, Department of Pathology, Microbiology and Immunology 3Dep.de
Biologia Geral, Universidade Federal de Viosa,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 Ilhus (BA) (type locality), Londrina (PR), Santarm 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 MOCK1, LYNN A. JONES2, AND LEONARD E.
MUNSTERMANN2
1
2

Georgia Southern University, Statesboro, Georgia, USA

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, Galatis
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 JOS DE ANDRADE1, FILIPE DANTAS-TORRES2 & EUNICE
APARECIDA BIANCHI GALATI3
1

Departamento de Parasitologia, Universidade Federal de Minas Gerais, Belo

Horizonte, MG, Brasil - bioandrey@gmail.com.br
2
Departamento de Imunologia, Centro de Pesquisas Aggeu Magalhes, Fundao
Oswaldo Cruz, Recife, PE, Brasil - fdt@cpqam.fiocruz.br
3
Departamento de Epidemiologia, Faculdade de Sade Pblica, So 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:
Esprito Santo, Minas Gerais, Rio de Janeiro, and So 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 So Paulo (67), Rio de Janeiro (58), and Eprito
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.
JOS DILERMANDO ANDRADE FILHO1, TMARA OLIVEIRA MACHADO2,
LUSY DISNEY GOMES DE ANDRADE ALMEIDA3, GUSTAVO MAYR DE
LIMA CARVALHO1, CRISTIANI DE CASTILHO SANGUINETTE1,
REGINALDO PEANHA BRAZIL4
1- Lababoratrio de leishmanioses - Centro de Pesquisas Ren Rachou - Fiocruz
2-Setor de Controle das Leishmanioses, Malria e Raiva- Secretaria Municipal de Sade
CCZ - Palmas - TO
3 -Laboratrio de Entomologia - Secretaria Municipal de Sade- CCZ - Palmas - TO
4- Laboratrio de bioqumica 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 Esperana Farm (1022.921 S and 4803.587 W, 504 m.a.s.l.) in the
Taquaruu 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 Referncia Nacional e Internacional para Flebotomneos (CRNIF) of the Instituto
Ren 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 Piau, 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 MONDRAGN SHEM1, LUZ ADRIANA
ACOSTA CARDONA1, RAFAEL VIVERO1, DANIELA VERGARA1, ANDRS
VLEZ1, IVN DARO VLEZ1
1

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

Antioquia, Medelln, Colombia
2
Facultad de Ciencias Agrarias de la Universidad de Antioquia, Medelln, 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 BITTON2, DAVID SZEKELY3
1

Laboratory of Entomology, Ministry of Health, Jerusalem, Israel.

Regional Municipality Mate Binyamin, Psagot, Mizrah Binyamin, Israel.
3
Ministry of Environmental Protection, Jerusalem, Israel.
2

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 & THEODORS
INVESTIGATIONS ON MEDITERRANEAN KALA AZAR (1931)
V. DURSO1, O. LISI1, S. DISTEFANO1, G. BARRESI1, M. MAROLI2
1

Department of Animal Biology M. La Greca, University of Catania, Italy; 2MIPI

Department, Istituto Superiore di Sanit, 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 subunits (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 & Theodors observations.

OP-11
SANDFLIES
(DIPTERA:
PSYCHODIDAE)
CAPTURED
IN
THE
SPELEOLOGICAL PROVINCE OF BAMBU, DISTRICT OF MEDIUM AND
HIGH SO FRANCISCO, MINAS GERAIS, BRAZIL
GUSTAVO MAYR DE LIMA CARVALHO1, CRISTIANI DE CASTILHO
SANGUINETTE1, REGINALDO PEANHA BRAZIL2, JOS DILERMANDO
ANDRADE FILHO1
1- Laboratrio de Leishmanioses - Centro de Pesquisas Ren Rachou - Fiocruz
2- Laboratrio de Bioqumica 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 Bambu, 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
1

Walter 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 recolonization 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 recolonization 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. LGER 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 malesfemales 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 Per 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, Uiga 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
DIVERSITY OF PERU.

BARTONELLOSIS

IN

THE

GEOGRAPHICAL

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 BEATI4,
AND LEONARD E. MUNSTERMANN3
1

Seccin de Entomologa, Instituto de Medicina Tropical Daniel A. Carrin, Facultad

de Medicina Humana, Universidad Nacional Mayor de San Marcos, Lima, Per
2
Laboratorio de Entomologa, Instituto Nacional de Salud, Lima, Per
3
Yale University School of Public Health, New Haven, Connecticut, USA
4
Georgia 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. Direccin 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 Martn, Amazonas, Hanuco 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 MAGUIA-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 Nio 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
SALOMN ODa, QUINTANA MGb, ROSA JRc, ACARDI SAd
a

: Ministry of Health/CONICET, b: National University of Tucumn (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 80s 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
SALOMN ODa, ACARDI SAb, ROSA JRc, QUINTANA MGd
a

: Ministry of Health/CONICET, b: National University of Misiones (UNaM),

c
: National University of NorthEast (UNNE), d: National University of Tucumn
(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 Asuncin
(Paraguay), phlebotomine surveillance was intensified in the northern border of
Argentina. Lu. longipalpis was found in Clorinda (Formosa province) on the border
with Asuncin 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
NSTOR AEZ1*, ERSY VARGAS2, AGUSTINA
MEDINA2, GLADYS CRISANTE1, JOS Y. YPEZ2

ROJAS1,

VICENTE

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

Mrida, 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 G2005000370 (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. NUNES2, E. A. B. GALATI3, A. R. O. ANDRADE1, A.
P. VIEIRA2, M. E. G. ROCCA4, R. B. AQUINO4, C. C. P. ARRUDA1.
1

Centro de Cincias Biolgicas e da Sade, UFMS, Campo Grande, Brazil.

Centro de Cincias Biolgicas, Agrrias e da Sade, UNIDERP, Campo Grande,
Brazil. 3Faculdade de Sade Pblica, USP, So Paulo, Brazil,
4
Ncleo de Controle de Zoonoses, Prefeitura do Municpio de Bonito, Bonito, Brazil.
2

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 5210 S; 56o
3506 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 CARVALHO2, CRISTIANI
CASTILHO SANGUINETTE2, CAMILA RAGONEZI GOMES1, CARINA
MARGONARI SOUZA2, CHRISTIAN FREITAS REZENDE1, JOS
DILERMANDO ANDRADE FILHO2, ALDA LIMA FALCO2, 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 FAIMAN1, IBRAHIM ABBASI1, ABEDELMAJEED
NASEREDDIN1,
LIONEL
SCHNUR1,
RUBEN
CUNIO1,
MILENA
2
2
2
1
SVOBODOVA , JAN VOTYPKA , PETR VOLF , CHARLES L. JAFFE
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 20012003, 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 Shean, 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
MAPPNG OF RSK AREAS FOR VSCERAL AND CUTANEOUS
LESHMANASS RELATED WTH DSTRBUTON OF VECTOR SPECES
N WESTERN PART OF TURKEY USNG GEOGRAPHC INFORMATON
SYSTEMS
YUSUF OZBEL1, I. CNEYT BALCIOGLU2, KRAM LGEN3, FATH
SIMSEK4, SERAY OZENSOY TOZ1, HATCE ERTABAKLAR5, SAMYE
DEMIR6, M. ZYA ALKAN1
1

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

Celal Bayar University Medical School Department of Parasitology, Manisa, Turkey
3
Ege University Faculty of Letters Department of Geography, Bornova, Izmir, Turkey
4
Adnan Menderes University Science Faculty Department of Biology, Aydin, Turkey
5
Adnan Menderes University Medical School Department of Parasitology, Aydin,
Turkey
6
Mustafa Kemal University Science Faculty Department of Biology, Hatay, Turkey
2

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. OGUINN,3 & MICHAEL K. FAULDE5
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 complexparasites 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 VOTPKA, MILENA SVOBODOV, BULENT ALTENa, PETR VOLF
Department of Parasitology, Faculty of Science, Charles University, Prague, Czech
Republic, e-mail: vapid@natur.cuni.cz
a
Department 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 Ehrlichs 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, nonconserved 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 CARVALHO2, CLIA MARIA
FERREIRA GONTIJO2, JOS DILERMANDO ANDRADE FILHO2, ALDA
LIMA FALCO2, MARIA NORMA MELO1.
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 2m thick and consists of
thin laminar outer layer and thick amorphous inner layer. The thickness of PM
increased to about 5m 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 AKOPYANTS2, NICOLA KIMBLIN1,
RACHEL PATRICK1, NAGILA SECUNDINO1, STEPHEN BEVERLEY2, 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 Fundao Cincia 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 YM1.
1 - Laboratrio de Biologia Molecular de Tripanosomatdeos e Flebotomneos, 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 betagalactosidase, diminished replication of the VLP-encoded genome.
Conclusions: These results are the first indication for a nucleic acid-induced, nonspecific 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. OLIVEIRA2, ALEXANDRE A.
PEIXOTO2 & MAURILIO J. SOARES3
1

Universidade 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. MENESES2, C. LEUTENEGGER2, L. SOONG3 & 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 vectors 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 flyexposed 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-ORTIGAO1, VINITA TRIPATHI1,
VALDIR BALBINO1, ILIANO COUTINHO-ABREU1, GWEN STAYBACK1,
MARIHA WADSWORTH1, EMIL LESHO5, EMAD DIN YEHIA FAWAZ2,
SHAABAN S. EL-HOSSARY2, HANAFI HANAFI2, DAVID HOEL2, MAHMOUD
ABO-SHEHDA3, SHADEN KAMHAWI4, GLENN WORTMANN5, AND MARY
ANN MCDOWELL1.
1

University of Notre Dame, Notre Dame, IN, USA

Naval Medical Research Unit #3, Cairo, Egypt
3
Jordan University of Science and Technology, Irbid, Jordan
4
LMVR, NIAID, NIH, Rockville, MD, USA
5
Walter Reed Army Medical Center, Washington DC, USA
2

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 nave 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 ARANA2,
OSWALDO RAMIREZ2, MIGUEL CAMPOS2 & ELENA OGUSUKU3
1

Instituto de Medicina Tropical Alexander von Humboldt, Universidad Peruana

Cayetano Heredia. kike@upch.edu.pe
2
Facultad de Ciencias y Filosofa, UPCH.
3
Direccin 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 (Huarochir, 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
Investigacin, 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 vectorborne 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
1
2

National Center for Disease Control, Republic of Georgia;

National 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
90s 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
OPERATIONS WITH DEPLOYABLE, REAL-TIME
DISEASE AGENT ANALYTIC CAPABILITY

SURVEILLANCE
VECTOR-BORNE

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, fieldsustainable, real-time, arthropod-borne disease agent detection platform used to support
disease surveillance operations at far forward locations. The VSAS operates as a standalone 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 realtime 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 MORWINSKY2, HENRI DERSCHUM3,
MOHAMMED
AMIRIH4,
WINFRIED
MAASSEN2,
HANS-ULRICH
HOLTHERM2, GERHARD HEYL2, JOACHIM SCHRADER2, 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 DAVIES2
1

Universidad de La Salle, Bogot, 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 subandean 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
MATERIALS FOR CONTROLLING PHLEBOTOMINE SAND FLIES

AND

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) Patna800 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 casefatality 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 peoples 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 Kalaazar, 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 cooperation at community level.

OP-50
PERSPECTIVES OF SAND FLY CONTROL IN THE JUDEAN DESERT
LAOR ORSHAN1 AND ORNA MATZNER2
1
2

Laboratory of Entomology, Ministry of Health, Jerusalem

Pest 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 HOSTTARGETED INSECTICIDE.
GIDEON WASSERBERG1, DAVID MILLER2, EDGAR RAWTON1
1
2

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

Genesis 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,
RUSSELL E. COLEMAN.

ROXANNE

FLORES,

EDGAR

ROWTON,

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 (-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 -cypermethrin LLIN caused higher feeding inhibition.
Surprisingly, of a total 60 sand flies that were able to squeeze through -cypermethrinor 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 KIRSCH2, GIDEON WASSERBERG1
1
2

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

APTIVE 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 sandflies. 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 plantderived 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-flys 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:
(Addition).

Ecology, Distribution, and Population Dynamics of Sandflies

OP-54

SANDFLY FAUNA OF CENTRO DE INVESTIGACION Y CAPACITACION

RIO LOS AMIGOS (CICRA), MADRE DE DIOS, PERU.
PEREZ1, J. ENRIQUE; DIANA RADO2, WILFREDO QUISPE2 & 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, Per.
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 Per, 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 castaa 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 JOS DE ANDRADE1, FELIPE FONSECA DO CARMO1, MATEUS
RAMOS DE ANDRADE1, LVARO EDUARDO EIRAS1, REGINALDO
PEANHA BRAZIL2 & JOS DILERMANDO ANDRADE-FILHO3
1

Departamento de Parasitologia, Universidade Federal de Minas Gerais, Belo

Horizonte, MG, Brasil - bioandrey@gmail.com
2
Instituto Oswaldo Cruz, Fundao Oswaldo Cruz, Rio de Janeiro, RJ, Brasil
3
Centro de Pesquisas Ren Rachou, Fundao 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
Gacha, 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 Gacha:
(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 ESPRITO SANTO, SOUTHEAST
BRAZIL
JOS DILERMANDO ANDRADE FILHO1, ISRAEL DE SOUZA PINTO2,
CLAUDINEY BIRAL DOS SANTOS2 & GUSTAVO MAYR DE LIMA
CARVALHO1
1 - Laboratrio de Leishmanioses, Centro de Pesquisas Ren Rachou/Fiocruz
2 - Unidade de Medicina Tropical, Universidade Federal do Esprito 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 Esprito Santo, in the Reserva Biolgica 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 &
Falco, 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. FERNNDEZ1, V. LPEZ1, E. REQUENA 2AND J. STANCIL1
1
2

United States Naval Medical Research Center Detachment, Lima, Peru.

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 TAVARES2, JOS C MIRANDA3,
LINDEMBERGH C SOUSA4 & ELIZABETH F RANGEL1
1

Lab. 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 Mdica Dr. Thomaz Correa Arago, SESAUCE, 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, Jequi/BA, Sobral and Massap/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 Jequi and Teresina populations produced an individual set from
those formed by the Cear populations. In population of Jequi, when compared to
population of Cear, it can be observed that 79% to 95% of the characters revealed
significant differences. By ANOVA, at the crossroads between the populations of
Cear, few differences were detected. The dendogram created by UPGMA method
revealed a split between the Cear 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 Cear and Teresina/Jequi 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 FALCO,
CRISTIANI DE CASTILHO SANGUINETTE, JOS DILERMANDO
ANDRADE FILHO
Laboratrio de Leishmanioses - Centro de Pesquisas Ren 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
Referncia Nacional e Internacional para Flebotomneos of the Instituto Ren
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 & Falco, 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 PEANHA
BRAZIL2, CRISTIANI DE CASTILHO SANGUINETTE1, JOS DILERMANDO
ANDRADE FILHO1
1- Laboratrio de Leishmanioses - Instituto Ren Rachou - Fiocruz
2- Laboratrio deBioqumica 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
(Galvo & 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.
DANIEL MOTTA-SILVA, DIAMAR COSTA-PINTO, OCTAVIO
FERNANDES DA SILVA FILHO, ELIZABETH FERREIRA RANGEL
Lab. Transmissores de Leishmanioses, Lab. Epidemiologia Molecular de Doenas
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, Ilhus (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 Ilhus 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 BEATI2, ABRAHAM G. CACERES3,
CHRISTINA FERRO4,AND LEONARD E. MUNSTERMANN1
1

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

Georgia Southern University, Statesboro, Georgia, USA
3
Universidad Nacional Mayor de San Marcos, Lima Peru
4
Instituto Nacional de Salud, Bogota, Colombia
2

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 subgenusevansi, 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.
longiflocosaP. sauroida and P. torvidaP. 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 wellsupportedphylogenetic 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, MARA A. CONTRERAS1, CAROLINA TORRES G1,
ANDRS LPEZ RUBIO2, SANDRA URIBE SOTO2 , IVN D. VLEZ1 .
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, Medelln, 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 speciesspecific.
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. SOUZA2 AND A. A. PEIXOTO1
1

Laboratrio de Biologia Molecular de Insetos, IOC, Fundao Oswaldo Cruz, Rio de

Janeiro, Brazil. 2 Laboratrio de Transmissores de Leishmanioses, IOC, Fundao
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, Cear 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, FTIMA DOS SANTOS2,
GUSTAVO MAYR DE LIMA CARVALHO1, JOS DILERMANDO ANDRADE
FILHO1
1- Laboratrio de Leishmanioses - Centro de Pesquisas Ren Rachou - Fiocruz
2- Fundao Nacional de Sade, 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,
Rondnia, popularly known as Parque Ecolgico, 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 Ecolgico of Porto Velho. The new species from
Rondnia 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 applelike 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
PSYCHODIDAE) POPULATIONS OF CENTRAL AMAZON

(DIPTERA:

JUSTINIANO, SCB1, MAIA, JF2 ; SANTOS, JMM2.

1

Centro Universitrio Nilton Lins, Manaus, Amazonas, Brasil

nstituto Nacional de Pesquisas da Amaznia, Coordenao de Pesquisas em Cincias
da Sade, Manaus, Amazonas, Brasil.
justiniano@niltonlins.br
2

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 Airo, 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 RADO2 & ELENA OGUSUKU3
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 C, 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).
1

ALEJANDRA S. ARAKI , FELIPE M. VIGODER1, LUIZ G. S. R. BAUZER ,

GABRIEL E. M. FERREIRA1, NATALY A. SOUZA2, IZENEIDE B. ARAJO3,
JOHN G. C. HAMILTON4, REGINALDO P. BRAZIL5, ALEXANDRE A.
PEIXOTO1
1

Laboratrio de Biologia Molecular de Insetos, IOC, Fundao Oswaldo Cruz, Rio de

Janeiro, Brazil. 2Laboratrio de Transmissores de Leishmanioses, IOC, Fundao
Oswaldo Cruz, Rio de Janeiro, Brazil. 3Curso de Cincias Biolgicas e Agrrias,
Universidade Estadual do Piau, Parnaba, Piau, Brazil. 4Centre for Applied
Entomology and Parasitology, Institute of Science & Technology in Medicine, Keele
University, United Kingdom. 5Laboratrio de Bioqumica e Fisiologia de Insetos, IOC,
Fundao 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 AFONSO, ROSEMERE DUARTE2, JOS C MIRANDA3,
LINDEMBERGH C SOUSA4 & ELIZABETH F RANGEL
Lab. Transmissores de Leishmanioses, IOC/FIOCRUZ; 2Laboratrio de Pesquisa e
Servios em Sade Pblica, ENSP/FIOCRUZ; 3Laboratrio de Imunoparasitologia,
CPqGM/FIOCRUZ; 4Laboratrio de Entomologia Mdica Dr. Thomaz Correa Arago,
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 Jequi (BA), Teresina (PI), Sobral (CE) and Massap (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
Jequi and Massap. 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 Jequi,
Sobral and Massap. 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 mammals 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 JOS DE ANDRADE1, ELIZABETH F.RANGEL2, LVARO
EDUARDO EIRAS1
1

Depart. de Parasitologia, Universidade Federal de Minas Gerais, Belo Horizonte,

MG,bioandrey@gmail.com/alvaro@icb.ufmg.br
2
Lab.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 hydrocarbons characterization, using the complex L.
whitmani s.l. as model. Sand flies were caught in the five geographical regions from
Brazil: North (Par State); Northeast (Cear and Bahia States), Southeast (Minas Gerais
State), South (Paran 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 ASUNCIN, PARAGUAY.
BRAZIL RP1, HAMILTON JGC2, CABALLERO NNG1, CANESE A3.
1

- Lab. de Bioqumica 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 Mdicas, Universidad Nac. de Asuncin, 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 onespot (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)-9methylgermacrene-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 Asuncin, 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 JANABA, STATE
OF MINAS GERAIS, BRAZIL.
MICHALSKY E.M.1,3, FRANA-SILVA J.C.2, LOROSA E.S.4, LOUREIRO
A.M.F.5, BARATA R.A.1, DIAS E.S.1
1

Centro de Pesquisas Ren Rachou/FIOCRUZ-MG, 2Universidade Federal de Minas

Gerais-MG, 3Universidade Federal do Tringulo Mineiro-MG, 4Instituto Oswaldo
Cruz/FIOCRUZ-RJ, 5Secretaria Municipal de Sade de Janaba; 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 Janaba, 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. WARD & ALEXANDRE A. PEIXOTO
3

Lab. Transmissores de Leishmanioses & 2 Lab. Biologia Molecular de Insetos,

Instituto Oswaldo Cruz/ Fiocruz, Rio de Janeiro, Brazil; Institute of Science and
Technology in Medicine, School of Life Sciences, Keele University, Staffordshire, U.K.
3

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, Cear. 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 bursttype 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 Amaznia, 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, 3JOS EDUARDO SERRO
1

Depart. Biologia Animal, Universidade Federal De Viosa-Mg, 2Lab. Transmissores de

Leishmanioses, IOC/ Fiocruz, RJ; 3Departamento de Biologia Geral, Universidade
Federal de Viosa-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 FERNNDEZ1, NELSON SOLORZANO2, ETTY LOPEZ3 & KIRK
MUNDAL1
1 Naval Medical Research Center Detachment, Lima-Peru. (NMRCD).
2 Ministerio de Salud. Direccin Regional de Salud de Ancash. Hospital de Caraz.
3 Ministerio de Salud. Direccin Regional de Salud de San Martn.

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
Carrions 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 PARAN 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 Paran. 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 (230200 S and
50 2200 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 vectors 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 SO FRANCISCO, STATE OF PARAN,
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 Paran, 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 So Francisco is situated between the
cities of Cornlio Procpio 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 Paran.
Financial Support: FUNDACIN 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 PARAN, BRAZIL, 2005 -2007.
FORDELLONE-CRUZ, MF*,GALATI, EAB**,CRUZ, CFR* PAULAIR*/
*UENP/FALM-PR **FSP/USP-SP
INTRODUCTION: The Vila Velha Park in the Paran 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 Paran.
FINANCIAL SUPPORT: FUNDACIN 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 FALCO DE OLIVEIRA1, ANTONIO CONCEIO PARANHOS
FILHO2, ELAINE ARAJO E SILVA3, MARIA ELIZABETH CAVALHEIROS
DORVAL1, REGINALDO PEANHA BRAZIL4, VALTER ALEXANDRE
TIVIROLI2, ALESSANDRA GUTIERREZ DE OLIVEIRA1.
1. Universidade Federal de Mato Grosso do Sul - Laboratrio de
Parasitologia/DPA/CCBS, Campo Grande, MS, Brazil. 2. Universidade Federal de
Mato Grosso do Sul - Laboratrio de Geoprocessamento para Aplicaes
Ambientais/DHT/CCET, Campo Grande, MS, Brazil 3. Centro de Controle de
Zoonoses, Secretaria de Sade do Municpio 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 PEANHA
BRAZIL2, ITALO ALEXANDER CABELLO ESPINDOLA3, MARIA
ELIZABETH CAVALHEIROS DORVAL1, CARLOS EURICO FERNANDES1,
EUNICE APARECIDA BIANCHI GALATI4.
1. Universidade Federal de Mato Grosso do Sul, Laboratrio 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. Acadmico do curso de
Farmcia/Universidade Catlica Dom Bosco-UCDB, Campo Grande, MS, Brazil. 4.
Universidade de So Paulo, FSP, Departamento de Epidemiologia, So 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 Sade 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 ES1, OLIVEIRA RCA1, MICHALSKY EM1, PINHEIRO
AC2, AMBRSIO VO2, LOPES MOG3, PAZ GF1, & BARATA RA1.
1

Centro de Pesquisas Ren Rachou/FIOCRUZ; 2. Secretaria Municipal de Sade 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 CORUMB, BRAZIL.
BRAZIL RP1, LANA PASSOS W1, TEMELJKOVITCH M2, TEDESQUI V1,
ANDRADE FILHO JD3
1

- Lab. de Bioqumica e Fisiologia de Insetos - Instituto Oswaldo Cruz, Rio de Janeiro,

Brazil
e-mail: rpbrazil@ioc.fiocruz.br
2Secretaria de Agricultura-Prefeitura de Corumb, MS,Brazil.
3
Instituto Ren 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
4964; W 580 3695) 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 Corumb, 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 CEAR, BRAZIL?
JOS DILERMANDO ANDRADE FILHO, SOFIA BERNARDES LOURENO
BARBOSA, CRISTIANI DE CASTILHO SANGUINETTE, ALDA LIMA
FALCO
Laboratrio de Leishmanioses - Centro de Pesquisas Ren 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 Cear [Cipa Group, available in
http://cipa.snv.jussieu.fr/; Aguiar & Medeiros, 2003. Distribuio e Hbitats. In:
Flebotomneos do Brasil (Rangel & Lainson Eds.), pp.207-257.]. This work aims to
discuss about the discovery of L. cruzi in Cear using specimens from the Municipality
of Quixad, deposited in the Collection of Phlebotomines of the Centro de Pesquisas
Ren 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 Cear 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 Cear, 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 MARN1,2, CRISTINA FERRO1,3, LEONARD MUNSTERMANN4,
CLARA OCAMPO1
1

Centro Internacional de Entrenamiento e Investigaciones Mdicas, CIDEIM., Cali,

Colombia
2
Laboratorio de Investigaciones en Parasitologa Tropica,l LIPT, Ibague, Colombia
3
Investigadora Emrita 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 Hidrologa, Meteorologa 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
CRISTINA FERRO1,2, HORACIO CADENA1, DAIRO MARN,1,3 , LEONARD
MUNSTERMANN4,CLARA OCAMPO1
1

Centro Internacional de Entrenamiento e Investigaciones Mdicas, CIDEIM., Cali,

Colombia
2
Investigadora Emrita Instituto Nacional de Salud, INS, Bogota, Colombia.
3
Laboratorio de Investigaciones en Parasitologa 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
GUTIRREZ MARN,1 NELSON FERNNDEZ,2 ALFONSO RODRIGUEZMORALES,3 CLAUDIA MAGALY SANDOVAL.1
1

Laboratorio de Entomologa Mdica, Instituto de Ciencias Biomdicas INBIOM,

Universidad de Pamplona, Colombia; 2Centro de Hidroinformtica, 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 Nio
(highest Oscillation Nio 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 Nio 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
Nio 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 JANABA,
MINAS GERAIS.
LARA-SILVA, F.O.1, MICHALSKY, E.M.1, 3, FRANA-SILVA, J.C.2, BARATA,
R. A.1, SOARES, L. F.1, DIAS, E. S.1
1 Centro de Pesquisas Ren Rachou / Fiocruz
2 Universidade Federal de Minas Gerais
3 Universidade Federal do Tringulo 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 Janaba.
Financial Support: FAPEMIG, UFTM, FIOCRUZ.

P-37
GEOGRAPHICAL DISTRIBUTION OF THE CORTELEZZI COMPLEX
(DIPTERA: PSYCHODIDAE: PHLEBOTOMINAE FROM BRAZIL).
GUSTAVO MAYR DE LIMA CARVALHO1, REGINALDO PEANHA
BRAZIL2, ALDA LIMA FALCO1, JOS DILERMANDO ANDRADE FILHO1.
1- Laboratrio de Leishmanioses - Centro de Pesquisas Ren Rachou - Fiocruz
2- Laboratrio de Bioqumica 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 Ren 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, Cear, and Maranho States), Southeast (Esprito Santo, Rio
de Janeiro, and So Paulo States) and South Regions (Paran 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, CDClike and Shannon trap, in peridomicile and in the forest, in Paran, Palmeirpolis, Peixe
and So 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 ORINOQUA - AMAZONA OF COLOMBIA
OLGA L. CABRERA1, LAUREANO MOSQUERA2, ERIKA SANTAMARA1,
CRISTINA FERRO1
1
2

Grupo de Entomologa, Instituto Nacional de Salud, Bogot, D.C., Colombia.

Secretara de Salud del Guaviare, San Jos 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
1

INSUE, National University of Tucumn (UNT), 2: Ministry of Health, 3CONICET,

4
REDILA: 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 reemergence 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, PER.
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 Per, 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 Convencin, Cusco, Per.
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 Maguia for their help during the study.

P-42
ABUNDANCIA RELATIVA DE ESPECIES DE LUTZOMYIA (DIPTERA:
PSYCHODIDAE) EN VEREDAS ENDMICAS PARA LTA, DEL NORESTE
COLOMBIANO
HOLVER PARADA 1, FAUSTINO MENDOZA1, CARLOS LVAREZ2,
REINALDO GUTIRREZ1, ROCIO CRDENAS1
1. Instituto de Investigaciones en Ciencias Biomdicas 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
transmisin activa de Leishmanisis Tegumenaria Americana LTA. Segn el Ministerio
de la Proteccin Social se ubic entre los cinco primeros departamentos con las mayores
tasas de transmisin para LTA 30.4/100.000 hab en el pas. Uno de sus municipios,
Abrego (bh-pM), con altura promedio de 1398 m.s.n.m y 21C, contiene veredas
endmicas para la transmisin de LTA y se mantiene en alto riesgo epidemiolgico
desde el ao 2003 segn estadisticas del IDS. En este estudio de 13 veredas con
pesquisas entomolgicas, 5 resultaron positivas para flebtomos: El Rosario, Los
Asientos, El Potrero, Mata de Fique y Anicillos. Consecutivamente durante 30 das, 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 mtodos de captura utilizados la trampa de
luz tipo Shannon fue la ms 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 asociacin indirecta de la poblacin de flebtomos a este tipo de
vegetacin. 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 asociacin con la ocurrencia de LTA en el municipio.

P-43
NEW
RECORDS
OF
LUTZOMYIA
FRANCA,
1924
PSYCHODIDAE) IN THE ORINOCO REGION OF COLOMBIA.

(DIPTERA:

RAFAEL J. VIVERO G1, LINA M. CARRILLO B1, ANDRS VLEZ1,

CAROLINA TORRES G1, SANDRA URIBE SOTO2, EDUAR E. BEJARANO3,
IVN D. VLEZ1
1. Research Associate, Program for the Study and Control of Tropical Diseases PECET, University of Antioquia. rajovigo2001@yahoo.com, Medelln, 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 Cao 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 JD2, PEANHA
BRAZIL R3
Instituto de Medicina Regional. Universidad Nacional del Nordeste, Chaco, Argentina,
e-mail: szelag_enrique@yahoo.com.ar
1
Ministry of Health/CONICET, Buenos Aires, Argentina.
2
Instituto Ren Rachou, Fiocruz, Belo Horizonte, MG, Brasil.
3
Instituto 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 Misin Nueva Pompeya
(2455S, 6130W) from November 2006 to December 2007 in Fortn 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 Referncia Nacional e Internacional para Flebotomneos of the Instituto Ren
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 EA1, SALOMON OD2, ANDRADE FILHO JD3,
CARVALHO GM3, STEIN M1, PEANHA BRAZIL R4
1

Instituto de Medicina Regional. Universidad Nacional del Nordeste. Chaco, Argentina.

e-mail: juan_rosa05@yahoo.com.ar
2
Ministry of Health/CONICET, Buenos Aires, Argentina.
3
Instituto Ren Rachou, Fiocruz, Belo Horizonte, MG, Brasil.
4
Instituto Oswaldo Cruz, Fiocruz, Rio de Janeiro, RJ, Brasil.
American Cutaneous Leishmaniasis (ACL) is endemic with occasional epidemics in
northern Argentinas 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 [Misin Nueva Pompeya (2455'S, 6130W)] and Wet
Chaco [Margarita Beln (2710`S,5850`W) and Resistencia (2726'S,5854'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 wasnt 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.
ROSA JR, SZELAG EA, STEIN M, SALOMON OD, SOBRADO SV1,
GORODNER JO
Instituto de Medicina Regional. Universidad Nacional del Nordeste. Resistencia,
Chaco, Argentina. juan_rosa05@yahoo.com.ar
2
Ministry 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 (2730S, 59W).
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(C)-rainfall(mm) was: Winter 49.82%15.7C-8.3mm due to an exceptional temperate dry (mean temperature 27C), Spring
21.52%-22.8C-10mm, Summer 15%-27C-14.2mm, Autumn 13.63%-19.9C-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 PAIVA2,
ROSELY
DOS
SANTOS
MALAFRONTE2,
MARIA
ELIZABETH
1
CAVALHEIROS DORVAL , REGINALDO PEANHA BRAZIL3, EUNICE
APARECIDA BIANCHI GALATI4
1. Universidade Federal de Mato Grosso do Sul, Laboratrio de
Parasitologia/DPA/CCBS, Campo Grande, MS, Brazil, E-mail: agoliveira@nin.ufms.br.
2. Universidade de So Paulo, IMT, So Paulo, Brazil. 3. Instituto Oswaldo Cruz- LBII,
Rio de Janeiro, RJ, Brazil. 4. Universidade de So Paulo, FSP, Departamento de
Epidemiologia, So 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 JANABA, STATE OF MINAS GERAIS,
BRAZIL
MICHALSKY E.M.1,3, FRANA-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
1

Centro de Pesquisas Ren Rachou/FIOCRUZ, 2 Universidade Federal de Minas Gerais,

3
Universidade Federal do Tringulo Mineiro, 4Secretaria Municipal de Sade de
Janaba, 5Fundao Ezequiel Dias; Minas Gerais, Brazil.
Visceral leishmaniasis (VL) has been reported in almost every state in Brazil. Until the
1970s, 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 Janaba, 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 Janaba.
Entomological captures were performed in 15 districts of Janaba 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
Janaba. 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 SANTAMARA1, ADOLFO VSQUEZ2, ANGLICA GONZLEZ2,
STELLA BUITRAGO3, AGUSTN GNGORA2, OLGA CABRERA1
1. Grupo de Entomologa, Instituto Nacional de Salud. Bogot 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 SOUZA2, ADRIANA ZWETSCH3;
CARLOS ROBERTO ALVES1, CONSTANA BRITTO1, ELIZABETH F.
RANGEL3
1

Lab.Biologia Molecular e Doenas Endmicas 3Lab. Transmissores de Leishmanioses,

Instituto Oswaldo Cruz, FIOCRUZ, Av.Brasil, 4365, Rio de Janeiro,RJ, Brasil 2Seo
de Reservatrios e Vetores, IPB, LACEN/RS, FEPPS, Secretaria Estadual de Sade 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 speciesspecific 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)
(2)

Universidad Nacional de San Antonio Abad del Cusco

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 COSTA1, CHERYL GOUVEIA1,
LUCIA H PINTO-DA-SILVA3, DEIVID C SOARES, ELVIRA M T SARAIVA2 &
E F RANGEL1
1
2

Laboratrio de Transmissores de Leishmanioses, Instituto Oswaldo Cruz/FIOCRUZ,

Departamento 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 (1x108 and 1x106 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, 1x105 and 1x106 CFU/mL, the viability of L. (L.) infantum chagasi was analyzed
in cytometer. The bacteria didnt demonstrate lysis effect on the promastigotes of both
Leishmania species, only the agglutinate effect was observed. However, in the coinfected 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 ATLNTICA (ATLANTIC FOREST)
(JACAREPAGU, RIO DE JANEIRO /RJ, BRAZIL).
CHERYL GOUVEIA1, ROSELY M. OLIVEIRA2 & ELIZABETH F. RANGEL1
1
2

Lab. Transmissores de Leishmanioses, IOC/FIOCRUZ

Dept. 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 Altntica, 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
mans 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.,CHVEZ O.,VIRLA J. & JUREZ E.
Postgrado en Educacin para la Promocin 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
IVONEIDE M. SILVA1, DAVI MARCOS S. DE OLIVEIRA1, IORLANDO R.
BARATA2, ADELSON A. A. SOUZA2, EDILENE OLIVEIRA DA SILVA1,
ELVIRA MARIA SARAIVA3
1

Universidade Federal do Par, 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 (Par), 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
BENITEZ,1,3 CARLOS FRANCO-PAREDES,4 ELINA ROJAS.1

JESUS

Instituto 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
Nio events occurred during 19942003 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 Nio events during years 1994, 1997 and 2002, and strong La Nia events
during 1995/1996 and 19982001. El Nio in eastern region of the country is expressed
as drought periods and La Nia 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 2281.1443.8 cases/year. During years with El Nio a mean
increase of 78.61% in ACL incidence was observed (ranging 8.3 to 614%), whilst in La
Nia 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 Nia) 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-ORTIGAO1, VINITA TRIPATHI1,
VALDIR BALBINO1, ILIANO COUTINHO-ABREU1, GWEN STAYBACK1,
MARIHA WADSWORTH1, EMIL LESHO5, EMAD DIN YEHIA FAWAZ2,
SHAABAN S. EL-HOSSARY2, HANAFI HANAFI2, DAVID HOEL2, MAHMOUD
ABO-SHEHDA3, SHADEN KAMHAWI4, GLENN WORTMANN5, AND MARY
ANN MCDOWELL1.
1

University of Notre Dame, Notre Dame, IN, USA

Naval Medical Research Unit #3, Cairo, Egypt
3
Jordan University of Science and Technology, Irbid, Jordan
4
LMVR, NIAID, NIH, Rockville, MD, USA
5
Walter Reed Army Medical Center, Washington DC, USA
2

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 nave 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 Parasitologa Experimental, Depto. Biologa, Facultad de
Ciencias Universidad de Los Andes, Mrida, EDO-Mrida, 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 g/pair of glands and for L.
ovallesi an average of 0.5 g/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 antimigonei 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
PHLEBOTOMINAE SAND FLIES (DIPTERA: PSYCHODIDAE)
DIFFERENT COLORED STICKY TRAPS.

OF
TO

BRAZIL RP1, BRAY DP2, TEMELJKOVITCH M3.

1

- Instituto Oswaldo Cruz,Fiocruz,Rio de Janeiro,RJ,Brazil

- Keele University, Staffordshire ,UK
3
- Secretaria de Agricultura, Prefeitura de Corumb, 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 Corumb, 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.
2

P-61
REEVALUATION OF THE FEEDING HABITS OF LUTZOMYIA
LONGIPALPIS IN THE CITY OF ARAATUBA, SO PAULO, BRAZIL,
FROM 2002 TO 2005, AFTER INTRODUCTION OF A DOG DELTAMETHRIN
COLLARS - SCALIBOR.
VERA LF CAMARGO-NEVES, MARLUCI GUIRADO, LLIAN AC RODAS,
GISELE S CABRAL, CLVIS PAULIQUVIS JR.
Leishmaniasis Study Group, Surveillance Epidemiologic Center, Endemic Control
Superintendence. Secretary of Health of State So Paulo, Brazil.
E-mail: veracamargo@saude.sp.gov.br
A previous study in Araatuba, So 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 -22C 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. SOUZA1, CHERYL
GOUVEIA1, VANDERLEI C. SILVA1, MARCELO S. GONZALES2 &
ELIZABETH F. RANGEL1
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 (AndradeCoelho 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 Sanit, Rome, Italy. E-mail:
marina.gramiccia@iss.it
Zoonotic Visceral Leishmaniasis (ZVL) incidence has been increased in Italy since
early 1990s 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, C2; MELO, C.M.1; ARAJO, E.D.1; CRUZ,
D.E.R.1; GES, M.A.O.1; PINTO, M.C.3
1

Instituto de Tecnologia e Pesquisa, Aracaju-SE; 2Superintendncia de Controle de

Endemias, Mogi Guau-SP; 3Faculdade de Cincias Farmacuticas, 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)
IVONEIDE M. SILVA1, IORLANDO R. BARATA2, ADELSON A. A. SOUZA2,
MARA C. PINTO3, CAROLINA SPIEGEL4, ANDREY J. ANDRADE5,
CLAUDIA C. GULIAS GOMES5, LVARO E. EIRAS5
1

Universidade Federal do Par, 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
(Par, 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 (10.943.71; 3.181.21) as well as UV light (7.531.78;
2.470.74). 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 Universitrio 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 3m. 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.
HERNNDEZ, MARA, DELGADO M, ALDANA A, MORN C, BRICEO 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

CAIZALES 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 Ncleo 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
AEZ 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
BRICEO 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
CAIZALES 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
DURSO V. OP-10
DVORAK VIT. OP-02

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

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

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

FAIMAN R. OP-26
FALCAO AL. OP-25, OP-32, P-05,
P-32, P-37
FAULDE MK. OP-28, OP-46

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
MAGUIA-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

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

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

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,

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

QUINTANA MG. OP-21, OP-22, P-40

QUISPE D. P-41
QUISPE M. P-52
QUISPE W. OP-54, P-41, P-52

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

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
YPEZ JY. OP-23

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

ZOLLNER GE. OP-52

ZWETSCH A. P-51

SPONSORS

Vicerrectorado de Investigacion (UPCH)