Biology of the first generation of a laboratory colony of Nyssomyia intermedia (Lutz & Neiva, 1912) and Nyssomyia neivai (Pinto, 1926) (Diptera: Psychodidae)

The phlebotomine sand flies Nyssomyia intermedia (Lutz & Neiva, 1912) and Nyssomyia neivai (Pinto, 1926) are very close and may be involved in the transmission of Leishmania spp. Ross, 1903 in Brazil. The biology of the first laboratory-reared generations of these species, descended from insects captured in Além Paraíba (N. intermedia) and Corinto (N. neivai) in the Brazilian state of Minas Gerais, is described here. The captured females were fed on hamsters and maintained individually in rearing pots. Laboratory temperature and relative humidity were maintained at 25-26 masculineC and 80% respectively. The productivity of the first generation of N. intermedia was greater than that of N. neivai, and its development time clearly shorter, particularly for the second and third larval instars.     

Nyssomyia intermedia (Lutz & Neiva, 1912) and Nyssomyia neivai (Pinto, 1926) (Diptera: Psychodidae: Phlebotominae) geographical distribution and epidemiological importance

Nyssomyia intermedia (Lutz & Neiva 1912) and N. neivai (Pinto 1926) are possible vectors of tegumentary leishmaniasis in some regions of Brazil. Further, the latter was until recently, considered a junior synonym of the former. This study has the purpose of updating our knowledge of the geographical distribution of these species, based on specimens deposited at the collection of the Centro de Pesquisas René Rachou-Fiocruz, Faculdade de Saúde Pública-Universidade de S?o Paulo, and on data presented by literature as also to associate this distribution with the cutaneous leishmaniasis cases reported. It has been reported that N. intermedia occurs in the states of the Northeastern Region, in Rio de Janeiro, Espírito Santo, on the northern coast of S?o Paulo, in eastern Minas Gerais, Mato Grosso do Sul, and Goiás, close to the border with Minas Gerais and Bahia. N. neivai occurs in the Southern Region, southern coast and in western S?o Paulo, southern and western Minas Gerais, southern Goiás, and southern Pará, beyond Argentina, Bolivia, and Paraguay. It is important to highlight that N. intermedia and N. neivai occur in sympatry in Minas Gerais and S?o Paulo. N. intermedia or N. neivai are predominant or are captured abundantly in several cutaneous leishmaniasis foci in the Southeastern and Southern regions of Brazil.     

Variations in the gonostyle of Nyssomyia intermedia (Lutz & Neiva) (Diptera: Psychodidae)

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.     

Preliminary study on feeding preference of Nyssomyia neivai (Pinto) and Nyssomyia whitmani (Antunes & Coutinho) (Diptera: Psychodidae) in a rural area of the State of Paraná, South Brazil

Due to the importance of sandflies in the American cutaneous leishmaniasis epidemiology, we investigated the feeding preference of sandflies related to domestic animals common in domiciliary environments in endemic rural areas, where this disease occurs. The sandflies were collected with Falc?o light traps, installed in cages, each one containing one domestic animal kind (pig, dog, rabbit or chicken), in Recanto Marista, in Doutor Camargo municipality. Forty five collections were made, totaling 135h of collections per trap. A total of 2,854 sandflies of the following species were collected: Brumptomyia brumpti (Larousse), Evandromyia correalimai (Martins, Coutinho & Luz), Evandromyia cortelezzii (Brèthes), Expapillata firmatoi (Barreto, Martins & Pellegrino), Micropygomyia ferreirana (Barreto, Martins & Pellegrino), Migonemyia migonei (Fran?a), Nyssomyia neivai (Pinto), Nyssomyia whitmani (Antunes & Coutinho), Pintomyia fischeri (Pinto), Pintomyia monticola (Costa Lima), Pintomyia pessoai (Coutinho & Barreto), Psathyromyia lanei (Barreto & Coutinho) and Psathyromyia shannoni (Dyar). The dominant species was N. neivai. N. neivai and N. whitmani did not show any feeding preference towards the animals used as bait. It was concluded that N. neivai and N. whitmani are opportunist and, probably, the females adjust their feeding habits to the availability of hosts, suggesting their feeding eclecticism in the anthropic environments.     

Correlation of male genital filaments and female spermathecal ducts in New World sand flies of the Lutzomyia intermedia species complex (Diptera: Psychodidae,Phlebotominae)

The lengths of the male genital filaments and female spermathecal ducts were measured in phlebotomine sand flies of the Lutzomyia intermedia species complex and the ratios between these characters calculated. Ratios for L. intermedia s. s. from Northeast vs Southeast Brazil (Espírito Santo and Minas Gerais), Espírito Santo/Minas Gerais vs Rio de Janeiro/S?o Paulo and L. intermedia vs L. neivai were significantly different at P < 0.1, 0.05 and 0.01 respectively when compared using ANOVA. The spermathecal ducts and genital filaments of L. intermedia were significantly longer than those of L. neivai (P < 0.01) and could be used to differentiate these species. The taxonomic and biological significance of these differences is discussed.     

Distinction of males of the Lutzomyia intermedia (Lutz & Neiva, 1912) species complex by ratios between dimensions and by an artificial neural network (Diptera: Psychodidae, Phlebotominae)

The females of the two species of the Lutzomyia intermedia complex can be easily distinguished, but the males of each species are quite similar. The ratios between the extra-genital and the genital structures of L. neivai are larger than those of L. intermedia s. s., according to ANOVA. An artificial neural network was trained with a set of 300 examples, randomly taken from a sample of 358 individuals. The input vectors consisted of several ratios between some structures of each insect. The model was tested on the remaining 58 insects, 56 of which (96.6%) were correctly identified. This ratio of success can be considered remarkable if one takes into account the difficulty of attaining comparable results using traditional statistical techniques.     

Micro-geographical variation among male populations of the sandfly, Lutzomyia (Nyssomyia) intermedia, from an endemic area of American cutaneous leishmaniasis in the state of Rio de Janeiro, Brazil

The genetic relationships among male Lutzomyia (Nyssomyia) intermedia (Lutz & Neiva) (Diptera: Psychodidae) from three populations from the same endemic area of American cutaneous leishmaniasis (ACL) in the state of Rio de Janeiro, Brazil, were compared. The sandflies were collected in three ecologically different habitats: domestic, extra-domestic and sylvatic over a total range of 800 m. Three molecular markers were employed to assess population variation. Based on MLEE markers, it could not be concluded that the three populations do not belong to the same gene pool (F(st) = 0.005). No within-population departure from Hardy-Weinberg equilibrium was detected (P < 0.05) and they presented the same level of gene variation. The number of migrants (Nm) indicated that at least 50 individuals per generation migrated between the three habitats. RAPD-PCR markers revealed that, except for the primer five, all were polymorphic. Phenetic analysis of the genotypes showed the presence of two principal clusters corresponding to: (1) domestic plus extra-domestic and (2) sylvatic. Unique genotypes were observed in each population. The sylvatic population was the most polymorphic, showing the largest number of genotypes and low level of similarity between them. Three mtDNA gene markers were studied by SSCP analysis. The most frequent haplotype for each marker ranged in frequency from 60 to 87% and individuals with unique haplotypes varied from 1 to 5%. Interestingly, the SSCP analysis showed a low level of polymorphism within populations. The disagreement between the different molecular markers observed and the hypothesis that L. intermedia could be participating in the transmission cycle of Leishmania (Viannia) braziliensis in environments ranging from the interior of human dwellings to the forest, are discussed.     

Lutzomyia (Nyssomyia) whitmani s.l . (Antunes & Coutinho, 1939)(Diptera: Psychodidae: Phlebotominae): geographical distribution and the epidemiology of American cutaneous leishmaniasis in Brazil--mini-review

The aim of the present study, in view of the widespread geographical distribution of Lutzomyia (Nyssomyia) whitmani s.l. in Brazil, in close association with the regions of transmission of Leishmania (Viannia) braziliensis and L. (V.) shawi, is to investigate the distribution of this sand fly species and American cutaneous leishmaniasis (ACL) in relationship to vegetation and landscape ecology throughout Brazil. Thematic maps were elaborated by the MapInfo programme, giving information on the spatial distribution of L. whitmani s.l., in accordance with types of vegetation and foci of ACL. With regards to the known areas of transmission of ACL in Brazil, it is notable that L. (N.) whitmani s.l. occurs in most of them, where it has been implicated as a possible vector of L. (V.) braziliensis. The presence of L. whitmani s.l. has been registered in 26 states, the one exception being Santa Catarina; in some states such as Roraima, Acre, Tocantins, and Mato Grosso do Sul this sand fly species has been recorded in a large number of municipalities. L. whitmani s.l. has been found in association with a variety of vegetation types, including the Amazonian forest, savanna ("campos cerrados"), and northeastern savanna ("caatingas nordestinas" or "savana estépica").     

Larval habitats of sand flies in rural areas of southern Brazil

We report the results of an investigation of natural larval sand fly habitats in the Recanto Marista, Doutor Camargo municipality, Paraná state, Brazil, from May, 2010 to August, 2012. We used Alencar emergence traps (AT), experimental traps (ET), and soil samples incubated in a biochemical oxygen demand incubator. Eight sand flies were collected with ATs. One specimen was collected with an ET and 21 were collected in soil samples. The collected species were Brumptomyia brumpti, Micropygomyia ferreirana, Migonemyia bursiformis, Migonemyia migonei, Nyssomyia neivai, Nyssomyia whitmani, and Pintomyia pessoai. The laval habitats of sand flies were located in the Recanto Marista, especially between tree roots, but the number of adults that emerged in the traps and soil samples was small despite the high density of sand flies that has been recorded in the Recanto Marista.     

Diversity, distribution and abundance of sandflies (Diptera: Psychodidae) in Parana State, Southern Brazil

Due to the spread of Visceral leishmaniasis in Brazilian territory, this research was undertaken with the objective to identify the sandfly fauna and aspects of the species behavior in forest and anthropic environments in Paraná State, Brazil. The sandfly captures were sporadically carried out in 37 municipalities of the Paraná state, during the period from March 2004 to November 2005. Each municipality was once visited, when Falc?o traps were installed during three consecutive nights in forested fragments and anthropic environments; captures with Shannon traps installed in forests and edge of forests, inspection of walls and electric aspiration in domiciles and forested areas were sporadically undertaken. The data analysis was based on estimates of frequency and abundance of the species, by five native landscapes. A total of 38,662 specimens belonging to 23 species were captured. Nyssomyia neivai (Pinto) (75.6%), Ny. whitmani (Antunes & Coutinho) (10.1%), Migonemyia migonei (Fran?a) (7.8%), Expapillata firmatoi (Barreto et al.) (2.1%) and Pintomyia fischeri (Pinto) (1.6%) together constituted 97.2% of the phlebotomine collected. High frequencies of sand flies were found in forests, henhouses, pig-pens and domiciles. The three most frequent species have been implicated in transmission of tegumentary leishmaniasis in Paraná and in the Brazilian Southeastern and Southern Regions. Lutzomyia longipalpis (Lutz & Neiva), the visceral leishmaniasis vector, was not found, however Lutzomyia gaminariai (Cordero et al.), female of which is similar to that of Lu. longipalpis was captured, raising the need to investigate its behavior, including as regard to the vectorial competence.     

Lutzomyia sand flies (Diptera: Psychodidae) from middle and lower Putumayo Department, Colombia, with new records to the country

A total of 4,840 phlebotomine sand flies from 54 localities in Putumayo department (=state), in the Colombian Amazon region, were collected in Shannon traps, CDC light traps, resting places and from human baits. At least 42 Lutzomyia species were registered for the first time to the department. Psychodopygus and Nyssomyia were the subgenera with the greatest number of taxa, the most common species being L. (N.) yuilli and L. (N.) pajoti. They were sympatric in a wide zone of Putumayo, indicating that they should be treated as full species (new status). Among the anthropophilic sand flies, L. gomezi and L. yuilli were found in intradomiciliar, peridomestic, urban or forest habitats. L. richardwardi, L. claustrei, L. nocticola and L. micropyga are reported for the first time in the Colombian Amazon basin. L. pajoti, L. sipani and L. yucumensis are new records for Colombia.     

External morphology of sensory structures of fourth instar larvae of neotropical species of phlebotomine sand flies (Diptera: Psychodidae) under scanning electron microscopy.

In the present study, some morphological structures of antennae, maxillary palps and caudal setae of fourth instar larvae of laboratory-reared phlebotomine sand flies (Lutzomyia longipalpis, L. migonei, L. evandroi, L. lenti, L. sericea, L. whitmani and L. intermedia) of the State of Ceará, Brazil, were examined under scanning electron microscopy. The antennal structures exhibited considerable variation in the morphology and position. A prominent digitiform distal segment has been observed only on the antenna of species of the subgenus Nyssomyia. The taxonomic relevance of this and other antennal structure is discussed. The papiliform structures found in the maxillae and the porous structures of the caudal setae of all species examined may have chemosensory function. Further studies with transmission electron microscopy are needed to better understand the physiological function of these external structures.     

Comparative anatomy of the cheek muscles within the Centromochlinae subfamily (Ostariophysi, Siluriformes, Auchenipteridae)

Glanidium melanopterum Miranda Ribeiro, a typical representative of the subfamily Centromochlinae (Siluriformes: Auchenipteridae), is herein described myologically and compared to other representative species within the group, Glanidium ribeiroi, G. leopardum, Tatia neivai, T. intermedia, T. creutzbergi, Centromochlus heckelii, and C. existimatus. The structure of seven pairs of striated cephalic muscles was compared anatomically: adductor mandibulae, levator arcus palatini, dilatator operculi, adductor arcus palatini, extensor tentaculi, retractor tentaculi, and levator operculi. We observed broad adductor mandibulae muscles in both Glanidium and Tatia, catfishes with depressed heads and smaller eyes. Similarities between muscles were observed: the presence of a large aponeurotic insertion for the levator arcus palatini muscle; an adductor arcus palatini muscle whose origin spread over the orbitosphenoid, pterosphenoid, and parasphenoid; and the extensor tentaculi muscle broadly attached to the autopalatine. There is no retractor tentaculi muscle in either the Glanidium or Tatia species. On the other hand, in Centromochlus, with forms having large eyes and the tallest head, the adductor mandibulae muscles are slim; there is a thin aponeurotic or muscular insertion for the levator arcus palatini muscle; the adductor arcus palatini muscle originates from a single osseous process, forming a keel on the parasphenoid; the extensor tentaculi muscle is loosely attached to the autopalatine, permitting exclusive rotating and sliding movements between this bone and the maxillary. The retractor tentaculi muscle is connected to the maxilla through a single tendon, so that both extensor and retractor tentaculi muscles contribute to a wide array of movements of the maxillary barbels. A discussion on the differences in autopalatine-maxillary movements among the analyzed groups is given.     

Molecular characterization of mutations of nlt-4, the pathway-specific regulatory gene which controls nitrate assimilation in Neurospora crassa

The nit-4 genes of three conventional Neurospora crassa mutations and of the closely related species, Neurospora intermedia, have been isolated by amplifying the genomic DNA with the polymerase chain reaction. Nucleotide sequencing has revealed that the three nit-4 mutants, alleles 15, 1214, and 2994, are the result of a missense mutation, a nonsense mutation and a frameshift mutation, respectively. The nucleotide sequence of the NIT4 protein coding region of a nit-4 mutant (allele 2994) and of N. intermedia have been determined and compared with that of wild-type N. crassa. The molecular characteristics confirm that the mutated gene of 2994 originated from N. intermedia and was introgressed into N. crassa. The polyglutamine domains of the N. crassa wild type, the 2994 mutant, or N. intermedia cannot replace an upstream glutamine-rich domain which is essential for nit-4 function.     

Ecological Niche Modelling Predicts Southward Expansion of Lutzomyia (Nyssomyia) flaviscutellata (Diptera: Psychodidae: Phlebotominae), Vector of Leishmania (Leishmania) amazonensis in South America,under Climate Change

Vector borne diseases are susceptible to climate change because distributions and densities of many vectors are climate driven. The Amazon region is endemic for cutaneous leishmaniasis and is predicted to be severely impacted by climate change. Recent records suggest that the distributions of Lutzomyia (Nyssomyia) flaviscutellata and the parasite it transmits, Leishmania (Leishmania) amazonensis, are expanding southward, possibly due to climate change, and sometimes associated with new human infection cases. We define the vector’s climatic niche and explore future projections under climate change scenarios. Vector occurrence records were compiled from the literature, museum collections and Brazilian Health Departments. Six bioclimatic variables were used as predictors in six ecological niche model algorithms (BIOCLIM, DOMAIN, MaxEnt, GARP, logistic regression and Random Forest). Projections for 2050 used 17 general circulation models in two greenhouse gas representative concentration pathways: “stabilization” and “high increase”. Ensemble models and consensus maps were produced by overlapping binary predictions. Final model outputs showed good performance and significance. The use of species absence data substantially improved model performance. Currently, L. flaviscutellata is widely distributed in the Amazon region, with records in the Atlantic Forest and savannah regions of Central Brazil. Future projections indicate expansion of the climatically suitable area for the vector in both scenarios, towards higher latitudes and elevations. L. flaviscutellata is likely to find increasingly suitable conditions for its expansion into areas where human population size and density are much larger than they are in its current locations. If environmental conditions change as predicted, the range of the vector is likely to expand to southeastern and central-southern Brazil, eastern Paraguay and further into the Amazonian areas of Bolivia, Peru, Ecuador, Colombia and Venezuela. These areas will only become endemic for L. amazonensis, however, if they have competent reservoir hosts and transmission dynamics matching those in the Amazon region.     

Description of Dendrorchis retrobiloba n. sp. (Digenea: Gorgoderidae) from Astyanax fasciatus (Teleostei: Characidae) from Southern Uruguay, with an emended diagnosis of the genus Dendrorchis

To date, the monospecific Dendrorchis is represented by D. neivai, reported from Brazil and parasitizing Brycon lundi. Dendrorchis retrobiloba n. sp. is described from the swim bladder of Astyanax fasciatus from Ca?ada del Dragón stream in Montevideo, southern Uruguay (34 degrees 47'S, 56 degrees 14'W), and the emended diagnosis of Dendrorchis is given. The diagnosis of Dendrorchis has been modified to include the following characters: Phyllodistominae with flattened body, wider in the region posterior to acetabulum. Intestinal ceca sinuous or not. Testes branched or with superficial lobes. Ovary with deep or superficial lobes. Seminal receptacle present or absent. Vitellaria ellongate or pyriform. Uterus extends to posterior region and to portion of body anterior to acetabulum. Eggs oval, variable size, thin-shelled, without opercula. Dendrorchis retrobiloba n. sp. differs from D. neivai by having pyriform vitellaria, ovary larger than the testes and not deeply lobed, wider distribution of the uterus, smaller egg size, testes with superficial lobes, oral sucker wider than the acetabulum, shorter esophagus, 2 terminal lobes at the posterior end of the body, and much smaller body length. Despite being found in different hosts, both species of Dendrorchis parasitize the swim bladder of freshwater Characidae.     

Catalogue of the type material of Phlebotominae (Diptera,Psychodidae) deposited in the Instituto Evandro Chagas,Brazil

The available type material of Phlebotominae (Diptera, Psychodidae) deposited in the “Coleção de Flebotomíneos” of the Instituto Evandro Chagas (ColFleb IEC) is now presented in an annotated catalogue comprising a total of 121 type specimens belonging to 12 species as follow: Nyssomyia richardwardi (2 female paratypes), Nyssomyia shawi (9 male and 25 female paratypes), Nyssomyia umbratilis (female holotype and 1 female paratype), Nyssomyia yuilli yuilli (1 male and 1 female paratypes), Pintomyia gruta (1 male and 2 female paratypes), Psychodopygus lainsoni (2 male syntypes), Psychodopygus leonidasdeanei (male holotype, female “allotype” and 45 female paratypes), Psychodopygus llanosmartinsi (2 female paratypes), Psychodopygus wellcomei (1 male and 4 female “syntypes”), Trichophoromyia readyi (male holotype, female “allotype” and 1 male paratype), Trichophoromyia adelsonsouzai (male holotype, 13 male 5 female paratypes), and Trichophoromyia brachipyga (1 male paratype).     

Co‐occurrence and seasonal and environmental distributions of the sandflies Lutzomyia longipalpis and Nyssomyia whitmani in the city of Puerto Iguazú, northeastern Argentina

The aim of this work was to study the distribution of Phlebotominae (Diptera: Psycodidade) abundance in time and space in an area in northeastern Argentina with vector transmission of visceral and tegumentary leishmaniasis. For this, 51 households were selected using a ‘worst scenario’ criterion where one light trap was set during two consecutive nights in peridomiciles in the transitions between the four seasons, and the environment was surveyed simultaneously. The relationships of phlebotomine assemblage structure and the most abundant species with seasonality and environmental variables were evaluated using a canonical correspondence analysis and generalized linear mixed models, respectively. A total of 5110 individuals were captured. Lutzomyia longipalpis (Lutz & Neiva, 1912) and Nyssomyia whitmani (Antunes & Coutinho, 1939) were the most abundant species captured in all samplings (98.3% of the total capture). The period of highest abundance of Lu. longipalpis was early autumn, and it was distributed in the most urbanized areas. Nyssomyia whitmani occupied mainly the less urbanized areas, showing peaks of abundance in early spring and summer. Other species were captured in low numbers and showed seasonal?spatial variations similar to those of Ny. whitmani . We confirmed Leishmania spp. vector persistence throughout the year in spatial patches of high abundance even during the less favorable season.     

Dispersal pattern of the sand fly Lutzomyia neivai (Diptera: Psychodidae) in a cutaneous leishmaniasis endemic rural area in Southeastern Brazil

The dispersal pattern of the sand fly Lutzomyia neivai was studied through mark-release-recapture experiments in an American cutaneous leishmaniasis endemic rural area in Southeastern Brazil. Over 6500 specimens were marked with fluorescent powder and released in forest edge and peridomicile habitats from August to November 1999, February and April 2000. Recapture attempts were made using Shannon and CDC traps up to eight successive nights after releases. A total of 493 (7.58%) specimens were recaptured. The number of recaptured males and females of L. neivai in CDC traps was not affected by the distance between the trap and the release points. Approximately 90% of males and females recaptured in CDC traps were caught up to 70 m from the release points. The maximum female flight range recorded was 128 m. The average flight range per day was less than 60 m for males and females. Of the flies released in forest edge, approximately 16% of the recaptured females were caught in Shannon traps in the peridomicile habitat. The results indicate that the movements of L. neivai are spatially focal and the possibility of dispersion from forest to peridomicile habitat may be an important way of contracting leishmaniasis in dwellings.     

Resistance to starvation of Rhodnius neivai Lent, 1953 (Hemiptera: Reduviidae: Triatominae) under experimental conditions

The period of resistance to starvation and the loss of weight until death of Rhodnius neivai in all stages of development were studied. Work was based on experiments conducted under controlled laboratory conditions. One hundred specimens of each nymphal instar were observed: 50 were fed on chicken and 50 on rabbit. Adult females and males were kept together and fed on each host. All bugs were weighed weekly until death. Laid eggs were collected weekly and observed during five weeks to obtain hatchability. Resistance to starvation was similar with both hosts and increased with the evolutionary stage, excepting the 5th nymphal instar and adults. With both hosts, loss of weight was abrupt in the first week and steady in the following weeks. In adults, on the first weeks after eating, there was little or no mortality, after which mortality increased rapidly with the starving time. Reproductive output was higher in the bugs fed on rabbit. R. neivai is among the least resistant triatomine species.