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Canine visceral leishmaniasis (CVL) is the major source of human visceral leishmaniasis (VL) and is transmitted from dogs to sand flies to humans. To control the spread of this disease, early and accurate detection of infected dogs is critical but challenging. Here we demonstrate the potential of the Dual-Path Platform (DPP(®)) CVL rapid test for detecting K26/K39-reactive antibodies in sera from clinically symptomatic (n=60) and asymptomatic (n=60) Leishmania infantum-infected dogs. For the specificity evaluation, assays were performed using known negative diagnostic serum samples (n=59) and cross-reaction control sera (n=11) from animals born in a VL-free area of Brazil. The diagnostic kit displayed high specificity (96%) but low sensitivity (47%) in identifying parasite-positive dogs without signs of CVL. However, the test sensitivity was significantly higher (98%) in diseased cases, indicating that this convenient test may be useful to identify the most infectious dogs. Efforts should be pursued to obtain a more sensitive DPP-multiplexed test parameter (i.e. based on simultaneous yet separate antibody detection of carefully selected multiple antigens of diagnostic utility) for effective serodiagnosis of early-infected dogs, as this will likely allow more efficient canine removal regimens than those used in practice by public health services.

A new species of Sycorax Curtis, 1839 (Diptera, Psychodidae) from the Atlantic Forest in southeastern Brazil. Sycorax bravoi Santos, Ferreira & Falqueto sp. nov. is described and illustrated based on samples collected with a Möricke trap installed on the ground at the Biological Station of Santa Lúcia, municipality of Santa Teresa, in the Brazilian state of Espírito Santo. Males have a paramere with a spiniform prolongation on the distal surface and an aedeagus with a long posterior membranous dorsal prolongation. Females have a racket-shaped genital furca and tubular spermatheca, tapered on the apical third. This finding raises the number of Sycorax species known from Brazil to seven. Espécie nova de Sycorax Curtis, 1839 (Diptera, Psychodidae) da Mata Atlântica, sudeste do Brasil. Sycorax bravoi Santos, Ferreira & Falqueto sp. nov. é descrita e ilustrada a partir de exemplares coletados com armadilha Möricke instalada sobre o solo, na Estação Biológica de Santa Lúcia, município de Santa Teresa, estado do Espírito Santo, Brasil. Machos apresentam parâmero com prolongamento espiniforme na superfície distal e edeago com longo prolongamento posterior membranoso e dorsal. Fêmeas com forquilha genital em forma de raquete e espermateca tubular, afilada no terço apical. Eleva-se para sete o número de espécies de Sycorax para o Brasil.

INTRODUCTION: This study aimed to evaluate the presence of Aedes aegypti in breeding sites located in vacant lots (VLs) and determine the effectiveness of VL cleaning to reduce insect foci. METHODS: Two types of VLs were sampled, the experimental VL, which was cleaned monthly, and the control VL, which was not cleaned. RESULTS: Monthly cleaning of VLs reduced the abundance of immature forms of A. aegypti. CONCLUSIONS: Strategies for combating this vector should include regular cleaning of VLs and educating the public regarding the risks of discarding waste in inappropriate areas. INTRODUÇÃO: Os objetivos deste estudo foram avaliar a ocorrência de Aedes aegypti em diferentes tipos de criadouros em terrenos baldios (TBs) e os efeitos da limpeza mensal do terreno na redução dos focos do inseto. MÉTODOS: Dois tipos de TBs foram amostrados, TB experimental submetido à limpeza mensal e TB controle, sem limpeza prévia. RESULTADOS: A limpeza mensal dos TBs reduziu significativamente a abundância de formas imaturas do inseto. CONCLUSÕES: Estratégias de combate ao vetor devem incluir a limpeza de TBs e a conscientização da população sobre o risco que representa o descarte de lixo em local impróprio.

This study evaluates the importance of native bromeliads growing on rocky outcrops interspersed with urbanized areas as breeding sites for the Aedes aegypti (L.) in Vitória, state of Espírito Santo, Brazil. Oviposition traps were installed in backyards of houses in two separate zones. In the first zone houses were up to 50 m away from the rocky outcrops, while in the second zone they were at least at 200 m from the rocky outcrops. Aedes aegypti was significantly more abundant in the latter zone. The finding was that rocky outcrops with native bromeliads, even with the greater availability of potential breeding sites, do not play an important role as breeding sites for A. aegypti. This conclusion supports the hypothesis that the macrobiota of native bromeliads plays an important role in the natural control of A. aegypti. Besides, the interspecific competition between species of mosquitoes and the attractiveness of bromeliads could also be important factors.

Aim The aim of this study was to test the hypothesis that the dispersal of sylvatic triatomines to anthropic habitats is stimulated by degradation of their natural habitat. Location State of Espirito Santo, south-eastern Brazil. Methods We georeferenced records of domiciliary captures of sylvatic triatomines (1996–2005) obtained from the database of the entomological surveillance programme of the Brazilian National Health Foundation. We partitioned the study area into 543 equal hexagons and attributed information on the presence/absence of triatomines and climatic, topographic and biological variables to each. Using generalized additive models (GAMs), we defined the determinant variables for triatomine presence/absence and demarcated the potential areas in which they might occur. We then subjected the hexagons with a high probability of triatomine presence to a second analysis, adding information on the number of domiciliary captures of triatomines and landscape indices that measured quantitative and spatial patterns of forest fragments. We used GAMs to determine the indices that influenced the number of domiciliary captures of triatomines. Results The presence of triatomines was characterized by a strong positive correlation with variability of terrain slope. This variable was used to demarcate the areas where triatomines were likely to occur. Landscape indices associated with the number of domiciliary captures of triatomines were the number of patches, the fractal dimension index, the total core area and the clumpiness index. Main conclusions Our results do not support the hypothesis that domiciliary invasion by sylvatic triatomines is stimulated by environmental degradation. Instead, the results suggest that less degraded areas maintain larger populations of triatomines and consequently present higher indices of dispersal and domiciliary invasion. Particular landscape patterns and dispositions of human dwellings, however, may enhance the probability of triatomines being attracted to the artificial light of dwellings. Initiation of flight and consequent dispersal is a naturally occurring process that allows triatomines to find and colonize areas that favour the survival of their offspring.

Our objective was to study and evaluate the richness and diversity of Phlebotominae fauna in the Duas Bocas Biological Reserve (DBBR) in the state of Espírito Santo, in southeastern Brazil. Sand fly collections were carried out during four consecutive nights each month between August 2007 and July 2008 at DBBR by using CDC automatic light traps and an illuminated Shannon trap. Specific richness (S) and Shannon diversity index (H) was calculated for each trap. We collected 18,868 sand flies belonging to 29 species and 13 genera. Nyssomyia yuilli yuilli was the most abundant species followed by Psychodopygus ayrozai, Ps. hirsutus, Psathyromyia pascalei, and Ps. matosi. We recorded Brumptomyia cardosoi, Br. troglodytes, and Ps. geniculatus for the first time in the state of Espírito Santo. We discuss the differences in diversity and richness of the sand flies in both traps and in relation to other Brazilian localities and biomes. We also discuss the possibility of wild transmission of Leishmania in the DBBR and the influence of the sand fly species in leishmaniasis transmission to the adjacent areas of the reserve.

This paper describes sand flies similar to Nyssomyia intermedia (Lutz & Neiva) with variations in the number of spines at the gonostyle and tests the hypothesis whether these specimens belong or not to N. intermedia species. Using Principal Component Analysis and Neighbour Joining, the measurements of 15 structures of the phlebotomine with variations in number of spines were compared with measurements of 30 sand flies of N. intermedia species. Both analyses didn't cluster the specimens with variation in spines number in one group and provided evidence to the hypothesis that those sand flies belong to N. intermedia.

Entre os anos de 2004 e 2008 foram realizadas coletas de Simuliidae em várias localidades no Estado do Espírito Santo, Brasil. No total, 66 córregos e rios das doze principais bacias hidrográficas do estado foram amostrados. Dezessete espécies foram coletadas, sendo que nove delas representam ocorrências novas para o estado (Simulium pertinax, S. jujuyense, S. rubrithorax, S. subnigrum, S. travassosi, S. spinibranchium, S. hirtipupa, S. lutzianum e S. anamariae), duplicando o número de espécies conhecidas no estado. We conducted a freshwater survey from 2004 to 2008 in several localities of Espírito Santo, Brazil. In total, 66 streams and rivers were sampled in the 12 hydrographic basins of the state. We collected and identified 17 species of which nine represent new records (Simulium pertinax, S. jujuyense, S. rubrithorax, S. subnigrum, S. travassosi, S. spinibranchium, S. hirtipupa, S. lutzianum e S. anamariae). These findings doubled the number of records of black fly species to the State of Espírito Santo, Brazil.

Free-living amoeba of the genus Acanthamoeba can eventually act as parasites, causing infections in humans. Some physiological characteristics of Acanthamoeba have been related to the grade of pathogenicity, allowing inferences about the pathogenic potential. The main goal of this study was to characterize isolates of Acanthamoeba obtained in Brazil and evaluate properties associated with their pathogenicity. A total of 39 isolates obtained from keratitis cases (n = 16) and environmental sources (n = 23) were classified into morphological groups and genotyped by sequencing the 18S rDNA fragments ASA.S1 and GTSA.B1. Samples were also tested regarding their thermo-tolerance, osmo-tolerance, and cytopathogenicity in MDCK cells. Isolates were identified and classified as follows: group I (T17, T18); group II (T1, T3, T4, T11); and group III (T5, T15), with the predominance of genotype T4 (22/39). Clinical isolates were genotyped as T3 (1/16), T4 (14/16) and T5 (1/16). The majority of isolates (38/39) were able to grow at 37 °C, but tolerance to 40 °C was more frequent among environmental samples. The tolerance to 1 M mannitol was infrequent (4/39), with three of these corresponding to clinical samples. The variable ability to cause cytopathic effects was observed among isolates of distinct genotypes and origins. This study identified, for the first time, T1, T15, and T18 in Brazil. It also indicated a weak association between the clinical origin of the isolates and tolerance to high temperatures, high osmolarity, and cytopathogenicity, demonstrating that some in vitro parameters do not necessarily reflect a higher propensity of Acanthamoeba to cause a disease.

Lutzomyia Franca is the largest and most diverse sand fly genus in the New World and contains all the species involved in the transmission of American visceral leishmaniasis (AVL). Morphological characters were used to test the monophyly and to infer phylogenetic relationships among members of the Lutzomyia subgenus. Fifty-two morphological characters from male and female adult specimens belonging to 18 species of Lu. (Lutzomyia) were scored and analyzed. The resulting phylogeny confirms the monophyly of this subgenus and reveals four main internal clades. These four clades, however, do not support the classification of the subgenus in two series, longipalpis and cavernicola, because neither is necessarily monophyletic. Knowledge on phylogenetic relationships among these relevant vectors of AVL should be used as a tool for monitoring target taxa and a first step for establishing an early warning system for disease control.

Santos, Claudiney Biral Dos, Falqueto, Aloísio, Alexander, Bruce (2009): Description of three new species of Nemapalpus (Diptera, Psychodidae, Bruchomyiinae) from Atlantic Forest in southeastern Brazil. Zootaxa 2264: 58-64, DOI: 10.5281/zenodo.190795

In an endemic rural area of southeast Brazil, surveys confirmed that dogs serve as peridomestic reservoirs of Leishmania infantum. It is likely that the lack of efficient control is because presently used diagnostic tests miss positive dogs. Overall, 57% of the dogs had specific antibodies, but the canine infection was not uniformly fatal and many seropositive dogs remained asymptomatic or even spontaneously recovered. Furthermore, 42% of the human residents became leishmanin-positive reactors and 47% had positive serology at the initial survey, but our estimates also point at a high recovery rate among the infected population with time. The delayed-type hypersensitivity (DTH) reaction to Leishmania was a good indicator of resistance to infection in this particular epidemiologic situation. The lack of any significant differences in infection rates by gender or age indicate that all of the population was at an equal risk of infection and most people were infected in the peridomestic setting.

Panstrongylus geniculatus is one of the most widely distributed species of Triatominae in South and Central America, occurring in several biomes with different climatic characteristics. We present the results of captures of P. geniculatus provided from a 10-year program of entomological surveillance in the state of Espírito Santo, Brazil. We also present an updated geographic distribution map of P. geniculatus obtained from an extensive literature review. The entomological surveillance program, between 1996 and 2005, provided 110 capture registers of P. geniculatus in Espírito Santo. These are the first records of P. geniculatus from 15 municipalities in this state. P. geniculatus extends through 18 countries in Central and South America, from southern Mexico to northern Argentina.

The diagnosis of visceral leishmaniasis remains difficult in rural areas where the disease is endemic, and serologic methods still need assessment, as they are not very sensitive for the detection of asymptomatic infectious dogs. Here we present data on the development of enzyme-linked immunosorbent assay (ELISA)-based methods for the detection of antibodies against recombinant leishmanial antigens (namely, the recombinant K26 [rK26] and rK39 antigens from Leishmania infantum and the rA2 protein from Leishmania donovani ) in comparison to ELISAs employing crude soluble antigen (CSA). The assays utilized sera from known negative controls ( n = 25) and clinically asymptomatic ( n = 50) and symptomatic ( n = 50) dogs with confirmed L. infantum infections. Additional studies were also done using sera from animals harboring other infections ( n = 14) for the evaluation of cross-reactivity. Our study indicated that rK26 and rK39 used in ELISAs provided very high sensitivities for the detection of symptomatic dogs (94% and 100%, respectively), followed by CSA (88%) and rA2 (70%). Conversely, rA2 was more sensitive for asymptomatic dogs (88%) than rK39 and rK26 (both 66%) and CSA (30%). Some cross-reactivity in sera from dogs with other infections ( Leishmania braziliensis and Leptospira interrogans ) was identified, but the rA2 protein provided the greatest specificity (98%). Data further indicate that all three recombinant proteins must be used in parallel to detect essentially all infected dogs. Efforts should be made to develop a cheap and reliable serologic test based on epitope selection from these diagnostic markers for the sensitive detection of L. infantum -infected dogs.

Randomly amplified polymorphic DNA (RAPD) analysis was performed to infer the magnitude and pattern of genetic differentiation among Lutzomyia (Nyssomyia) intermedia populations from two ecologic regions in southeast Brazil where transmission of American cutaneous leishmaniasis (ACL) by L. (Viannia) braziliensis reflects distinct eco-epidemiologic features. Two hundred thirty-five bands from six primers were analyzed for genetic variation within and between sand fly populations. A lower level of intrapopulational variability was found in domestic sand flies (compared with the peridomestic population). Gene flow FST estimates indicated homogeneity in the studied population, with limited population substructuring, but with a tendency of sand fly vectors to form clusters by micro-habitat (domestic and peridomestic). The level of genetic structuring of Lu. intermedia population from each ecologic region is distinct and may reflect association or independence between the domestic and the peridomestic habitats in rural and periurban areas, respectively, both reflecting distinct characteristics of the transmission cycle of ACL.

Adults of Triatoma vitticeps infected by flagellates similar to Trypanosoma cruzi are frequently captured by the inhabitants of rural areas in the Brazilian state of Espirito Santo. The dynamics of feeding and defecation were observed in three groups of adult triatomines, consisting of sylvatic T. vitticeps and laboratory-reared specimens of this species and T. infestans. Triatomines were observed from the moment they were presented with an immobilized chicken as a bloodmeal source until 240 min after feeding had ended. Mean times between the end of feeding and defecation for T. infestans, wild T. vitticeps and laboratory-reared specimens of the latter species were 1.2, 21.1, and 64 min respectively. All T. infestans defecated within 10 min of feeding, while only 29.9 of wild and 52.8% laboratory-reared specimens of T. vitticeps did so within this period. These results may explain the low efficiency of T. vitticeps in T. cruzi transmission to man. The shorter time between feeding and defecation in laboratory-reared T. vitticeps may indicate a change in behaviour of this species as a result of adaptation to an artificial environment.

In Brazil, the main etiologic agent of Leishmaniasis that frequently presents with mucosal involvement belongs to the Viannia subgenus. The therapeutic conduct in this disease depends on the parasitological diagnosis, and classical methods are restricted in identifying the agent. In this paper we describe a polymerase chain reaction (PCR), which uses primers designed from mini-exons repetitive sequences. The PCR amplifies a 177bp fragment that can distinguish (Viannia) from (Leishmania) subgenus. This test could be a useful diagnostic tool.

Paracoccidioidomicose é a micose sistêmica endêmica mais frequente no Brasil. No início, o paciente não desenvolve sintomas. Com a progressão da doença, o indivíduo pode apresentar envolvimento disseminado, sendo que o acometimento ósseo é extremamente raro. O objetivo deste artigo é avaliar as alterações ósseas encontradas em estudos de imagem em um paciente com osteomielite de punho decorrente de paracoccidioidomicose disseminada.

Background: Because domestic dogs are reservoir hosts for visceral leishmaniasis (VL) in Brazil, one of the approaches used to reduce human disease incidence is to cull infected dogs. However, the results of controlled intervention trials based on serological screening of dogs and killing of seropositive animals are equivocal. A prophylactic vaccine to protect dogs from being infectious to the sand fly vector could be an effective strategy to provide sustained control. Here, we investigated whether a currently licensed commercial subunit rA2 protein-saponin vaccine (Leish-tec®) had an additional effect to dog culling on reducing the canine infectious populations. Methodology/principal findings: This prospective study was conducted in an L. infantum highly endemic area of southeast Brazil. At the onset of the intervention, all of the eligible dogs received through subcutaneous route a three-dose vaccine course at 21-day intervals and a booster on month 12. For the purpose of comparison, newly recruited healthy dogs were included as the exposed control group. To ascertain vaccine-induced protection, dogs were screened on clinical and serological criteria every 6 months for a 2-year follow-up period. Antibody-based tests and histopathological examination of post-mortem tissue specimens from euthanized animals were used as a marker of infection. The standardized vaccine regime, apart from being safe, was immunogenic as immunized animals responded with a pronounced production of anti-A2-specific IgG antibodies. It should be noted the mean seroconversion time for infection obtained among immunized exposed dogs (~ 18 months), which was twice as high as that for unvaccinated ones (~ 9 months). After two transmission cycles completed, the cumulative incidence of infection did differ significantly (P = 0.016) between the vaccinated (27%) and unvaccinated (42%) dogs. However, the expected efficacy for the vaccine in inducing clinical protection was not evident since 43% of vaccine recipients developed disease over time. Our estimates also indicated that immunoprophylaxis by Leish-tec® vaccine in addition to dog culling might not have an impact on bringing down the incidence of canine infection with L. infantum in areas of high transmission rates. Conclusions/significance: Leish-tec® as a prophylactic vaccine showed promise but needs to be further optimized to be effective in dogs under field conditions, and thereby positively impacts human incidence.

We evaluated the ecological and epidemiological aspects of the sand fly fauna in an area of the Atlantic Forest biome with records of visceral and cutaneous leishmaniasis. Sand fly collections at three different localities at the National Monument of Pontoes Capixabas, State of Espirito Santo, Southeastern Brazil, were conducted by using two Centers of Disease Control and Prevention automatic light traps in the peridomiciliary environment and eight Centers of Disease Control and Prevention automatic light traps in the forested environment. Collections occurred during four consecutive nights within each of the months and locations: Sao Luiz (December 2009, May 2010, July 2010, and December 2010), Corrego Palmital de Baixo (September 2010 and October 2010), and Corrego Sao Bento (February 2011 and May 2011). We collected 21,138 sand flies belonging to 31 species and 14 genera. Of this total, 12,412 sand flies were captured in the peridomiciliary environment and 8,726 in the forested environment. All of the vector species, Lutzomyia longipalpis (=Lutzomyia longipalpis, sensu; Young and Duncan), Migonemyia migonei (=Lutzomyia migonei, sensu; Young and Duncan), and Nyssomyia intermedia (=Lutzomyia intermedia, sensu; Young and Duncan), occurred in significantly higher numbers in the peridomiciliary environment than compared with the forested environment. Our results highlight the importance of conservation in the forest remains of the National Monument of Pontoes Capixabas, because of higher species richness and diversity. Furthermore, they indicate the epidemiological role of Lu. longipalpis as the vector of Leishmania infantum within the study area, and the no evident role of Mg. migonei.

Introduction and objectives: Jorge Lobo’s disease is a rare chronic disease caused by the fungus Lacazia loboi, characterized by keloid lesions on the skin. The objective is to perform a review of the literature and report the first case of Jorge Lobo’s disease treated at the University Hospital Cassiano Antonio Moraes (HUCAM). Materials and methods: The study data were obtained retrospectively from the analysis and systematic review of the medical records of a patient with Jorge Lobo’s disease identified in HUCAM. Case Report: A male patient aged 37 years was referred to HUCAM in April 2010 presenting with keloid lesions and a previous diagnosis of lobomycosis by histopathology examination. On December 2011 a partial removal of the lesions with grafting was performed. Three months later, there was recurrence of lobomycosis on the grafted tissue. In June 2012 drug treatment with clofazimine associated with fluconazole was started. The lack of satisfactory clinical response after two months meant that the disease was considered beyond cure. Discussion and literature review: lobomycosis is an endemic disease in the brazilian Amazon and other humid tropical areas of Central and South America, and its transmission occurs by inoculation of the fungus through breaks in the skin. Diagnosis is made by clinical examination and histopathologiy or cytology The choice of therapy varies with the extent of lesions. However, none of the therapies described in the literature is fully effective. Conclusion: In this case, the two main therapeutic approaches failed, and the disease was considered incurable.

This is the first record of Evandromyia (Aldamyia) sericea in southeastern Brazil and report the southernmost record ever of this species in the world. Specimens were collected in the Reserva Biologica de Duas Bocas, municipality of Cariacica, state of the Espirito Santo. Captures of the sand flies were carried out monthly from April to June 2008 during four consecutives nights using two automatic light traps (CDC model) and a modified Shannon trap for each night. The finding of this species corroborates the hypothesis of the phylogeographical proximity of the Amazon Rainforest and the Atlantic Forest.

Taxonomic studies of leishmanial isolates fromthe New World indicate tremendous diversitywithin this genus (E Cupolillo et al. 1994 Am JTrop Med Hyg 50: 296-311). A number of newLeishmania species from sylvan areas of theNeotropics have been described recently. Some ofthose parasites are associated with disease in hu-mans; others appear to be restricted to lower or-ders of mammals, such as rodents and edentates(G Grimaldi et al. 1989 Am J Trop Med Hyg 41:687-725). However, some of the latter parasitesmay yet be shown capable of causing human dis-ease, particularly in persons with altered cellularimmune responses.During a survey of potential reservoir hosts ofLeishmania done in the State of Espirito Santo, Bra-zil (1982-1985), three leishmanial parasites (des-ignated with stock codes MPRO/BR/82/RV203,MPRO/BR/82/RV228, and MPRO/BR/85/RV260)were isolated from skin lesion samples taken fromspiny-rats, Proechimys iheringi (Rodentia,Echimydae), captured in a secondary forest locatedin the municipality of Viana (A Falqueto et al. 1985Mem Inst Oswaldo Cruz 80: 497). In studies us-ing an indirect radioimmune binding assay (RIA)and a large panel of monoclonal antibodies (Mabs)derived for selected species of Leishmania , the iso-lates were characterized as members of the L.mexicana complex. These strains were differentfrom those species infective to humans, in that theyreacted with either the L.(L.) amazonensis or L.(L.) mexicana specific MAbs. Furthermore, thiswas also the case with cloned organisms from twostocks showing that they were not mixed popula-tions of Leishmania (G Grimaldi et al. 1987 Am JTrop Med Hyg 36: 270-287). This group of dis-tinct parasites presented, however, similarserodeme patterns in relation to the new recentlydescribed species of the Leishmania subgenus, L.(L.) forattinii (E Yoshida et al. 1993 Mem InstOswaldo Cruz 88: 397-406).The latter parasite was isolated from sylvaticreservoir hosts (Didelphis marsupialis aurita andP. iheringi denigratus ) captured on regions endemicfor American cutaneous leishmaniasis in Brazil,respectively in the municipalities of Conchas, SaoPaulo (E Yoshida et al. 1979 Rev Inst Med Trop SPaulo 21: 110-113) and Tres Bracos, Bahia (ABarreto et al. 1985 Rev Soc Bras Med Trop 18:243-246). The general morphology and the growthcharacteristics in vitro of the new species were simi-lar to those of other L. mexicana complex para-sites. Here we have further studied the Espirito Santoisolates using additional techniques for the char-acterization of Leishmania . The molecular proce-dures used for typing the strains (isoenzyme elec-trophoresis, blot enzyme binding assay usingMAbs, restriction endonuclease fragment patternsof k-DNA, and molecular karyotype analysis) havebeen described in detail in previous publications(Yoshida et al. 1979, 1983 loc. cit. , Cupolillo et al.1994 loc. cit., A Franco et al. 1997 Mem InstOswaldo Cruz 92: 63-68).Results of electrophoretic analysis were basedon the following enzymatic loci, namely: glucose-6-phosphate dehydrogenase (G6PDH), malate de-hydrogenase (MDH), isocitrate dehydrogenase(IDHNADP), 6-phosphogluconate dehydrogenase(6PGDH), nucleotidase (NH1 and NH2), glucosephosphate isomerase (GPI), phosphoglucomutase(PGM), proline dipeptidase (PEPD), leucine pep-tidase (PEP2), malic enzyme (ME), mannose phos-phate isomerase (MPI), and aconitate hydratase(ACON). The enzyme profiles of the selected iso-lated signature MPRO/BR/82/RV228, which rep-resents the poorly defined group of parasites from

This study compares the distribution of anopheline mosquitoes in a malaria-endemic municipality (MAL) and a malaria-free municipality (FREE) in an area of the Atlantic Forest in Brazil. Simultaneous quarterly nightly captures were made in three locations in each municipality. One Shannon light trap (Shannon light traps were home made according to specifications published in Am. J. Trop. Med. 1939; 19: 131-140) (SLT) and five CDC light traps (a kind of automatic trap fed by batteries of 12 V and 7 amp/h, with dry ice as a source of CO2; John W. Hock Company, Gainesville, FL) (CLT) (two in the canopy and three at ground level) were operated from 6:00 p.m. to 6:00 a.m. More specimens were captured in MAL (362 in SLTs and 126 in CLTs) than in FREE (66 in SLTs and 59 in CLTs). For the SLTs, Simpson's dominance index was similar in MAL and FREE (D = 0.15 versus D = 0.203, P0.7), but Shannon's diversity index was higher for MAL = 1.969 versus H = 1.641, P0.01). For the CLTs, Simpson's dominance index was higher in MAL (D = 0.416 versus 0.2688, P0.001), and the Shannon diversity index was higher in FREE (H = 1.5222 versus H = 1.115, P0.01). In SLTs, Anopheles (Kerteszia) cruzii s.l. frequencies were higher in MAL (chi2 = 23.39; P = 0.000001). In CLTs, An. cruzii s.l. was present in all strata in MAL but only in the canopy inside the forest in FREE (17 specimens). An. cruzii s.l. represented a higher proportion of anophelines in MAL (chi2 = 31; P0.000001). The factors that differed in these two areas were anopheline species density and An. cruzii s.l. abundance and distribution.

Two new species of Sycorax from the Atlantic Rain Forest of Espírito Santo, Sycorax canaanensis Santos, BravoFalqueto sp. nov. and Sycorax tuberculata Santos, BravoFalqueto sp. nov. are described and illustrated. Male speci-mens were collected with CDC light traps in the Biological Reserve of Augusto Ruschi, municipality of Santa Teresa, state of Espírito Santo, Brazil. This finding raises the number of described Western Hemisphere Sycorax species to 15.

The occurrence of American cutaneous leishmaniasis (ACL) in areas modified by humans indicates that phlebotomine sand fly vectors breed close to human habitations. Potential peridomiciliary breeding sites of phlebotomines were sampled in an area of transmission of Leishmania (Viannia) braziliensis in Southeastern Brazil. Three concentric circles rounding houses and domestic animal shelters, with radii of 20, 40, and 60 m, defined the area to be monitored using adult emergence traps. Of the 67 phlebotomines collected, Lutzomyia intermedia comprised 71.6%; Lutzomyia schreiberi, 20.9%; and Lutzomyia migonei, 4.5%. The predominance of L. intermedia, the main species suspected of transmitting L. (V.) braziliensis in Southeastern Brazil, indicates its participation in the domiciliary transmission of ACL, providing evidence that the domiciliary ACL transmission cycle might be maintained by phlebotomines that breed close to human habitations. This finding might also help in planning measures that would make the peridomiciliary environment less favorable for phlebotomine breeding sites.

The aim of this study was to evaluate the sand fly fauna of American visceral leishmaniasis (AVL) endemic areas within the Central Atlantic Forest Biodiversity Corridor, State of Espírito Santo, southeastern Brazil. The sand fly captures were performed between January, 1989 and December, 2003 in localities where autochthonous cases of AVL were recorded, as well as in their boundary areas. Sand flies were collected from surrounding houses and domestic animal shelters using two to five CDC automatic light traps, and manual captures were also performed using mouth aspirators in one illuminated Shannon trap during the first four hours of the night. We used cladistic analysis to determine the geographic relationships among the collected sand fly species as well as the index species for the occurrence of other sand flies. A total of 62,469 sand flies belonging to 17 species and eight genera was collected in 164 localities from nine municipalities with AVL records. The richness (S=17) and diversity (H=0.971) of sand flies were lower than in conservation areas and similar to modified environments in the Atlantic Forest of Espírito Santo. Lutzomyia longipalpis was identified in 79 localities. The cladistic analysis identified Evandromyia lenti as the index species for Lutzomyia longipalpis. The latter seems to be the main vector of AVL in the Central Atlantic Forest Biodiversity Corridor due to its high abundance and distribution matching the disease occurrence. Therefore, Evandromyia lenti may be used as an index species for the occurrence of Lutzomyia longipalpis.

Em área endêmica de leishmaniose tegumentar no Município de Santa Leopoldina, Espírito Santo, foi realizada uma tentativa de controle da endemia por meio do tratamento dos cães doentes. Como controle foi utilizada uma área com características semelhantes no Município de Afonso Cláudio, onde os cães permaneceram sem tratamento. A avaliação inicial revelou que 34 (24%) dos 141 moradores da primeira área eram reatores ao teste intradérmico de Montenegro; entre os cães, 8 (30,7%), dos 26 existentes, apresentavam leishmaniose. Na área controle, 37 (29%), dos 127 moradores, eram positivos ao teste de Montenegro, enquanto 7 (16%), dos 44 cães, estavam doentes. Após o tratamento dos cães doentes na primeira área, as duas populações foram acompanhadas durante dois anos. A eficácia da medida de controle foi avaliada por meio da incidência de novas infecções humanas. Apesar de o número de novos infectados ter sido superior na área-controle, a análise estatística revelou que a diferença não foi significativa. Fatores diversos, tais como o tamanho das amostras estudadas, o método de diagnóstico da leishmaniose canina e a dificuldade de controle dos novos cães introduzidos na área, teriam influenciado negativamente nos resultados do experimento. An attempt was made to control cutaneous leishmaniasis in the municipality of Santa Leopoldina, Espírito Santo State, Brazil, by treating infected dogs. Another area in the municipality of Afonso Cláudio, which is ecologically similar, served as the control area, and dogs were left untreated there. In an initial survey, 34/141 inhabitants of the first area had positive leishmanin tests and 8/26 dogs had cutaneous leishmaniasis, while in the second area 37/127 individuals had positive leishmanin tests and 7/44 dogs were infected. After treatment of the sick dogs in the first area the populations of both areas were periodically examined for new infections. More new human infections were observed in the control area, but the difference was not statistically significant.

Lutzomyia longipalpis (Diptera: Psychodidae) is the principal vector of American visceral leishmaniasis. Several studies have indicated that the Lu. longipalpis population structure is complex. It has been suggested that genetic divergence caused by genetic drift, selection, or both may affect the vectorial capacity of Lu. longipalpis. However, it remains unclear whether genetic differences among Lu. longipalpis populations are directly implicated in the transmission features of visceral leishmaniasis. We evaluated the genetic composition and the patterns of genetic differentiation among Lu. longipalpis populations collected from regions with different patterns of transmission of visceral leishmaniasis by analyzing the sequence variation in the mitochondrial cytochrome b gene. Furthermore, we investigated the temporal distribution of haplotypes and compared our results with those obtained in a previous study. Our data indicate that there are differences in the haplotype composition and that there has been significant differentiation between the analyzed populations. Our results reveal that measures used to control visceral leishmaniasis might have influenced the genetic composition of the vector population. This finding raises important questions concerning the epidemiology of visceral leishmaniasis, because these differences in the genetic structures among populations of Lu. longipalpis may have implications with respect to their efficiency as vectors for visceral leishmaniasis.

INTRODUCTION: The article describes the epidemiologic profile of dengue cases in Vitória, the capital of Espírito Santo, Brazil, from 2000 to 2009, aimed at identifying risk groups regarding the incidence and severity of the disease. METHODS: Confirmed cases of dengue among city residents during ten years were classified as dengue fever, dengue hemorrhagic fever, dengue shock syndrome and dengue with complications, and analyzed according to sex, age, race-color and education. RESULTS: The proportion of dengue cases was highest among women aged 20 to 29 years-old and similar between whites and blacks. A gradual decrease occurred in the percentage of dengue cases in the population aged 15 years-old or more, in the historical series of 10 years, and a growing increase in individuals less than 15 years-old, showing statistical significance. The fatality rate ranged from zero to 0.3% for all forms of dengue and from 0.2% to 18.2% for severe forms. CONCLUSIONS: The profile of those affected by the disease in the municipality is similar to those affected in Brazil. The increasing number of cases in individuals under 15 years-old corroborates the results of recent studies in other Brazilian municipalities. INTRODUÇÃO: O artigo descreve o perfil epidemiológico dos casos de dengue ocorridos no município de Vitória, capital do Estado do Espírito Santo, no período de 2000 a 2009, procurando identificar grupos de risco para a incidência e gravidade da doença. MÉTODOS: Os casos de dengue, confirmados em residentes do município no período de dez anos, foram classificados como febre da dengue, febre hemorrágica da dengue, síndrome do choque da dengue e dengue com complicações, e analisados segundo sexo, faixa etária, raça-cor e escolaridade. RESULTADOS: A proporção de casos de dengue foi maior entre mulheres, indivíduos de 20 a 29 anos, e semelhante entre brancos e afrodescendentes. Houve uma diminuição gradual do percentual de casos de dengue na população com 15 anos ou mais, na série histórica de 10 anos, e um aumento crescente em menores de 15 anos de idade, com significância estatística. A taxa de letalidade variou de zero a 0,3% para todas as formas de dengue, e de 0,2% a 18,2% para as formas graves. CONCLUSÕES: O perfil dos acometidos pela dengue no município é semelhante ao do Brasil. O aumento crescente de casos em menores de 15 anos corrobora os resultados de estudos recentes em outros municípios brasileiros.

Synthesis of the studies about the transmission cycle of malaria in an area of very low incidence outside the Amazon Region in Brazil Crispim Cerutti Jr, Aloisio Falqueto, Helder Ricas Rezende, Renata Soares, Isabel Alves, Delsio Natal, Paulo Roberto Urbinatti, Tasciane Yamasaki, Ana Maria Ribeiro de Castro Duarte, Rosely dos Santos Malafronte From 16 International Symposium on HIV and Emerging Infectious Diseases Marseille, France. 24-26 March 2010

The occurrence of American visceral leishmaniasis in the State of Espírito Santo, Brazil has always been associated with the presence of the Lutzomyia longipalpis vector. The geographic distribution of this vector in this state is related to the presence of specific geoclimatic characteristics, such as a dry climate, low elevations (< 450m), steep slopes and rocky outcrops. The occurrence of human autochthonous cases of American visceral leishmaniasis in municipalities without these geoclimatic characteristics justifies the present study and our main goal was to test the association between the occurrence of American visceral leishmaniasis and the presence of the Lu. longipalpis. Sand flies were captured monthly from July 2006 to August 2007 using Shannon and CDC traps in two municipalities with records of autochthonous American visceral leishmaniasis and one with no record. We captured 13,112 sand flies, but no Lu. longipalpis was found. The absence of Lu. longipalpis and the possible role of another American visceral leishmaniasis vector in these localities were discussed. A ocorrência de leishmaniose visceral americana no Estado do Espírito Santo, Brasil, sempre foi associada à presença do vetor Lutzomyia longipalpis. A distribuição geográfica desse vetor no estado está relacionada à ocorrência de características geoclimáticas específicas, tais como clima seco, baixas altitudes, relevo acidentado e afloramentos rochosos A ocorrência de casos humanos autóctones de leishmaniose visceral americana em municípios sem essas características geoclimáticas justificou o presente estudo, com objetivo de testar a associação entre a ocorrência de leishmaniose visceral americana e a presença de Lu. longipalpis. Flebotomíneos foram capturados mensalmente de julho de 2006 a agosto de 2007 com uso de armadilhas de Shannon e CDC em dois municípios com registros de leishmaniose visceral americana autóctone e um sem registros. Foram capturados 13.112 flebotomíneos, mas nenhum Lu. longipalpis foi encontrado. A ausência de Lu. longipalpis e a possível participação de outro vetor de leishmaniose visceral americana nessas localidades foram discutidas.

In this study, we sought to identify sand fly vectors of the Leishmania species that circulate in distinct eco-epidemiological disease-endemic rural areas within the Espirito Santo State in southeastern Brazil. PCR amplification of a conserved region of the minicircle kDNA was used to estimate infection rates in field-captured, peridomestic female sand flies. Only 13 of the 1689 female sand fly specimens (0.77%) actually contained Leishmania DNA. Leishmania braziliensis infections were found in Lutzomyia intermedia and Lu. whitmani, and, for the first time, in Lu. fischeri and Lu. ferreirana. Interestingly, the high rate of genetic polymorphism of the L. braziliensis parasites in one of the disease-endemic areas that were studied may reflect specific transmission cycles involving different sand fly vectors.

A construção de represas geralmente causa modificações na composição da fauna em sua área de influência. Na área de implantação da Usina Hidrelétrica de Rosal, Estados do Espírito Santo e Rio de Janeiro, foram avaliadas modificações na abundância de anofelinos, planorbídeos e flebotomíneos antes (1998-2000) e após (2000-2005) o represamento. Foram definidos nove pontos de coleta, cada qual representado por uma moradia e seus anexos, abrigos de animais domésticos e coleções hídricas num raio de 150m. Coletaram-se 103 anofelinos adultos antes do represamento e 313 depois, 200 imaturos antes e 708 depois, 868 planorbídeos antes e 486 depois, e 2.979 flebotomíneos antes e 912 depois. O registro de vetores dentre anofelinos, planorbídeos e flebotomíneos revela o potencial da área para transmissão de malária, esquistossomose e leishmaniose tegumentar. As transformações ambientais ocorridas, no entanto, não aumentaram o risco para ocorrência dessas doenças. The construction of dams usually changes the composition of the fauna within their areas of influence. In the area where the Rosal hydroelectric power station has been installed, in the States of Espírito Santo and Rio de Janeiro, changes to the abundance of anophelinae, planorbidae and phlebotominae were evaluated before (1998-2000) and after (2000-2005) the damming. Nine sample collection points were defined, each represented by a home and its outbuildings, shelters for domestic animals and water collection points within a radius of 150m. One hundred and three adult anophelinae were collected before the damming and 313 afterwards; 200 immature anophelinae before and 708 afterwards; 868 planorbidae before and 486 afterwards; and 2,979 phlebotominae before and 912 afterwards. The vector record, consisting of anophelinae, planorbidae and phlebotominae, shows the potential of this area for transmission of malaria, schistosomiasis and American cutaneous leishmaniasis. However, the environmental changes did not increase the risk of the occurrence of these diseases.

The effect of deltamethrin of the sandfly population in a focus of American Cutaneous Leishmaniasis in Viana, Espírito Santo State is described. The phlebotomine population density was determined inside and outside houses during a one year period in a treated and untreated area. The results showed a significant reduction in the number of sandflies inside houses in the treated area compared with both the untreated area and the same area before spraying. Despite having a residual action for 12 months after spraying the insecticide was ineffective outside houses. It is concluded therefore that deltamethrin is an efficient insecticide for reducing the number of phlebotomine sandflies inside houses.

American tegumentary leishmaniasis is endemic to the Espírito Santo State, Brazil, where it is widely distributed. The composition of the phlebotomine sand fly fauna in an American tegumentary leishmaniasis focus was determined by monthly sampling, using Shannon light traps in an Atlantic Forest reserve and adjacent habitat that had been modified by human activity. Seasonal fluctuations in numbers of the most abundant species were also monitored from June 2004 to May 2006. Of the 6,176 specimens collected, 47.4% were captured in the forest and 52.6% in the disturbed habitat. Although Lutzomyia davisi (60.8%) predominated in specimens from the forest, those captured near human dwellings consisted almost entirely of Lu. choti (72%) and Lu. intermedia (24.3%). All three species occurred throughout the year. Based on our findings, Lu. intermedia probably acts as the principal domestic Leishmania vector in the study area.

Estudo caso controle que objetivou identificar fatores individuais de risco relacionados à transmissão da doença. O grupo caso, composto por 90 pacientes de hanseníase notificados no SINAN entre 2003 e 2006; o grupo controle, constituído por 270 indivíduos sadios, pareados por sexo e faixa etária. Houve associação significativa entre a ocorrência da doença e a presença atual (OR= 2,9) e antiga (OR=5,0) de hanseníase entre parentes co-sangüíneos. Sabendo-se que o exame único dos contatos, no ato do diagnóstico, detecta uma parcela mínima dos casos, propõe-se a realização de exames periódicos dos contatos de hansenianos a fim de detectar os novos casos. This case-control study aimed to identify individual risk factors regarding the transmission of leprosy. The group case represented by 90 cases of leprosy sick people pontificated at SINAN during 2003 and 2006 and a group control constituted by 270 healthy people, paired by gender and age. There was significant statistical between occurrence of the disease and its current presence (OR: 2,9) and old cases (OR= 5,0) of leprosy among co-sanguine relatives. Knowing that the only exam of the contact in the act of the diagnostic detects a minimum part of the new cases, it is proposed to execute periodical examinations of the contact of leprosy for detecting the new cases. Estudio caso controle que objetivó identificar factores individuales del riesgo relacionados a transmisión de la enfermedad. El grupo caso, compuesto por 90 pacientes con lepra notificados en el SINAN entre 2003 y 2006; y el grupo controle, constituido por 270 individuos saludables, pareados por sexo y edad. Hubo asociación significativa entre la ocurrencia de la enfermedad y la presencia actual (OR= 2,9) y antigua (OR=5,0) de la lepra entre parientes co-sanguíneos. Sabiéndose que el examen único de los contactos en el acto del diagnóstico detecta una parcela mínima de los casos, se propone a la realización del examen periódico de los contactos de leprosos a fin de detectar los nuevos casos.

In Brazil, introduced malaria occurs from the flat to the sloping hot areas, predominantly outside the Amazon Region, where endemic malaria has occurred in the past. This is a consequence of human migrations to other Brazilian states, including the state of Espírito Santo (ES). The objective of this study was to use geoprocessing to define the areas at risk of introduced malaria transmission and evaluate the vectorial importance of species of anophelines in ES. Anophelines were sampled from 1997-2005 in 297 rural localities identified or not identified as foci of malaria during the last 20 years. The geoclimatic variables temperature, relief and marine influence were obtained from a database of the ES Natural Units. The 14,663 anophelines captured belonged to 22 species. A significant association was found between the occurrence of malaria foci and the presence of hot, low-lying areas or gently undulating to undulating relief. The occurrence of the disease was associated with the presence of Anopheles darlingi and Anopheles aquasalis. Geoprocessing was determined to be a useful tool for defining areas at risk for malaria and vectors in ES.

The construction of dams usually changes the composition of the fauna within their areas of influence. In the area where the Rosal hydroelectric power station has been installed, in the States of Espírito Santo and Rio de Janeiro, changes to the abundance of anophelinae, planorbidae and phlebotominae were evaluated before (1998-2000) and after (2000-2005) the damming. Nine sample collection points were defined, each represented by a home and its outbuildings, shelters for domestic animals and water collection points within a radius of 150m. One hundred and three adult anophelinae were collected before the damming and 313 afterwards; 200 immature anophelinae before and 708 afterwards; 868 planorbidae before and 486 afterwards; and 2,979 phlebotominae before and 912 afterwards. The vector record, consisting of anophelinae, planorbidae and phlebotominae, shows the potential of this area for transmission of malaria, schistosomiasis and American cutaneous leishmaniasis. However, the environmental changes did not increase the risk of the occurrence of these diseases.

This case-control study aimed to identify individual risk factors regarding the transmission of leprosy. The group case represented by 90 cases of leprosy sick people pontificated at SINAN during 2003 and 2006 and a group control constituted by 270 healthy people, paired by gender and age. There was significant statistical between occurrence of the disease and its current presence (OR: 2,9) and old cases (OR= 5,0) of leprosy among co-sanguine relatives. Knowing that the only exam of the contact in the act of the diagnostic detects a minimum part of the new cases, it is proposed to execute periodical examinations of the contact of leprosy for detecting the new cases.

Os flebotomíneos Nyssomyia yuilli yuilli (Young & Porter) (Diptera: Psychodidae) e Trichopygomyia longispina (Mangabeira) (Diptera: Psychodidae) são registrados pela primeira vez no estado do Espírito Santo, Brasil. The phlebotomine sand flies Nyssomyia yuilli yuilli (Young & Porter) (Diptera: Psychodidae) and Trichopygomyia longispina (Mangabeira) (Diptera: Psychodidae) are recorded in the state of Espírito Santo, Brazil for the first time.

Autochthonous malaria cases in the state of Espírito Santo, Brazil, are distributed in mountainous regions surrounded by the Atlantic Forest. While some aspects of this disease are unclear, detection of possible vector species can help to elucidate epidemiological uncertainties. Entomological and natural infection studies were carried out using anophelines (Diptera: Culicidae) captured in the municipality of Santa Tereza, ES. Monthly captures were made from March 2004 to February 2006. CDC-CO2 traps were used from dusk (6:00 P.M.) to dawn (6:00 A.M.) to capture anophelines in the following habitats: near the houses, in open areas (at ground level) and inside, and at the margins of the forest (canopy and ground level). Shannon light traps were also used at the same locations of the CDC-CO2 traps. A total of 2,290 anophelines within 10 species were captured. The relative frequency of Anopheles (Kerteszia) cruzii Dyar & Knab / A.(K.) homunculus Komp was the highest, with the majority captured in CDC-CO2 traps installed in the forest canopy. The main species captured in Shannon traps was A.(Nyssorhynchus) strodei Root. The largest number of anophelines was captured from July to September and from 6:00 P.M. to 10:00 P.M. Anopheles (K.) cruzii is the probable vector for malaria transmission inside or near the Atlantic Forest fragments, but the role of other species cannot be ignored, as 53% of the sampled anophelines belonged to the subgenus Nyssorhynchus. The natural infection of A. cruzii, A. parvus (Chagas) and A. galvaoi Causey, Deane & Deane by Plasmodium vivax detected by PCR from DNA extracted from their thoraxes supports this view.

Background Extra-Amazonian autochthonous Plasmodium vivax infections have been reported in mountainous regions surrounded by the Atlantic Forest in Espírito Santo state, Brazil. Methods Sixty-five patients and 1,777 residents were surveyed between April 2001 and March 2004. Laboratory methods included thin and thick smears, multiplex-PCR, immunofluorescent assay (IFA) against P. vivax and Plasmodium malariae crude blood-stage antigens and enzyme-linked immunosorbent assay (ELISA) for antibodies against the P. vivax-complex (P. vivax and variants) and P. malariae/Plasmodium brasilianum circumsporozoite-protein (CSP) antigens. Results Average patient age was 35.1 years. Most (78.5%) were males; 64.6% lived in rural areas; 35.4% were farmers; and 12.3% students. There was no relevant history of travel. Ninety-five per cent of the patients were experiencing their first episode of malaria. Laboratory data from 51 patients were consistent with P. vivax infection, which was determined by thin smear. Of these samples, 48 were assayed by multiplex-PCR. Forty-five were positive for P. vivax, confirming the parasitological results, while P. malariae was detected in one sample and two gave negative results. Fifty percent of the 50 patients tested had IgG antibodies against the P. vivax-complex or P. malariae CSP as determined by ELISA. The percentages of residents with IgM and IgG antibodies detected by IFA for P. malariae, P. vivax and Plasmodium falciparum who did not complain of malaria symptoms at the time blood was collected were 30.1% and 56.5%, 6.2% and 37.7%, and 13.5% and 13%, respectively. The same sera that reacted to P. vivax also reacted to P. malariae. The following numbers of samples were positive in multiplex-PCR: 23 for P. vivax; 15 for P. malariae; 9 for P. falciparum and only one for P. falciparum and P. malariae. All thin and thick smears were negative. ELISA against CSP antigens was positive in 25.4%, 6.3%, 10.7% and 15.1% of the samples tested for "classical" P. vivax (VK210), VK247, P. vivax-like and P. malariae, respectively. Anopheline captures in the transmission area revealed only zoophilic and exophilic species. Conclusion The low incidence of malaria cases, the finding of asymptomatic inhabitants and the geographic separation of patients allied to serological and molecular results raise the possibility of the existence of a simian reservoir in these areas.

DNA barcoding has been an effective tool for species identification in several animal groups. Here, we used DNA barcoding to discriminate between 47 morphologically distinct species of Brazilian sand flies. DNA barcodes correctly identified approximately 90% of the sampled taxa (42 morphologically distinct species) using clustering based on neighbor-joining distance, of which four species showed comparatively higher maximum values of divergence (range 4.23-19.04%), indicating cryptic diversity. The DNA barcodes also corroborated the resurrection of two species within the shannoni complex and provided an efficient tool to differentiate between morphologically indistinguishable females of closely related species. Taken together, our results validate the effectiveness of DNA barcoding for species identification and the discovery of cryptic diversity in sand flies from Brazil.

Some insects that are vectors of human diseases have accompanied man in his migrations throughout the world and breed exclusively in the proximity of human dwellings. The mosquito Aedes aegypti has been responsible for epidemics of dengue in Brazil and its presence also constitutes a serious risk for future outbreaks of urban yellow fever. The failure of campaigns to eradicate this species justifies the search for alternative breeding sites, which may be beyond the reach of present control measures. In this study the occurrence of Aedes aegypti breeding sites in native bromeliads on rocky slopes was investigated in five areas of Vitória, capital of the Brazilian State of Espírito Santo, ES. Water contained in the bromeliads was collected with the aid of a suction apparatus to search for culicid larvae. The degree of infestation of buildings in adjacent urban areas was evaluated simultaneously. Culicid larvae were found in bromeliads in four of the five areas investigated, Aedes aegypti being present in two areas. The presence of breeding sites in bromeliads was not related to indices of infestation of buildings in adjacent areas. Further studies are necessary to define whether breeding sites in bromeliads constitute primary foci of Aedes aegypti, or are a consequence of high infestation levels in urban areas.