Relationship between vertical stratification and feeding habits of mosquito (Diptera: Culicidae) assemblages collected in conservation units in the green belt of the city of São Paulo, Brazil.

São Paulo is one of the largest cities in the world and has several characteristics that favor a diversity of urban and wild mosquitoes. Little is known about how variations in mosquito diversity and feeding preferences for different hosts in different vegetation strata can influence the risk of pathogen transmission to humans. We investigated vertical stratification of mosquitoes and its relationship with vertebrate hosts in environments with different degrees of conservation in two conservation units in the city of São Paulo. Adult mosquitoes were collected using CDC traps, aspiration and Shannon traps. After morphological identification, host blood in engorged females was analyzed by PCR with a vertebrate-specific primer set based on mitochondrial cytochrome b DNA of vertebrates commonly found in the two conservation units. Although a higher abundance of the species Anopheles cruzii and Culex nigripalpus was found in the canopy, blood not only from birds but also from humans and rodents was identified in these mosquitoes. In one of the units, Wyeomyia confusa and Limatus durhamii were found occupying mainly niches at ground level while Culex vaxus was frequently found in the canopy. Haemagogus leucocelaenus, the main vector of yellow fever, was found in low abundance at all collection points, particularly in the canopy. Species richness and composition tended to vary little between canopy and ground level in the same environment, but the abundance between canopy and ground level varied more depending on the species analyzed, the most abundant and frequent species exhibiting a predilection for the canopy. Even those mosquito species observed more frequently in the canopy did not show an association with hosts found in this stratum as most of the blood identified in these species was from humans, suggesting opportunist feeding behavior, i.e., feeding on the most readily available host in the environment. The two most common species in the study, An. cruzii and Cx. nigripalpus, may be able to act as bridge vectors for pathogens to circulate between the forest canopy and ground level.

Phenotypic and genetic variation of Triatoma costalimai (Hemiptera: Reduviidae)

Abstract INTRODUCTION: We aimed to study intraspecific variation in Triatoma costalimai, a potential vector of Chagas disease present in Brazil and Bolivia. METHODS: We analyzed phenotypic (connexivum color patterns, wing morphometrics) and genetic variation (16S mtDNA) of three Brazilian T. costalimai populations. We compared 16S sequences with those of putative Bolivian T. costalimai and its sister species, T. jatai. RESULTS: Brazilian populations had different connexivum color patterns and forewing shapes. A 16S mtDNA haplotype network showed a clear separation of Brazilian T. costalimai from both T. jatai and Bolivian T. costalimai. CONCLUSIONS: We report considerable variability in T. costalimai populations.

Phenotypic and genetic variation of Triatoma costalimai (Hemiptera: Reduviidae).

INTRODUCTION: We aimed to study intraspecific variation in Triatoma costalimai, a potential vector of Chagas disease present in Brazil and Bolivia. METHODS: We analyzed phenotypic (connexivum color patterns, wing morphometrics) and genetic variation (16S mtDNA) of three Brazilian T. costalimai populations. We compared 16S sequences with those of putative Bolivian T. costalimai and its sister species, T. jatai. RESULTS: Brazilian populations had different connexivum color patterns and forewing shapes. A 16S mtDNA haplotype network showed a clear separation of Brazilian T. costalimai from both T. jatai and Bolivian T. costalimai. CONCLUSIONS: We report considerable variability in T. costalimai populations.

Morphological study of the urotergite I process in ten species of the genus Triatoma (Hemiptera, Reduviidae, Triatominae).

The insects of subfamily Triatominae exhibit many morphological features used for taxonomic identification. In some species, however, these features are very similar. The authors have proposed by first time the description of the structure referred as urotergite I process. The objective of the study was analyse the use of urotergite I process in the species of the genus Triatoma as a taxonomic feature that may contribute to the more precise and correct identification of these Trypanosoma cruzi Chagas, 1909 vectors. Scanning electron microscopy (SEM) was used to describe the structure of urotergite I process in ten Triatoma (Hemiptera, Reduviidae, Triatominae) species: Triatoma brasiliensis Neiva, 1911; Triatoma circummaculata Stål, 1859; Triatoma infestans (Klug, 1834); Triatoma maculata (Erichson, 1848); Triatoma pseudomaculata Corrêa & Espínola, 1964; Triatoma rubrovaria (Blanchard, 1843); Triatoma sordida (Stål, 1859); Triatoma tibiamaculata (Pinto, 1926); Triatoma vitticeps (Stål, 1859) and Triatoma wygodzinskyi Lent, 1951. The morphological description of the urotergite I process reflects the taxonomic value of the structure for separating and identifying species of ten specimens of the genus Triatoma. The morphological pattern of the urotergite I process on these ten species was compared, and interspecific variability was observed. We suggest the use of the urotergite I process as a complementary character to identify insects of the subfamily Triatominae. In the future, the use of the urotergite I process in dichotomic keys to identify triatomines may contribute to the improvement of the entomological surveillance of Chagas' disease.

Occurrences of triatomines (Hemiptera: Reduviidae) and first reports of Panstrongylus geniculatus in urban environments in the city of Sao Paulo, Brazil.

This note reports on occurrences of triatomine species in the city of Sao Paulo, Brazil, registered between 1988 and 2017. Records of triatomines captured in Sao Paulo are based on specimens received spontaneously from Health Surveillance Centers, Health Centers and Zoonosis Control Centers in the city as well as from citizens. Species were identified morphologically at the Public Health Entomology Laboratory, Faculty of Public Health, University of Sao Paulo, where the triatomines, which are vectors of Chagas disease, were tested for Trypanosoma cruzi infection. The first reported occurrence of triatomine bugs in urban Sao Paulo was in 1988. The specimen, which was captured in Jardim Sao Luiz district, was from the genus Panstrongylus and was registered as Panstrongylus sp. but was not sexed. Since this first recorded occurrence, the following species have been found in the city: Panstrongylus geniculatus (2 occurrences), P. megistus (15 occurrences), Triatoma infestans (1 occurrence) and T. sordida (3 occurrences). In this paper, the importance of reporting occurrences of triatomine bugs in the city of Sao Paulo, one of the largest metropolis in the world, is discussed with an emphasis on P. megistus. The occurrences discussed here indicate the importance of entomological surveillance for these vectors even in urban centers although the possibility of vector transmission of Chagas disease in these centers is very low.

Catalog of the entomologic collections of the Faculty of Public Health of the University of Sao Paulo - (2nd series ii): Triatominae (Hemiptera, Reduviidae).

This article reports a list with 912 specimens of the subfamily Triatominae deposited in the Entomological Collection of the Faculty of Public Health of the University of Sao Paulo. The collection is composed of 1 holotype, 3 alotypes, 15 paralectotypes, 77 paratypes, distributed in 5 tribes and 12 genera: Tribus Alberprosenini: genus Alberprosenia Martinez & Carcavallo, 1977; Tribus Bolboderini: genus Microtriatoma Prosen & Martinez, 1952; Tribus Cavernicolini: genus Cavernicola Barber, 1937; Tribus Rhodnini: genus Psammolestes Bergroth, 1941; genus Rhodnius Stal, 1859; Tribus Triatomini: genus Dipetalogaster Usinger 1939; genus Eratyrus Stal 1859; genus Hermanlentia Jurberg & Galvão, 1997; genus Linshcosteus Distant, 1904; 1944; genus Panstrongylus Berg 1879; genus Paratriatoma Barber 1938; genus Triatoma Laporte 1833.

Wing variation in Culex nigripalpus (Diptera: Culicidae) in urban parks.

BACKGROUND: Culex nigripalpus has a wide geographical distribution and is found in North and South America. Females are considered primary vectors for several arboviruses, including Saint Louis encephalitis virus, Venezuelan equine encephalitis virus and Eastern equine encephalitis virus, as well as a potential vector of West Nile virus. In view of the epidemiological importance of this mosquito and its high abundance, this study sought to investigate wing variation in Cx. nigripalpus populations from urban parks in the city of São Paulo, Brazil. METHODS: Female mosquitoes were collected in seven urban parks in the city of São Paulo between 2011 and 2013. Eighteen landmark coordinates from the right wing of each female mosquito were digitized, and the dissimilarities between populations were assessed by canonical variate analysis and cross-validated reclassification and by constructing a Neighbor-Joining (NJ) tree based on Mahalanobis distances. The centroid size was calculated to determine mean wing size in each population. RESULTS: Canonical variate analysis based on fixed landmarks of the wing revealed a pattern of segregation between urban and sylvatic Cx. nigripalpus, a similar result to that revealed by the NJ tree topology, in which the population from Shangrilá Park segregated into a distinct branch separate from the other more urban populations. CONCLUSION: Environmental heterogeneity may be affecting the wing shape variation of Cx. nigripalpus populations.

Wing geometry of Triatoma sordida (Hemiptera: Reduviidae) populations from Brazil.

Triatoma sordida has a widespread distribution in Argentina, Bolivia, Brazil, Paraguay, and Uruguay and is frequently found in peridomestic environments. We investigated size and shape variability of T. sordida wings across Brazil. Field-collected adults from twelve populations were studied. For each individual female, seven landmarks on the right wing were digitalized. Shape variables derived from Procrustes superimposition were used in Principal Component Analysis (PCA). Wing size and shape variations among populations was explored by means of ANOVA. Wing centroid size was significantly different among T. sordida populations; specimens from Bahia (East) were larger than those of Mato Grosso do Sul (West). PCA based on wing shape variables showed low wing shape variability. These results reinforce previous data showing low genetic variability among T. sordida populations from Brazil.

Composition and diversity of mosquitoes (Diptera: Culicidae) in urban parks in the South region of the city of São Paulo, Brazil / Composição e diversidade de mosquitos (Diptera: Culicidae) em parques urbanos localizados na região Sul do município de São Paulo, Brasil

Abstract Many parks in the city of São Paulo contain remnants of Atlantic Forest. Of the 30 municipal parks in the South of the city, we investigated two in this study (Santo Dias Park and Shangrilá Park) in order to survey their mosquito fauna and investigate the presence of potential bioindicators of environmental conditions and vectors of human pathogens. Mosquitoes were collected monthly between March 2011 and February 2012 using aspirators, Shannon and CDC traps for adult mosquitoes and larval dippers and suction samplers for immature forms. Sampling effort was evaluated by plotting a species accumulation curve, and total richness was estimated using the first-order jackknife. To compare the diversity between the two parks Shannon and Simpson diversity indexes were calculated. Species similarity was compared by the Sorensen similarity index. In all, 8,850 specimens were sampled in both parks. Collections in Santo Dias Park yielded 1,577 adult mosquitoes and 658 immature individuals distributed in seven genera (Aedes, Anopheles, Culex, Limatus, Mansonia, Toxorhynchites and Wyeomyia) and 27 taxonomic units. Among the adult mosquitoes collected, Culex nigripalpus .and Aedes fluviatilis were the most abundant, while the most abundant immature forms were Cx. imitator, Wy. davisi, Wy. galvaoi and Ae. albopictus. Collections in Shangrilá Park yielded 4,952 adult specimens and 1,663 immature forms distributed in eight genera (Aedes, Anopheles, Culex, Limatus, Mansonia, Toxorhynchites, Uranotaenia and Wyeomyia) and 36 taxonomic units. Species accumulation curves in both parks were close to the asymptote, and the total richness estimate was close to the observed richness. Although the observed species richness was higher in the Shangrilá Park, there was no statistically significant difference between the diversity indexes measured. Regarding species composition, the two sites shared 16 species, including those of epidemiological importance such as Culex nigripalpus, Cx. quinquefasciatus, Aedes albopictus and Ae. aegypti. As some of the mosquito taxa found are bioindicators of environmental conditions and have epidemiological potential to carry pathogens, we recommend that urban parks should be included in official mosquito surveillance programs, and regular surveys carried out to detect circulating arboviruses.

Resumo Parques urbanos do município de São Paulo contêm remanescentes de Mata Atlântica. No sul da cidade há 30 parques municipais, sendo os parques Santo Dias e Shangrila alvos deste estudo. Este estudo teve a proposta de levantamento da fauna de culicídeos desses dois parques no sul da cidade de São Paulo e avaliar a presença de potenciais bioindicadores e espécies vetoras de patógenos aos seres humanos. Os mosquitos foram coletados mensalmente entre março de 2011 e fevereiro de 2012, com aspiradores, armadilhas de Shannon e CDCs para mosquitos adultos e conchas entomológicas e bombas manuais de sucção para os imaturos. O esforço amostral foi avaliado por traçar uma curva de acumulação de espécies, e a riqueza total foi estimada pelo método jackknife de primeira ordem. Para comparar a diversidade entre os dois parques, foram calculados os índices de diversidade de Shannon e de Simpson. A similaridade na composição de espécies foi comparada pelo índice de similaridade de Sorensen. Foram coletados um total de 8.850 espécimes de culicídeos em ambos os parques. Coletas no parque Santo Dias renderam 1.577 mosquitos adultos e 658 imaturos, distribuídos em sete gêneros (Aedes, Anopheles, Culex, Limatus, Mansonia, Toxorhynchites e Wyeomyia) e 27 unidades taxonômicas: Culex nigripalpus, e Aedes fluviatilis foram as mais abundantes unidades taxonômicas coletadas como adultos, enquanto em formas imaturas, as espécies mais abundantes coletadas foram Cx. imitator, Wy. davisi, Wy. galvaoi e Ae. albopictus. Coletas no parque Shangrilá renderam 4.952 espécimes como adultos e 1.663 formas imaturas, distribuídas em oito gêneros (Aedes, Anopheles, Culex, Limatus, Mansonia, Toxorhynchites, Uranotaenia e Wyeomyia) e 36 unidades taxonômicas.. As curvas de acúmulo de espécies em ambos os parques ficaram perto da assíntota, e as estimativas de riqueza total foram próximas às riquezas observadas. Apesar da riqueza observada ter sido maior no parque Shangrilá, não houve diferença estatisticamente significante entre os índices de diversidade mensurados. Em relação à composição de espécies os dois locais compartilharam 16 espécies, incluindo as de maior importância epidemiológica como Culex nigripalpus, Cx. quinquefasciatus, Aedes albopictus e Ae. aegypti. Alguns táxons de culicídeos são bioindicadores de condições ambientais nas áreas ou possuem potencial para veicular patógenos. Atenção deve ser dada a parques urbanos, com inclusão destes locais nos programas oficiais de vigilância entomológica e investigações periódicas na circulação de arbovírus.

Description of Rhodnius marabaensis sp. n. (Hemiptera, Reduviidae, Triatominae) from Pará State, Brazil.

Rhodnius marabaensissp. n. was collected on 12 May 2014 in the Murumurú Environmental Reserve in the city of Marabá, Pará State, Brazil. This study was based on previous consultation of morphological descriptions of 19 Rhodnius species and compared to the identification key for the genus Rhodnius. The examination included specimens from 18 Rhodnius species held in the Brazilian National and International Triatomine Taxonomy Reference Laboratory in the Oswaldo Cruz Institute in Rio de Janeiro, Brazil. The morphological characteristics of the head, thorax, abdomen, genitalia, and eggs have been determined. Rhodnius prolixus and Rhodnius robustus were examined in more detail because the BLAST analysis of a cyt-b sequence shows they are closely related to the new species, which also occurs in the northern region of Brazil. The most notable morphological features that distinguish Rhodnius marabaensissp. n. are the keel-shaped apex of the head, the length of the second segment of the antennae, the shapes of the prosternum, mesosternum and metasternum, the set of spots on the abdomen, the male genitalia, the posterior and ventral surfaces of the external female genitalia, and the morphological characteristics of the eggs. Rhodnius jacundaensis Serra, Serra & Von Atzingen (1980) nomen nudum specimens deposited at the Maraba Cultural Center Foundation - MCCF were examined and considered as a synonym of Rhodnius marabaensissp. n.

Mitochondrial PCR-RFLP Assay to Distinguish Triatoma brasiliensis macromelasoma from Triatoma brasiliensis brasiliensis Subspecies (Hemiptera: Reduviidae).

Triatoma brasiliensis sensu lato (s.l.), the main vector of Chagas disease in northeastern Brazil, is a species complex comprising four species, one with two subspecies (T. brasiliensis brasiliensis, T. brasiliensis macromelasoma, T. juazeirensis, T. sherlocki, and T. melanica), and each taxon displaying distinct ecological requirements. In order to evaluate the genetic relationships among nine T. brasiliensis s.l. populations from northeastern Brazil, we analyzed their mitochondrial cytochrome c oxidase subunit 1 sequences and suggested a PCR-RFLP assay to distinguish between T. b. macromelasoma and T. b. brasiliensis subspecies. All the specimens were morphologically identified as T. b. brasiliensis. The resulting phylogenies identified two major clades that are congruent with the geographical populations studied. Based on collection sites and in accordance with type-location, one clade was identified as the subspecies T. b. macromelasoma. The second clade grouped T. b. brasiliensis populations. Restriction endonuclease sites were observed in the sequences and used in PCR-RFLP assays, producing distinct fingerprints for T. b. macromelasoma and T. b. brasiliensis populations. The results suggest that these are different species and that gene flow occurs only among T. b. brasiliensis populations, possibly associated with human activity in the area.

Análise das relações taxonômicas e sistemáticas entre espécies de triatomíneos (Hemiptera, Reduviidae) de colônias mantidas pelo Serviço Especial de Saúde de Araraquara, inferida de seqüências do 16S rDNA mitocondrial / Taxonomic and systematic analysis among triatomine bug species (Hemiptera, Reduviidae) from colonies of the Special Health Service of Araraquara (SESA), inferred from 16S rDNA mitocondrial sequences

Nucleotide sequences of mitocondrial 16S rDNA gene were analyzed in 14 populations of triatomine bugs from colonies maintained by SESA, Araraquara, São Paulo, Brazil, comparing them with sequences of same gene found in the GenBank. The fragments varied from 311 to 317 bp, with low intra-specific genetic distance (0 percent to 0.6 percent), with exception of the relationship among Triatoma sordida specimens (1 percent) and T. brasiliensis specimens (1.3 percent) that were attributed to different geographical populations. The paraphily of Rhodniini and Panstrongylus genus was evidenced by this analyses, confirming previous results between these and the narrow relationships of R. prolixus with R. robustus and of T. infestans with T. platensis. The relationship between T. maculata and T. pseudomaculata has not been resolved, since these taxa appeared both in monophyly and as in paraphyly: T. pseudomaculata (SESA) is grouped with T. maculata (sequence of GenBank) and the associated T. brasiliensis (SESA), while T. maculata (SESA) appears grouped with T. pseudomaculata of SESA and GenBank. The results evidence the utility of the gene 16S as molecular marker in the species of triatomine bugs and its importance in systematic and taxonomy questions. There is a necessity of new studies including new molecular markers associated with classic systematic characters of morphology, ecology and behavior to adjust systematic decisions since those will be not of only systematic impact but, for new strategies of control.

Foram analisadas seqüências de nucleotídeos do gene 16S do rDNA mitocondrial em 14 populações de triatomíneos mantidos em colônias no insetário SESA de Araraquara- SP, comparando-as com seqüências do mesmo gene disponíveis no GenBank. Os fragmentos variaram de 311 a 317 pb com baixa variação intra-específica entre as distâncias genéticas (0 por cento a 0,6 por cento), exceto para os relacionamentos entre espécimes de Triatoma sordida (1 por cento) e espécimes de T. brasiliensis (1,3 por cento) atribuídos a populações geográficas diferentes. A parafilia de Rhodniini e do gênero Panstrongylus foi evidenciada pelas analises, confirmando resultados anteriores entre estes e os estreitos relacionamentos de R. prolixus com R. robustus e de T. infestans e T. platensis. O relacionamento entre T. maculata e T. pseudomaculata não foi solucionado, uma vez que, esses táxons apareceram tanto em monofilia quanto em parafilia: T. pseudomaculata (SESA) está agrupado com T. maculata (seqüência do GenBank) e associados a T . brasiliensis (SESA), enquanto T. maculata (SESA) aparece agrupado com T. pseudomaculata do SESA e do GenBank. Os resultados evidenciam a utilidade do gene 16S como marcador de espécies de triatomíneos e sua importância em questões de sistemática e taxonomia. Há necessidade de novos estudos envolvendo outros marcadores associados a caracteres sistemáticos clássicos de morfologia, ecologia e comportamento para decisões sistemáticas adequadas uma vez, que teriam impacto não apenas sistemático mas, para as estratégias de controle.