Stomodaeal valve ultrastructure in the sandfly Lutzomyia longipalpis (Diptera: Psychodidae)

The stomodaeal valve and adjacent cardia region of the alimentary canal of the phlebotomine sandfly Lutzomyia longipalpis have been studied using electron microscopy. With the scanning electron microscope (SEM), numerous pit-like structures of different sizes were found in the surface of the valve of male and female flies. Under the transmission electron microscope (TEM), the cells adjacent to the pores were shown to have numerous mitochondria and to lack neural innervation. It is suggested that the pit-like structures are probably secretory. A typical basiconic sensilla was found on the inner side of the oesophagus at the junction with the stomodaeal valve, with at least one dendrite which extended to the tip, indicating a chemosensory role. It is likely that this type of sensilla may control the movement of the stomodaeal valve after contact with fluids entering the mid-gut and may be responsible for diverting the sugar meal to the crop by closure of the valve.     

A revised numerical chaetotaxy for neotropical Phlebotomine sandfly larvae (Diptera: Psychodidae)

Phlebotomine larval taxonomy is briefly reviewed with particular reference to New World species, of which under one fifth have been adequately described as immature stages. A new numerical chaetotaxy is proposed following studies on the larvae of six species from Brazil. Setal numeration is used in a manner which demonstrates apparent segmental homologies and the use of letter designations for some setae has been reduced. With two exceptions the mesothoracic, metathoracic and abdominal setae are homologized with those of the posterior prothorax, and the anterior prothoracic setae are regarded as atypical.     

Geographic distribution of phlebotomine sandfly species (Diptera: Psychodidae) in Central-West Brazil

This study updates the geographic distributions of phlebotomine species in Central-West Brazil and analyses the climatic factors associated with their occurrence. The data were obtained from the entomology services of the state departments of health in Central-West Brazil, scientific collections and a literature review of articles from 1962-2014. Ecological niche models were produced for sandfly species with more than 20 occurrences using the Maxent algorithm and eight climate variables. In all, 2,803 phlebotomine records for 127 species were analysed. Nyssomyia whitmani, Evandromyia lenti and Lutzomyia longipalpis were the species with the greatest number of records and were present in all the biomes in Central-West Brazil. The models, which were produced for 34 species, indicated that the Cerrado areas in the central and western regions of Central-West Brazil were climatically more suitable to sandflies. The variables with the greatest influence on the models were the temperature in the coldest months and the temperature seasonality. The results show that phlebotomine species in Central-West Brazil have different geographical distribution patterns and that climate conditions in essentially the entire region favour the occurrence of at least one Leishmania vector species, highlighting the need to maintain or intensify vector control and surveillance strategies.     

Sugar feeding and fluid destination control in the phlebotomine sandfly Lutzomyia longipalpis (Diptera: Psychodidae)

Sandfly feeding behaviour and destination of coloured sugar meals in the gut of Lutzomyia longipalpis were investigated with particular attention to the role of the crop. Sandflies were able to ingest sugar from liquid drops, microcapillaries, a slice of pear and even sugar powder. In most cases the flies adopted a 'sugar feeding mode' with raised palps. As the fruit dried, flies of both sexes fed by piercing the tissue with the proboscis. All sugar-fed flies had a full crop plus a small amount of sugary fluid in the thoracic mid-gut, i.e. past the stomodaeal valve. Dissections of flies interrupted during feeding showed that the very first trace of sugar passed through the stomodaeal valve, but that the rest of the meal was diverted into the crop. This suggests that closure of the stomodaeal valve is initiated only after a small volume of sugar solution has passed through it.     

Ultrastructure of male sex pheromone glands in abdominal tergites of five Lutzomyia sandfly species (Diptera: Psychodidae)

The fine structure of male sex pheromone disseminating structures on abdominal segments of five species of Lutzomyia sandflies (Diptera: Psychodidae) was analyzed. Scanning electron microscopy showed that in Lutzomyia cruzi [Mem. Inst. Oswaldo Cruz 33 (1938) 349] and Lutzomyia longipalpis [Mem. Inst. Oswaldo Cruz 4 (1912) 84], the disseminating structures are located in pale spots on abdominal tergites IV or III and IV and are morphologically similar, appearing as small round cuticular elevations with a central pore. Observation of abdominal tergites of L. longipalpis pupae showed that the spots, but not the structures, are already present in this stage. In Lutzomyia lenti [Mem. Inst. Oswaldo Cruz 33 (1938) 349] and Lutzomyia carmelinoi [Mem. Inst. Oswaldo Cruz 81 (1986) 323] adult males, the disseminating structures are present on tergal segments V and VI, where pale spots could not be observed, and appear as apple-like elevations with a central pore. In Lutzomyia renei, a single disseminating structure is found at the anterior region of tergal segment VI. Transmission electron microscopy was used to analyze the gland cell fine structure in L. cruzi. Several class III gland cells are located side by side in the fourth abdominal segment. Each epidermal secretory cell contains a small reservoir and a short outlet channel through the cuticle.     

Studies on sandfly fauna (Diptera: Psychodidae) in a foci of cutaneous leishmaniasis in mesquita, Rio de Janeiro State, Brazil

In a study on vectors of cutaneous leishmaniasis in Mesquita, Rio de Janeiro State, Brazil, eleven species of sandflies were found: Lutzomyia longipalpis, L. fischeri, L. quinquefer, L. intermedia, L. whitmani, L. shannoni, L. cortelezzii, L. edwardsi, L. migonei, L. lanei and L. firmatoi. Lutzomyia intermedia was the predominant species. It was collected indoors but in much higher numbers outdoors, in the close vicinity of houses. In comparative captures it was found to feed more frequently on equines, and less frequently on man and dogs. Studies, using man and animals as baits, showed that blood-feeding started in the evening, and remained stable during the night, until early morning. June, August and October were the months of highest density.     

Identification of oviposition attractants for the sandfly Lutzomyia longipalpis (Diptera: Psychodidae) in volatiles of faeces from vertebrates

Abstract. Extracts of volatiles from rabbit and chicken faeces preferentially attracted gravid sandflies, Lutzomyia longipalpis (Lutz and Neiva), in an oviposition bioassay. In electrophysiology experiments, the same extracts selectively stimulated two olfactory cells while inhibiting another in ascoid sensilla on the antennae of these flies. Analysis of faeces volatiles by gas chromatography linked to ascoid sensillum recording revealed two early eluting electrophysiologically active components of rabbit faeces. These active compounds were identified in both rabbit and chicken faeces volatile extracts by gas chromatography -mass spectrometry as hexanal and 2-methyl-2-butanol. Hexanal stimulated one cell type and inhibited another, whereas 2-methyl-2-butanol stimulated a third cell type. A 1:l mixture of synthetic hexanal and 2-methyl-2-butanol elicited the same targeted oviposition response from gravid females on the treatment septum of the bioassay as did the total volatile extract of rabbit or chicken faeces.
The monoterpenes α(+)-pinene (plus some optical and positional isomers) and a-terpinene activated a separate cell type, whereas benzaldehyde stimulated the same receptor as hexanal, but with a higher threshold. Furthermore, an olfactory cell selectively tuned to the perception of the male sex pheromone of this species was also found in the ascoid sensillum.     

Phylogenetic relationships in the subfamily Psychodinae (Diptera,Psychodidae)

Espíndola, A., Buerki, S., Jacquier, A., Je?ek, J. & Alvarez, N. (2012). Phylogenetic relationships in the subfamily Psychodinae (Diptera, Psychodidae). —Zoologica Scripta, 41, 489–498. Thanks to recent advances in molecular systematics, our knowledge of phylogenetic relationships within the order Diptera has dramatically improved. However, relationships at lower taxonomic levels remain poorly investigated in several neglected groups, such as the highly diversified moth‐fly subfamily Psychodinae (Lower Diptera), which occurs in numerous terrestrial ecosystems. In this study, we aimed to understand the phylogenetic relationships among 52 Palearctic taxa from all currently known Palearctic tribes and subtribes of this subfamily, based on mitochondrial DNA. Our results demonstrate that in light of the classical systematics of Psychodinae, none of the tribes sensu Je?ek or sensu Vaillant is monophyletic, whereas at least five of the 12 sampled genera were not monophyletic. The results presented in this study provide a valuable backbone for future work aiming at identifying morphological synapomorphies to propose a new tribal classification.     

The effect of male sandfly pheromone and host factors as attractants for female Lutzomyia longipalpis (Diptera: Psychodidae)

Abstract Male Lutzomyia longipalpis (Lutz & Neiva) pheromone extracted from tergal glands combined with heat, carbon dioxide gas and hamster urine was found to be attractive to virgin female sandflies. The host factors if offered in the absence of pheromone or a heat source were, however, unattractive to female flies. Pheromone stored for 6 days and then combined with the same host components remained attractive to female flies but storage failed to enhance its attractiveness. The effects of carbon dioxide gas, hamster urine, chicken uropygial gland extract and octenol plus acetone on the responses of females were inconsistent. A ceramic battery powered heat source was tested with pheromone and host attractants and showed potential for field application in developing a pheromone trap.     

The immature stages of some Phlebotomine sandflies from Brazil (Diptera: Psychodidae)

Immature stages of Lutzomyia anduzei (Rozeboom), L.antunesi (Coutinho), L. flaviscutellata (Mangabeira), Psychodopygus davisi (Root), Ps.paraensis (Costa Lima) and Ps.lainsoni Fraiha & Ward are described from specimens reared in the laboratory. The Lutzomyia species are members of the intermedia group, though L. flaviscutellata larvae more closely resemble those of Psychodopygus species. The larvae of L.anduzei and L. antunesi are separable on minor setal differences and are similar to previously described members of the intermedia group. Ps. paraensis is distinguishable from the other two Psychodopygus species described, by the absence of prothoracic seta 2. Head seta 5 of Ps.davisi is about twice the length of that in Ps. paraensis and larvae of Ps.lainsoni are pale and characterized by colourless body setae 4 and 10. A key to the fourth instar larvae is given which includes all the known larvae of the intermedia group and of the genus Psychodopygus. Morphological differences in the Psychodopygus larvae do not appear to reflect the separation of the adults into series.     

(1S,3S,7R)-3-methyl-alpha-himachalene from the male sandfly Lutzomyia longipalpis (Diptera: Psychodidae) induces neurophysiological responses and attracts both males and females

Lutzomyia longipalpis adult males form leks on or near hosts and release (1S,3S,7R)-3-methyl-alpha-himachalene from their tergal glands to lure females to the same site for mating and feeding. Here we have examined whether the male-produced attractant could also serve as a male aggregation stimulus. High resolution chiral capillary gas chromatography analysis of male tergal gland extracts, synthetic (1S,3S,7R)-3-methyl-alpha-himachalene, and a synthetic mixture of all isomers of 3-methyl-alpha-himachalene, was coupled to electrophysiological recordings from ascoid sensillum receptor cells in antennae of male and female sandflies. Receptor cells of both sexes responded only to the main component of the male tergal gland extract that eluted at the same retention time as (1S,3S,7R)-3-methyl-alpha-himachalene. Furthermore, of the eight 3-methyl-alpha-himachalene isomers in the synthetic mixture only the fraction containing (1S,3S,7R)-3-methyl-alpha-himachalene, co-eluting with an isomer of (1S*,3S*,7S*)-3-methyl-alpha-himachalene, elicited an electrophysiological response from male and female ascoid sensillum receptor cells. Both males and females flew upwind in a wind tunnel towards a filter paper disk treated with either 4-6 male equivalents of the tergal gland extract, pure (1S,3S,7R)-3-methyl-alpha-himachalene or the synthetic mixture of eight isomers. This indicates that (1S,3S,7R)-3-methyl-alpha-himachalene derived from L. longipalpis males may have a dual function in causing male aggregation as well as serving as a sex pheromone for females.     

Behavioural and electrophysiological responses of the phlebotomine sandfly Lutzomyia longipalpis (Diptera: Psychodidae) when exposed to canid host odour kairomones

Compounds from the odour-producing glands of the fox Vulpes vulpes were collected. This complex mixture of compounds was used to stimulate the ‘ascoid’ olfactory organs of female sandflies in single sensillum and gas chromatography-linked single sensillum recordings. Sixteen of these compounds were identified using gas chromatography-linked mass spectrometry and amounts present were determined. The compounds fell into four organic classes: ketones, carboxylic acids, alcohols and aldehydes. Specific neurones present in the ascoid sensillum that responded to each of these classes of compound were characterized. A bioassay chamber was developed that gave female sandflies the choice of two odour sources. Female sandflies were attracted upwind by fox odour and were trapped in closer proximity to the fox odour port than the control port. Synthetic compounds were recombined in appropriate quantities to mimic the fox odour. In this bioassay, the synthetic blend attracted sandflies upwind, and again they were caught closer to the test port than the control port. Furthermore, the synthetic fox odour induced an electrophysiological response from neurones in the ascoid sensillum that was very similar to that induced by natural fox odour. No synthetic compound alone induced the same behavioural response from sandflies as did whole fox odour. However, benzaldehyde, 4-hydroxy-4-methyl-2-pentanone and 4-methyl-2-pentanone alone did cause sandflies to fly upwind and to be caught closer to the test port than the control.     

Identification of Lutzomyia spp. (Diptera: Psychodidae) verrucarum group through electron microscopy of its eggs

The value of Colombian phlebotomine eggs for species determination was studied with a scanning electron microscope. The species diversity and medical importance of the verrucarum group were the bases to select Lutzomyia youngi, Lutzomyia evansi, Lutzomyia columbiana and Lutzomyia longiflocosa. The egg surface was poligonal. Lutzomyia youngi, and Lutzomyia columbiana had pentagonal or hexagonal patterns; Lutzomyia evansi elongated polygons and Lutzomyia longiflocosa irregular polygonal sculpturing, frequently rectangular. Egg scanning electron microscopy is reliable to identify species of the verrucarum group.     

Syntomoza matagalpensis,a New Moth-Fly from Nicaragua (Diptera,Psychodidae)

A new species of Psychodidae, Syntomoza matagalpensis from Nicaragua is described. Morphology of both male and female and notes on their biology are offered.