No description available.

No description available.

No description available.

No description available.

Les phlébovirus présentent sont présents dans toutes les régions du globe. Certains phlébovirus transmis par phlébotomes provoquent une maladie fébrile et des infections du système nerveux central. Depuis, de plus en plus de données montrent que la péninsule des Balkans joue un rôle majeur dans l'émergence de maladies à transmission vectorielle. Au début de ce travail, on comptait un nombre très limité de phlébovirus identifiés et isolés dans cette région. Une étude intégrée et transdisciplinaire en vue d'un inventaire des virus circulant dans pays des Balkans. (i) Un total de 3,850 phlébotomes sont été recueillis dans sept pays des Balkans en 2014 et 2015. Ils ont été testés pour la présence d'ARN viral et inoculé sur des cellules VERO afin d'isoler le virus détecté; (ii) des études de séroprévalence utilisant des tests de neutralisation ont été effectuées sur des échantillons de bovins et de moutons pour évaluer à deux agents pathogènes humains : le virus Toscana (TOSV) et le virus Sandfly fever Sicilian virus (SFSV). Nos résultats se composent de (i) la découverte et le séquençage de 3 nouveaux phlébovirus appartenant à 2 espèces différentes, (ii) la première identification du genotype B de TOSV en Croatie, (iii) la preuve de la co-circulation de deux genotypes (B et C) de TOSV, (iv) des taux d'anticorps neutralisants qui sont beaucoup plus élevés chez les bovins et les moutons pour le SFSV que pour TOSV. En conclusion, les résultats obtenus au cours de ce travail démontrent qu’es les Balkans représentent une zone de très importante activité pour les phlebovirus et donc mérite une surveillance particulière à cause du risque d’émergence et de dissémination. / Phleboviruses have a worldwide distribution. In the areas where sand flies are present, some of the sandfly-borne phleboviruses cause febrile illness and central nervous system infections. Sandfly fever was first reported in the Balkan Peninsula at the end of the 19th century. Since there is accumulating data showing that the Balkan peninsula plays a major role in the emergence of vector-borne diseases. At the outset of this work, a very limited number of phleboviruses had been identified and isolated in this region. To fill this gap, an integrated and transdisciplinary study was designed aiming at an inventory of viruses circulating in Balkans and associated seroprevalence studies using domestic animals: (i) a total of 3,850 sandflies were collected in seven Balkan countries (Albania, Bosnia-Herzegovina, Croatia, Kosovo, Montenegro, Republic of Macedonia and Serbia) in 2014 and 2015. They were tested for the presence of viral RNA and inoculated on VERO cell for virus isolation; (ii) seroprevalence studies using neutralisation tests were performed on cattle and sheep samples to assess the level of exposure to two human pathogens, Toscana virus (TOSV) and Sandfly fever Sicilian virus (SFSV). Our results consist of (i) the discovery and sequencing of 3 novel phleboviruses belonging to 2 different species, (ii) the identification for the first time of TOSV lineage B in Croatia, (iii) evidence of co-circulation of two lineages (Lineage B and C) of TOSV, (iv) rates of neutralising antibodies that are much higher in cattle and sheep for SFSV than for TOSV. Together the findings obtained during this work demonstrate that the Balkan area is a hot spot for phleboviruses.

Durant de nombreuses décennies, la faune phlébotomienne (Diptera, Psychodidae, Phlebotominae) Malgaches est demeurée très peu explorée. Deux Grassomyia avaient été signalés puis une espèce décrite sous le nom de Sergentomyia berentiensis. A partir des années 2000, la faune de Madagascar a révélé une richesse et une diversité non soupçonnées jusqu'alors avec la description de plusieurs espèces nouvelles et d'un sous-genre nouveau : Vattieromyia. Cette thèse est une contribution à la connaissance des Phlébotomes de Madagascar et des îles voisines des Seychelles et des Comores.Notre approche a été qualitative et non quantitative. Les phlébotomes collectés à Madagascar, aux Comores et aux Seychelles ont été étudiés morphologiquement puis, pour certains d'entre eux, par biologie moléculaire à diverses fins : associations mâles-femelles et systématique évolutive. Dans ce dernier cas, différents marqueurs ribosomiques, mitochondriaux et nucléaires ont été séquencés selon les problématiques.A Madagascar, les Phlebotomus forment un groupe monophylétique. Nous suggérons, sur des arguments morphologiques et moléculaires, de les individualiser dans un sous-genre nouveau étant donnée la mise en évidence de la paraphylie du sous-genre Anaphlebotomus dans lequel ont été classées les espèces malgaches.Nos travaux révèlent que P. fertei possède une aire de distribution qui couvre la presque totalité du pays. Les séquences de cytochrome b individualisent de nombreuses populations selon leurs origines géographiques mais nous n'avons pas pu individualiser ces populations sur le plan morphologique et morphométrique. Les séquences de l'ITS2 n'individualisent pas ces populations et nous critiquons l'utilisation du cytochrome b, et plus largement des marqueurs mitochondriaux, pour la systématique des Phlébotomes.En ce qui concerne les autres espèces de Phlebotomus, elles possèdent toutes une distribution étroite, réduite à leur lieu de capture. Nous avons décrit deux espèces nouvelles durant cette thèse : P. vaomalalae et P. vincenti. Les études moléculaires et morphologiques révèlent l'existence d'au moins trois espèces nouvelles : deux sympatriques à Andranoilovy (dont une espèce commune avec Berenty) et une à Ankililaoka.Enfin, nous proposons le rattachement de P. huberti au genre Sergentomyia. Cette espèce ne possède pas de soies mésanepisternales et le mâle que nous décrivons dans ce travail possède les caractères génitaux des Sergentomyia. De plus, nous décrivons sur une la seule femelle, une espèce nouvelle proche de S. huberti. Une étude moléculaire menée avec d'autres espèces supposées proches (appartenant au sous-genre Sintonius) nous conduit à proposer la création d'un nouveau sous-genre pour classer ces espèces malgaches.Nous analysons la paléobiogéographie des Phlébotomes de Madagascar et envisageons au moins deux épisodes de peuplement : l'un très ancien (environ 120 millions d'années), « africain » datant de la fragmentation du Gondwana et le second, plus récent (65 millions d'années), provenant d'Asie via un pont formé par le plateau des Seychelles.D'un point de vue épidémiologique, la recherche d'ADN leishmanien s'est révélée négative sur tous les phlébotomes testés.Dans l'archipel des Comores, aucun phlébotome n'avait été rapporté. Au cours de trois campagnes de piégeage menées en 2003, 2007 et 2011, nous rapportons la première mention de phlébotomes dans ces îles et décrivons deux taxons nouveaux S. pessoni et S. goodmani comorensis.Aux Seychelles, nous avons identifié S. clydei à Aldabra. Cette population possède des séquences mitochondriales très différentes des nombreuses populations continentales étudiées. L'origine du peuplement de cette île volcanique demeure mystérieuse, sans adéquation avec les données relatives à l'horloge moléculaire du cytochrome b dont nous doutons de la fiabilité. / During the last century, the Phlebotomine sand fly fauna (Diptera, Psychodidae, Phlebotominae) of Madagascar remained largely unexplored. Two Grassomyia were recorded and a species has been described as Sergentomyia berentiensis. From the 2000s, this fauna revealed a richness hitherto unsuspected: it included the description of several new species for Science and of a new subgenus (Vattieromyia). The present study is a contribution to the knowledge of Phlebotomine sand flies from Madagascar and the neighboring archipelagos of the Seychelles and the Comoros.The sand flies collected in Madagascar, the Comoros and the Seychelles were studied morphologically and, for some of them, by molecular biology in order to associate males with females and also to perform molecular systematics. Several molecular ribosomal, nuclear, and mitochondrial markers have been combined.In Madagascar, the Phlebotomus are grouped in a clade. Based on morphological characters and molecular studies, we suggest their individualization in a new subgenus because we show subgenus Anaphlebotomus where the Malagasy Phlebotomus were classified, is paraphyletic.P. fertei exhibits a wide distribution all over country. Sequences of cytochrome b individualize many populations linked to their geographical origins. However, it is not possible to individualize these populations based on morphological and morphometric characters. The sequences of ITS2 do not individualize these populations and we criticize the use of cytochrome b and other mitochondrial markers for the systematics of Phlebotomine sand flies.Regarding the other Malgaches Phlebotomus, all of them have a narrow distribution, reduced to their place of capture. We described two new species for Science: P. vaomalalae and P. vincenti. Moreover, molecular and morphological studies support the existence of at least three new species: two in sympatry in Andranoilovy (probably also recorded in Berenty) and one in Ankililaoka.Finally, we propose that P. huberti belongs to the genus Sergentomyia and not to the genus Phlebotomus. It does not have mesanepisternal setae and the male that we describe here exhibits Sergentomyia's genital characters. Moreover, we described on a female belonging to a new species close to S. huberti. We carried out a molecular study including continental species supposed closely related (belonging to the subgenus Sintonius). It individualizes the Malagasy specimens and consequently, considering their typical pharyngeal armature, we propose the creation of a new subgenus to classify them.We analyze the paleobiogeography of Malagasy sand flies. In agreement with generalized tracks, the settlement of Madagascar followed two routes at different times: one very old (about 120 million years ago), from "Africa" dating from the Gondwana fragmentation and the second, more recent (65 million years), from Asia using a bridge formed by the Seychelles plateau.From an epidemiological point of view, the search of Leishmania DNA was negative in all sandflies processed.In the Comoros Archipelago, no sand fly had been reported in the past. During three field works carried out in 2003, 2007 and 2011, we report the first record of sandflies in these islands and we describe two new taxa: S. pessoni and S. goodmani comorensis.In the Seychelles, we identified S. clydei in Aldabra. This population has mitochondrial sequences highly differing from those of many continental populations processed. The settlement of this volcanic island remains mysterious. They are not in agreement with molecular clock of cytochrome b sequences which seems of doubtful use.

Submitted by Ana Maria Fiscina Sampaio (fiscina@bahia.fiocruz.br) on 2013-04-18T18:08:55Z No. of bitstreams: 1 Ana Paula Almeida de Souza Validação de proteinas...pdf: 2348379 bytes, checksum: 16135e91d8da506bd6d51b00312ae853 (MD5) / Made available in DSpace on 2013-04-18T18:08:55Z (GMT). No. of bitstreams: 1 Ana Paula Almeida de Souza Validação de proteinas...pdf: 2348379 bytes, checksum: 16135e91d8da506bd6d51b00312ae853 (MD5) Previous issue date: 2013 / Universidade Federal da Bahia. Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz / O controle das populações de flebotomíneos, a contenção das epidemias nas fases iniciais e a facilitação do diagnóstico precoce dos indivíduos parasitados são estratégias traçadas pela OMS para a vigilância e controle da leishmaniose. Neste contexto, os testes sorológicos utilizando tanto o sonicado de glândula salivar (SGS) dos flebotomíneos quanto o antígeno solúvel de Leishmania (SLA) são importantes ferramentas para o controle/diagnóstico das Leishmanioses. No entanto, os testes realizados a partir da utilização de antígenos obtidos de frações brutas, apesar de apresentarem alta sensibilidade, alguns destes antígenos apresentam reação cruzada com epítopos compartilhados por outros patógenos/vetores. Além disso, o preparo dos antígenos não tem adequada reprodutibilidade devido a grande dificuldade de obtenção e/ou padronização destes. No presente trabalho, objetivamos identificar biomarcadores para a vigilância e o controle das Leishmanioses, utilizando proteínas recombinantes da saliva dos flebotomíneos como biomarcadores de exposição ao vetor e proteínas recombinantes da Leishmania para obtenção de um diagnóstico sorológico mais preciso para a Leishmaniose Tegumentar humana. Numa primeira etapa, foram realizados testes imunoenzimáticos contra as proteínas recombinante da saliva do vetor L. longipalpis (rLJM11 e rLJM17) para estimar a positividade anti-saliva num pequeno número de soros de crianças de uma área endêmica para Leishmaniose Visceral. Foi observado que os soros que reconhecem o SGS do L. longipalpis também reconhecem em diferentes proporções as proteínas rLJM17 e rLJM11. Ademais, as proteína recombinantes foram capazes de detectar a soroconversão anti-saliva de soros de uma segunda área endêmica para LV, havendo aumento no reconhecimento quando utilizadas as duas proteínas recombinantes de forma combinada. Além disso, a análise das curvas ROC evidenciou o desempenho superior da combinação das proteínas rLJM17 + rLJM11. Estes dados foram confirmados com a avaliação das proteínas frente um grande painel de 1.077 amostras de soro de indivíduos de uma outra área endêmica para LV. Nesta etapa, nossos resultados indicam que as proteínas rLJM11+rLJM17 representam uma ferramenta epidemiológica promissora que pode auxiliar na implementação de medidas de controle contra a Leishmaniose Visceral. Numa segunda etapa, testes imunoenzimáticos foram realizados contra uma série de proteínas recombinantes da Leishmania (HSP70, H2A, H2B, H3, H4 e KMP11) a fim de selecionarmos proteínas antigênicas contra soros de pacientes com Leishmaniose Cutânea (LC) e Leishmaniose Mucosa (LM) e que apresentassem elevada especificidade quando testadas contra soros de indivíduos com outras patologias (doença de Chagas, Lúpus Eritematoso Sistêmico, Hanseníase e Tuberculose). Para avaliar a eficácia das proteínas recombinantes foram utilizadas curvas ROC, possibilitando a seleção de antígenos possivelmente mais eficientes que o SLA no imunodiagnóstico da Leishmaniose. As proteínas recombinantes HSP70 e H2A foram selecionadas por apresentarem elevada sensibilidade, sendo reconhecida por anticorpos dos soros de pacientes com LM e LC respectivamente. Na última etapa dos experimentos, utilizando soros de indivíduos que apresentavam outras patologias, observou-se que a reação cruzada diminui frente aos antígenos recombinantes, especialmente para a rHSP70, quando comparamos à observada para o SLA. Nossos resultados mostram a elevada antigenicidade da rHSP70, sugerindo a possibilidade de utilização desta proteína recombinante para o sorodiagnóstico da Leishmaniose Tegumentar. Neste trabalho foi possível identificar e validar o uso de proteínas recombinantes do parasito e da saliva dos flebotomíneos como biomarcadores para o sorodiagnóstico e avaliação da exposição às leishmanioses. / The population control of sand flies, containment of epidemics in the early stages and facilitating early diagnosis of infected individuals are strategies outlined by WHO for surveillance and control of Leishmaniasis. In this context, serologic tests using both the salivary gland sonicate (SGS) of sandflies as soluble Leishmania antigen (SLA) are important tools for the control / diagnosis of Leishmaniasis. However, tests based on the use of antigens derived from crude fractions, despite showing high sensitivity, some of these antigens cross-react with epitopes shared by other pathogens / vectors. Moreover, the preparation of antigens has adequate reproducibility due to difficulty in obtaining and / or standardization of these. In this study, we aimed to identify biomarkers for the surveillance and control of Leishmaniasis, through use of recombinant proteins from the saliva of sandflies as biomarkers of exposure to vector and through the use of recombinant proteins of Leishmania to get a more accurate serodiagnosis for human leishmaniasis. On the first step, ELISAs were performed against the recombinant protein from the saliva of the vector L. longipalpis (rLJM11 and rLJM17) to estimate the positive anti-saliva on a small number of sera from children from an endemic area for VL. It was observed that the sera that recognize the SGS of L. longipalpis also recognize in different proportions of rLJM17 and rLJM11 proteins. Further, each recombinant protein was able to detect anti-saliva seroconversion in sera from a second endemic area for VL, with increase in recognizing when the two recombinant proteins were used in combination. Furthermore, the ROC analysis showed the superior performance of the combination of rLJM17 + rLJM11, these data confirmed using a large panel of 1077 serum samples from individuals from another endemic area for LV. At this stage, our results indicate that proteins rLJM11 + rLJM17 represent a promising epidemiological tool that can assist in the implementation of control measures against Visceral Leishmaniasis. In a second step, ELISAs were performed against a series of recombinant proteins of Leishmania (HSP70, H2A, H2B, H3, and H4 KMP11) so antigenic proteins were selected against serum from patients with cutaneous leishmaniasis (CL) and mucocutaneous leishmaniasis (MCL) and that presented high specificity when tested against sera from patients with other diseases (Chagas disease, Systemic Lupus Erythematosus, Leprosy and Tuberculosis). To identify the effectiveness of recombinant proteins, ROC curves were used to select antigens possibly more efficient than SLA. Recombinant proteins HSP70 and H2A were selected for having high sensitivity in recognizing sera from patients with LM and LC respectively. In the last experimental stage using sera from individuals with other pathologies, it was observed that the reduced cross-reactivity against the recombinant antigen, especially for rHSP70 when compared to that observed for the SLA. Our results show the high antigenicity of rHSP70, suggesting the possibility of using this recombinant protein for serodiagnosis of Leishmaniasis. In this work, it was possible to identify and validate the use of recombinant proteins of the parasite and of the saliva of sandflies as biomarkers for serodiagnosis and assessment of exposure to Leishmaniasis.

Made available in DSpace on 2015-04-20T12:31:31Z (GMT). No. of bitstreams: 1 Francimeire Gomes.pdf: 3198532 bytes, checksum: 51d8088daaeb9e42ea2339b8f0578ac7 (MD5) Previous issue date: 2013-06-07 / Fundação de Amparo à Pesquisa do Estado do Amazonas / Apesar do conhecimento produzido sobre os diversos elementos que compõem a cadeia de transmissão da Leishmaniose Tegumentar Americana (LTA) no Amazonas, os estudos relacionados a tríade epidemiológica (hospedeiro, vetor e agente etiológico) desta doença nos municípios da região conhecida como Alto Rio Negro (São Gabriel da Cachoeira - Santa Izabel do Rio Negro Barcelos) ainda são escassos, em particular em São Gabriel da Cachoeira (SGC), onde a região limita-se com os paises da Venezuela e Colômbia. Em decorrência disso e considerando a ausência de trabalhos relacionados ao agente etiológico e vetores da LTA que se encontram circulando no município de SGC, foi realizado levantamento da entomofauna de flebotomíneos e com isto verificou-se os potenciais transmissores de tripanossomatídeos e/ou leishmânias; além disso, avaliou-se a infecção natural nestes insetos pela técnica de dissecção e Nested-PCR, e finalmente realizou-se a caracterização bioquímica e molecular para identificação das espécies de Leishmania, isoladas de flebotomíneos e humanos. Do total de 6832 flebotomíneos coletados e distribuídos em 50 espécies foi observado um índice de diversidade considerável (H = 1,19). Destes 66% (4526/6832) correspondem a 19 espécies envolvidas na transmissão de leishmânias e/ou tripanossomatídeos de importância para a saúde pública. Foi notificado o primeiro registro de Lutzomyia conviti para o Brasil e para o Amazonas. A taxa de infecção natural por tripanossomatídeos foi em torno de 4,26%, sendo L. dendrophyla a espécie que apresentou a maior taxa de infecção, com cerca de 3,6%. Dos isolados, apenas 24 cresceram em cultivo, sendo 23 de flebotomíneo e um humano. Um total de 470 fêmeas de 13 espécies diferentes entre elas: L. ayrozai, L. dendrophyla e L. davisi foram testadas com os iniciadores descrito por Cruz et al. (2002). Os isolados não apresentaram perfis bioquímicos similares as cepas de referência de Leishmania e Endotrypanum. Dos fragmentos de DNA da região ITS sequenciados, verificou-se a formação de três grupos/espécies, sendo nove com características biológicas similares ao subgênero Viannia e dois para o subgênero Leishmania. Sugere-se a participação de L. dendrophyla na transmissão de flagelados do gênero Leishmania circulando em SGC, podendo ser a região considerada como área de risco em LTA.

Circulation des Phlébovirus en Turquie, l'Iran et l'Algérie a été étudiée. L'isolement, la caractérisation génomique, les relations phylogénétiques de six virus ont été présentées: le virus Adana (ADAV), deux souches de virus Toros (TORV), le virus Zerdali (ZERV) de la Turquie; le virus Dashli (DASHV) de l'Iran; le virus Toscana (TOSV) d'Algérie. Cette étude a commencé avec la collection de 38,131 phlébotomes de la nature. La méthode de séquençage de nouvelle génération (NGS) à haut débit nous à été utilisée pour l’analyse des génomes complet des virus isoles. En conclusion, cette étude a d'importantes contributions sur phlébovirus négligées. Voici quelques-unes des contributions significatives; (i) ZERV et TORV qui sont étroitement apparentés au virus Tehran (THEV) et le virus Corfou (CFUV), respectivement, ont été isolés depuis 56 et 30 ans des premiers isolements de THEV et CFUV, respectivement, (ii) Détection du virus ADAV un animal domestique et sur quelques sérums humain par test de neutralisation. Ce virus ADAV constitue avec le virus le virus (SALV), le virus Arbia (ARBV), et le virus (ADRV) le groupe Salehabad. Seul le virus ADRV a été détectée dans le liquide cérébro-spinal auparavant landais que avec les autres, aucune preuve pathogène n’a été détectée, (iii) Nous avons découvert la plus récente circulation phlébovirus en Iran après 56 années, (iv) TOSV a été isolé en Algérie pour la première fois et la circulation a été confirmée par séropositivités dans le sérum humain. / Sandfly-borne phlebovirus circulation in Turkey, Iran, and Algeria was investigated. The isolation, genomic characterization, phylogenetic relationships of 6 viruses was presented: Adana virus (ADAV), two strains of Toros virus (TORV), Zerdali virus (ZERV) from Turkey; Dashli virus (DASHV) from Iran; Toscana virus (TOSV) from Algeria. This study has begun with the collection of 38,131 sandflies from nature. The well established, high-throughput methodology was applied for the discovery of viruses including PCR tools and cell culture methods. Next generation sequencing (NGS) technology facilitated to perform complete genome analysis of the isolated viruses. In conclusion, this study has contributions to the neglected sandfly-borne phlebovirus group and filled some gaps about the circulation of these agents in Turkey, Iran, and Algeria. Following are some significant contributions; (i) ZERV and TORV which are closely related to Tehran virus (THEV) and Corfou virus (CFUV), respectively were isolated after 56 and 30 years of the first isolations of THEV and CFUV, respectively, (ii) There was no evidence of the pathogenicity of Salehabad virus (SALV) and Arbia virus (ARBV) except the detection of Adria virus (ADRV) in CSF until ADAV which belongs to the Salehabad serocomplex was detected in domestic animal and very few human sera by neutralization assay, (iii) We have discovered the most recent sandfly-borne phlebovirus circulation in Iran after 56 years, (iv) TOSV was isolated in Algeria for the first time and circulation was confirmed by seropositivities in human sera.

Made available in DSpace on 2015-03-26T13:30:27Z (GMT). No. of bitstreams: 1 texto completo.pdf: 6083054 bytes, checksum: 0c926b8830bf109aea4cf3a77c1bcbba (MD5) Previous issue date: 2009-04-28 / Conselho Nacional de Desenvolvimento Científico e Tecnológico / The sandfly are diseases caused by protozoa, intracellular binding, plagued heteroxenic, pathogenic to humans, belonging to the genus Leishmania Ross,.These are considered serious human protozoan public health problems worldwide, affecting indiscriminately men, women and children, causing different clinical manifestations. Its distribution reaches the tropical and subtropical regions of America, Africa, Asia, Europe. According to the World Health Organization (WHO), we observed the occurrence of this disease in 88 countries, of which 16 are developed. Currently it is estimated there, worldwide, around 350 million people in areas at risk of both leishmaniasis (WHO, 2006). In the Americas, the visceral and cutaneous leishmaniasis, have wide distribution, with records of cases from the extreme south of the United States to northern Argentina, with the exception of Chile and Uruguay. The visceral leishmaniasis occurs from Mexico to Argentina, however, was not registered yet, Uruguay, in the Andean countries, Chile, Peru, Ecuador, and northern South America, French Guiana, Guyana and Suriname (LAISON & Shaw, 1998). In the Brazilian territory, the leishmaniasis are included in the list of diseases that make up the Schengen Information System Diseases of Compulsory Notification (SNDC), and Brazil considered the country with the highest prevalence of these. / Os flebotomíneos, vetores das leishmanioses, são identificados e classificados pela morfologia dos insetos adultos, existindo poucos conhecimentos de caracteres das formas imaturas. Das 500 espécies Neotropicais descritas, somente 66 espécies tem suas formas imaturas estudadas. Visando avaliar a importância de alguns caracteres taxonômicos dos estádios imaturos de flebotomíneos, estudos morfológicos foram realizados em Lutzomyia (Nyssomia) intermedia e L. (N.) neivai, transmissores de Leishmania (V.) braziliensis, de populações procedentes dos Estados do Rio de Janeiro e São Paulo, que durante alguns anos estiveram em sinonímia e têm apontado certo grau de introgressão. Em L. (N.) neivai pode-se observar um maior número de tubérculos formando uma ou mais fileiras, quando comparados aos observados no exocórion de L. (N.) intermedia. Em características gerais, as larvas, das duas espécies, apresentaram-se bastante semelhantes. Porem, as larvas do primeiro estádio revelaram diferenças significativas com relação aos outros estádios evolutivos, destacando a presença de egg buster, o tamanho das antenas e o número de cerdas caudais. O uso destes caracteres não se traduziu em uma boa ferramenta para separação das espécies objeto do nosso estudo.