Microanatomical and secretory characterization of the salivary gland of the Rhodnius prolixus (Hemiptera, Reduviidae, Triatominae), a main vector of Chagas disease.

Rhodnius prolixus is the principal vector of Trypanosoma cruzi, the aetiological agent of Chagas disease in American countries. This insect is haematophagous during all life cycles and, to antagonize its haemostatic, inflammatory and immune systems, it secretes saliva while feeding on the vertebrate host's blood. Here, we investigated characteristic changes of the salivary glands (SG) that occur during insect development. Two pairs of lobules and ducts comprise the SG of R. prolixus. The organ's size increases over time, but the microanatomical structures are preserved during insect development. Both lobules have a single layer epithelium formed by binucleated cells, which surrounds the saliva reservoir. The principal lobule presents higher polysaccharide and total protein contents than the accessory lobe. A network of external muscle layers is responsible for organ contraction and saliva release. Apocrine, merocrine and holocrine secretion types occur in the secretory epithelium. Dopamine, serotonin and tyrosine-hydroxylase are neural-related molecules that regulate SG function both during and after feeding.

Morphological and ultrastructural analysis of an important place of sexual communication of Rhodnius prolixus (Heteroptera: Reduviidae): the Metasternal Glands.

Rhodnius prolixus is an important vector of Trypanosoma cruzi, the etiological agent of Chagas disease. Insect adults have a pair of Metasternal Glands (MGs) and the secretion emitted by these glands acts as sex pheromone. Recent studies have focused on the chemical composition of this pheromone, electrophysiological responses to MGs compounds and mating behavior assays. Morphological studies of these glands are still scarce. Thus, considering the relevance of MGs in the sex pheromone biosynthesis, we investigated the morphology and ultrastructure of R. prolixus MGs. The glandular apparatus presents a tubular structure containing secretory cells with canalicules that fuse with the central duct which conducts the secretion to a pear-shaped reservoir connected to the exterior by a droplet-shape orifice. The secretory cells are classified as class III, they present a well-developed rough and smooth endoplasmic reticulum. Smooth endoplasmic reticulum is a site of lipid biosynthesis that may be involved in the mevalonate pathway, a probable route of the sex pheromone biosynthesis in this insect. The presence of rough endoplasmic reticulum indicates a possible peptides/proteins secretions site which were still not characterized in MGs. Several mitochondria are scattered in the cytoplasm that may suggest a high metabolic activity. Further studies should be carried out to correlate these data with the sex pheromone biosynthesis in this vector.

Structure and function of the stylets of hematophagous Triatominae (Hemiptera: Reduviidae), with special reference to Dipetalogaster maxima.

Kissing bugs (Hemiptera: Reduviidae: Triatominae) are able to bend their rod-like maxillae while searching for blood vessels in the tissue of their vertebrate hosts. Little is known about the working mechanisms of these bending movements and the distal opening of the food channel. We compared the morphological structure of the stylets (mandibles and maxillae) of four triatomine species and analyzed the feeding process of Dipetalogaster maxima (Uhler, 1894). The maxillae of triatomine bugs are interlocked by a tongue-and-groove system, allowing longitudinal sliding. While penetrating the host tissue, the animals perform rapid alternate back and forth movements of the maxillae. The resistance of the surrounding tissue pushes the asymmetric apex of the maxillae away from its straight path, i.e., if one individual maxilla is protracted alone, its tip curves inwards, and the other maxilla follows. Once a blood vessel is tapped, the spine-like tip of the left maxilla splays outwards. Apically, each of the maxillae features an abutment, the left one exhibiting a notch that presumably facilitates splaying. The mechanical interaction of the two maxillary abutments enables the distal opening of the food channel but might also support the movements of the maxillary bundle attributable to different bending moment distributions.

[Blood-feeding of Rhodnius prolixus].

Triatomines (Hemiptera: Reduviidae) are blood-sucking insect vectors of the protozoan Trypanosoma cruzi which is the causative agent of Chagas' disease. Rhodnius prolixus is the most epidemiologically important vector of T. cruzi in Colombia. Triatomines are regarded to be vessel-feeders as they obtain their blood meals from vertebrate hosts by directly inserting their mouthparts into vessels. Microscopic techniques are useful for visualizing and describing the morphology of biological structures. Here, we show images of the blood-feeding of R. prolixus, including some histological features by light microscopy and scanning electron microscopy of the mouthparts of R. prolixus when feeding on a laboratory mouse.

Third record of Rhodnius amazonicus and comparative study with R. pictipes (Hemiptera, Reduviidae, Triatominae).

Rhodnius amazonicus Almeida et al. (1973) is a triatomine of rare occurrence. This species was found for the third time in Breves city, Pará state, Brazil. Morphometric and morphological studies were carried out on one male and one female. Lent and Wygodzinsky (1979) considered this species as a synonym of R. pictipes, until its revalidation after 23 years by Bérenger and Pluot-Sigwalt (2002). Considering the synonym mentioned above, a comparative study between these two species was performed in order to report the third encounter of this species, and increase the number of morphological characters that distinguish R. amazonicus from R. pictipes. The dorsal side of the head, the dorsal and ventral portions of the thorax, the dorsal, ventral and posterior sides of the female genitalia, eggs and the median process of the pygophore were examined by scanning electronic microscopy (SEM). The head, thorax, abdomen and egg parameters of these two species were also measured. The identification of characters on the head, stridulatory sulcus, mesosternum, metasternum, scutellum, process of urotergite I, external female genitalia, pygophore and eggs made the distinction between R. amazonicus and R. pictipes possible. This study has shown the new characters recorded and described for the first time for R. amazonicus and R. pictipes by SEM here made it possible to confirm the morphological separation between these two species. The morphometric analysis also confirmed that the above two taxa are different at specific level.

Aggregated oviposition in Rhodnius prolixus, sensory cues and physiological consequences.

Females of the haematophagous bug Rhodnius prolixus attach their eggs in clusters on substrates related to their hosts, such as nests or avian feathers. Because the hosts are an enormous food resource as well as potential predators, the choice of the site and pattern of oviposition could have an important adaptive value. Here we investigated proximate and a potential ultimate cause of this aggregated pattern of laid eggs. First, we studied proximal causes by analyzing the use of chemical or physical cues associated with aggregated oviposition in R. prolixus. For all terrestrial organisms there is a trade-off between exchange of respiratory gases and water loss. Particularly, insect eggs are highly susceptible to this trade-off because they do not obtain water from the environment, hence our second objective is to study the possible mechanisms involved in dehydration resistance in this species. Therefore we examined the dynamics of change in CO2 release rate (MCO2), and water loss rate (MH2O) in relation to embryo development as energetic demands increase, and tested the energetic or hygric efficiency hypothesis as a potential ultimate cause of aggregated oviposition. This hypothesis states that grouped eggs consume less energy or lose less water than equal numbers of isolated eggs, the latter being more susceptible to dehydration. Results indicated the use of physical external cues such as dummy eggs or edges of the oviposition substrates, but we did not find any chemical cues associated with the aggregated pattern of oviposition. There are no energetic or hygric benefits associated with egg's aggregated pattern. However, when we analyzed the MCO2 and MH2O change in relation to embryo development, we found a fairly constant and low MH2O albeit a clear increase in MCO2, suggesting a tightly control of egg's desiccation tolerance. This high resistance to desiccation coupled with a temporal strategy of hatching allows R. prolixus embryos to successfully develop and hatch under harsh environmental conditions.

Structure and postembryonic development of the intersegmental nodules in the non-muscular joints of the antennae in Rhodnius prolixus.

The antennae of Insecta consist of two basal segments and the distal annulated flagellum lacking intrinsic muscles. Non-muscular joints are important to preserve the flexibility and structure of the long heteropteran antennae which bear an intersegmental nodule on each non-muscular joint. Little is known about their properties or function. Here we characterize the structure and postembryonic development of the non-muscular joints of Rhodnius prolixus antennae. Using Scanning Electron Microscopy, we tracked the changes in shape and size of both intersegmental nodules during the course of the hemimetabolous insect life cycle. Using Atomic Force Microscopy, we established a qualitative correlation between the topography of the surface and the rigidity of the joint between pedicel and flagellum. Our results confirmed the presence of two sub-articulations on each non-muscular joint. Also, the two intersegmental nodules have different origins: the one between the two flagellar segments (intraflagelloid) is a sclerite already present from the early nymph, while the nodule between pedicel and flagellum (prebasiflagellite) originates by gradual separation of the proximal end of the basiflagellum during postembryonic development. Various changes occur in the non-muscular joints and segments of the antenna during the life cycle of R. prolixus.

Bitter stimuli modulate the feeding decision of a blood-sucking insect via two sensory inputs.

The gustatory system of animals is involved in food quality assessment and controls the feeding decision of an individual confronted with a potential alimentary source. Triatomines are haematophagous insects that feed on vertebrate blood. Once they reach a potential host, they walk over the host skin searching for an adequate site to pierce. Then, they insert their stylets and take a first sampling gorge to decide whether food is acceptable. Our work reveals that the presence of bitter compounds inhibits the feeding behavior of these bugs. Firstly, triatomines decreased their feeding behavior if substrates spread with quinine or caffeine were detected by external receptors localized exclusively in the antennae. Morphological inspections along with electrophysiological recordings revealed the existence of four gustatory sensilla located in the tip of the antenna that respond to both bitter tastants. The absence of these bitter detectors by antennal ablation reversed the observed feeding inhibition evoked by bitter compounds. Secondly, once triatomines pumped the first volume of food with bitter compounds (quinine, caffeine, berberine, salicin), a decrease in their feeding behavior was observed. Morphological inspections revealed the existence of eight gustatory sensilla located in the pharynx that might be responsible for the internal bitter detection. Finally, we found that a brief pre-exposure to bitter compounds negatively modulates the motivation of bugs to feed on an appetitive solution. Results presented here highlight the relevance of bitter taste perception in the modulation of the feeding behavior of a blood-sucking insect.

Study of the external female genitalia of 14 Rhodnius species (Hemiptera, Reduviidae, Triatominae) using scanning electron microscopy.

BACKGROUND: Among the vectors of Chagas disease (Hemiptera: Reduviidae:Triatominae), there are eighteen Rhodnius species described and some are difficult to identify. The aim of this article is to contribute to the specific identification of fourteen Rhodnius spp. through morphological characters of the external female genitalia. METHODS: Female abdomens were cut transversely. The specimens were then prepared for examination by using scanning electron microscopy. RESULTS: The careful examination of the dorsal, posterior and ventral sides revealed characteristics that allowed the identification of each of the fourteen species. CONCLUSION: The use of external female genitalia as characteristics are proposed as a tool for specifically identifying Rhodnius species, and an identification key for these species is presented.

Calreticulin expression levels and endoplasmic reticulum during late oogenesis and early embryogenesis of Rhodnius prolixus Stahl.

This study reports the cloning, expression analysis and localization of calreticulin (CRT) in the endoplasmic reticulum (ER) during late oogenesis and early embryogenesis of the insect Rhodnius prolixus. CRT was cloned and sequenced from cDNA extracted from unfertilized eggs. Real-time PCR showed that CRT expression remains at lower levels during late oogenesis when compared to vitellogenic oocytes or day 0 laid fertilized eggs. Immunofluorescence microscopy showed that this protein is located in the periphery of the egg, in a differential peripheral ooplasm surrounding the yolk-rich internal ooplasm, only identified by transmission electron microscopy (TEM) of thin sections. Using immunogold electron microscopy, the ER ultrastructure (CRT labeled) was identified in the peripheral ooplasm as dispersed lamellae, randomly distributed in the peripheral ooplasm. No massive alterations of ER ultrastructure were found before or right after (30 min) fertilization, but an increase in CRT expression levels and assembly of typical rough ER (parallel cisternae with associated ribosomes) were observed 18-24 h after oviposition. The lack of ER assembly at fertilization and the later formation of rough ER together with the increase in CRT expression levels, suggest that the major functions of ER might be of great importance during the early events of development. The possible involvement of ER in the early steps of embryogenesis will be discussed.

Characterization of the external female genitalia of six species of Triatominae (Hemiptera: Reduviidade) by scanning electron microscopy.

By macroscopic and microscopic dorsal side observation, it was noted that the IX and X segments of two species each of Panstrongylus and Triatoma terminate in an elongated way, whereas they terminate abruptly in the two species of Rhodnius. Scanning observation of the dorsal, ventral, lateral and posterior sides of the female genitalia of Panstrongylus herreri, Panstrongylus megistus, Rhodnius colombiensis, Rhodnius prolixus, Triatoma infestans and Triatoma vitticeps revealed that these six species are generally and specifically distinguished based on these elements. We describe several components that distinguish P. herreri from P. megistus: four on the dorsal side: the VII, VIII, IX and X segments, on the ventral view, three: VII sternite, VIII gonocoxite and VIII gonapophyse, on the lateral view one character, VIII gonocoxite and on the posterior view three characters: VIII and IX gonocoxite and XI gonopophyse. Comparing R. colombiensis and R. prolixus, there were three distinct characters on the dorsal side: the VII, VIII and X segments, on the ventral view three characters: the IX and X segments and VIII gonocoxite and on the posterior view four characters: the VIII, IX, X segments and VIII gonapophyse that distinguish the two species. T. infestans and T. vitticeps have four different characters on the dorsal side: the VII, VIII, IX and X segments, on the ventral view four characters: the VII and X segments, VIII gonocoxite and VIII gonapophyse, on the lateral view two characters, IX and X segments and on the posterior view four characters: the IX and X segments, VIII gonocoxite and VIII gonapophyse that distinguish the two species. Examination of the external female genitalia of six triatomine species by scanning suggests that these components are useful for taxonomical studies.

Characterization of the external female genitalia of six species of Triatominae (Hemiptera: Reduviidade) by scanning electron microscopy

By macroscopic and microscopic dorsal side observation, it was noted that the IX and X segments of two species each of Panstrongylus and Triatoma terminate in an elongated way, whereas they terminate abruptly in the two species of Rhodnius. Scanning observation of the dorsal, ventral, lateral and posterior sides of the female genitalia of Panstrongylus herreri, Panstrongylus megistus, Rhodnius colombiensis, Rhodnius prolixus, Triatoma infestans and Triatoma vitticeps revealed that these six species are generally and specifically distinguished based on these elements. We describe several components that distinguish P. herreri from P. megistus: four on the dorsal side: the VII, VIII, IX and X segments, on the ventral view, three: VII sternite, VIII gonocoxite and VIII gonapophyse, on the lateral view one character, VIII gonocoxite and on the posterior view three characters: VIII and IX gonocoxite and XI gonopophyse. Comparing R. colombiensis and R. prolixus, there were three distinct characters on the dorsal side: the VII, VIII and X segments, on the ventral view three characters: the IX and X segments and VIII gonocoxite and on the posterior view four characters: the VIII, IX, X segments and VIII gonapophyse that distinguish the two species. T. infestans and T. vitticeps have four different characters on the dorsal side: the VII, VIII, IX and X segments, on the ventral view four characters: the VII and X segments, VIII gonocoxite and VIII gonapophyse, on the lateral view two characters, IX and X segments and on the posterior view four characters: the IX and X segments, VIII gonocoxite and VIII gonapophyse that distinguish the two species. Examination of the external female genitalia of six triatomine species by scanning suggests that these components are useful for taxonomical studies.

Ecdysteroid receptor (EcR) is associated with microtubules and with mitochondria in the cytoplasm of prothoracic gland cells of Rhodnius prolixus (Hemiptera).

We have shown previously that EcR in larval Rhodnius is present in the cytoplasm of various cell types and undergoes daily cycling in abundance in the cytoplasm (Vafopoulou and Steel, 2006. Cell Tissue Res 323:443-455). It is unknown which organelles are associated with EcR. Here, we report that cytoplasmic EcR in prothoracic gland cells is associated with both microtubules and mitochondria, and discuss the implications for both nuclear and non-genomic actions of EcR. EcR was localized immunohistochemically using several antibodies to EcR of Manduca and Drosophila and a confocal laser scanning microscope. Double labels were made to visualize EcR and (1) microtubules (using an antibody to tyrosylated alpha-tubulin) and (2) mitochondria (using a fluorescent MitoTracker probe), both after stabilization of microtubules with taxol. EcR co-localized with both tubulin and mitochondria. All the different EcR antibodies produced similar co-localization patterns. EcR was seen in the perinuclear aggregation of mitochondria, indicating that mitochondria are targets of ecdysone, which could influence mitochondrial gene transcription. EcR was also distributed throughout the microtubule network. Co-localization of EcR with tubulin or mitochondria was maintained after depolymerization of microtubules with colchicine. Treatment with taxol resulted in accumulation of EcR in the cytoplasm and simultaneous depletion of EcR from the nucleus, suggesting that microtubules may be involved in targeted intracellular transport of EcR to the nucleus (genomic action) or may play a role in rapid ecdysone signal transduction in the extranuclear compartment, i.e., in non-genomic actions of ecdysone. These findings align EcR more closely with steroid hormone receptors in vertebrates.

Antiserum against perimicrovillar membranes and midgut tissue reduces the development of Trypanosoma cruzi in the insect vector, Rhodnius prolixus.

Antiserum raised against Rhodnius prolixus perimicrovillar membranes (PMM) and midgut tissue interfered with the midgut structural organization and reduced the development of Trypanosoma cruzi in the R. prolixus insect vector. SDS-PAGE and Western blot analyses confirmed the specific recognition of midgut proteins by the antibody. Feeding, mortality, molt, and oviposition of the insects were unaffected by feeding with the antiserum. However, the eclosion of the eggs were reduced from R. prolixus females treated with antiserum. Additionally, in vivo evaluation showed that after oral treatment with the antiserum, the intensity of infection with the Dm-28c clone of T. cruzi decreased in the digestive tract of fifth-instar nymphs and in the excretions of R. prolixus adults. These results suggest that the changes observed in the PMM organization in the posterior midgut of R. prolixus may not be important for triatomine survival but the antiserum acts as a transmission-reduction vaccine able to induce significant decreases in T. cruzi infection in the vector.

The haemoxisome: a haem-iron containing structure in the Rhodnius prolixus midgut cells.

Rhodnius prolixus midgut was analysed using transmission electron microscopy and electron spectroscopic imaging in order to localize the cellular structures involved in haem metabolism. In the posterior midgut, special cellular electron-dense structures were observed. These structures are here designated haemoxisomes. Haemoxisomes are present in the epithelial cells at various time points after a blood meal. Several days after the blood meal, some of them become less electron-dense. By electron spectroscopic imaging, large amounts of iron and oxygen were detected in these cellular structures. The iron is probably bound to the porphyrin ring as an iron-protoporphyrin IX complex, as detected using the diaminobenzidine technique. An interesting observation was the presence of endoplasmic reticulum surrounding the haemoxisomes during some special periods. Iron content was monitored in the posterior midgut epithelium and was found to be constant at the initial days after a blood meal, but slightly higher at the end of the digestive process (from 13th up to 20th day). These results are in agreement with the observation that the appearance of the haemoxisomes changes at the end of the digestive process. The ability to degrade haem seems to depend on the presence of endoplasmic reticulum as observed using a haem degradation assay in the presence of an endoplasmic reticulum-enriched fraction. Taken together these results suggest that haemoxisomes may play a role in intracellular haem detoxification.

Morphobiological aspects of Rhodnius brethesi Matta, 1919 ( Hemiptera:Reduviidae ) from the Upper and Middle Negro River, Amazon region of Brazil: I - scanning electron microscopy

The occurrence of autochthonous cases of Chagas disease in the Amazon region of Brazil over recent decades has motivated an intensification of studies in this area. Different species of triatomines have been identified, and ten of these have be proven to be carriers of the parasite Trypanosoma cruzi or " cruzi-like " parasites. Studies conducted in the municipalities of Santa Isabel do Rio Negro and Barcelos, located on the Upper and Middle of the Negro River, microregion of Negro River, state of Amazonas have confirmed not only that Rhodnius brethesi is present in the palm tree Leopoldinia piassaba, but also that this insect was recognized by palm fiber collectors. A morphological study of eyes, inter-ocular and inter-ocellar regions, antennae, buccula, labrum, rostrum, stridulatory sulcus and feet, including the apex of the tibia, spongy fossette and ctenidium was conducted by scanning electron microscopy. The buccula and the stridulatory sulcus presented notable differences in specimens of different genera and also of different species. These data make it possible to suggest that the details presented in these structures can be included as diagnostic characteristics to be used in new dichotomous keys, thereby contributing towards studies of taxonomy and systematics and furnishing backing for comparative analysis of specimens collected from different localities.

Morphobiological aspects of Rhodnius brethesi Matta, 1919 (Hemiptera:Reduviidae) from the Upper and Middle Negro River, Amazon region of Brazil: I - scanning electron microscopy.

The occurrence of autochthonous cases of Chagas disease in the Amazon region of Brazil over recent decades has motivated an intensification of studies in this area. Different species of triatomines have been identified, and ten of these have be proven to be carriers of the parasite Trypanosoma cruzi or " cruzi-like " parasites. Studies conducted in the municipalities of Santa Isabel do Rio Negro and Barcelos, located on the Upper and Middle of the Negro River, microregion of Negro River, state of Amazonas have confirmed not only that Rhodnius brethesi is present in the palm tree Leopoldinia piassaba, but also that this insect was recognized by palm fiber collectors. A morphological study of eyes, inter-ocular and inter-ocellar regions, antennae, buccula, labrum, rostrum, stridulatory sulcus and feet, including the apex of the tibia, spongy fossette and ctenidium was conducted by scanning electron microscopy. The buccula and the stridulatory sulcus presented notable differences in specimens of different genera and also of different species. These data make it possible to suggest that the details presented in these structures can be included as diagnostic characteristics to be used in new dichotomous keys, thereby contributing towards studies of taxonomy and systematics and furnishing backing for comparative analysis of specimens collected from different localities.

Effect of blood components, abdominal distension, and ecdysone therapy on the ultrastructural organization of posterior midgut epithelial cells and perimicrovillar membranes in Rhodnius prolixus.

The effects of blood components, nerve-cord severance, and ecdysone therapy on the posterior midgut epithelial cells of 5th-instar Rhodnius prolixus nymphs 10 days after feeding were analyzed by transmission electron microscopy. Cutting the nerve-cord of the blood-fed insects partially reduced the development of microvilli and perimicrovillar membranes (PMM), and produced large vacuoles and small electrondense granules; insects fed on Ringer's saline diet exhibited well developed microvilli and low PMM production; swolled rough endoplasmatic reticulum and electrondense granules; Ringer's saline meal with ecdysone led to PMM development, glycogen particles, and several mitochondria in the cytoplasm; epithelial cells of the insects fed on Ringer's saline meal whose nerve-cord was severed showed heterogeneously distributed microvilli with reduced PMM production and a great quantity of mitochondria and glycogen in the cytoplasm; well developed microvilli and PMM were observed in nerve-cord severed insects fed on Ringer's saline meal with ecdysone; Ringer's saline diet containing hemoglobin recovered the release of PMM; and insects fed on human plasma showed slightly reduced PMM production, although the addition of ecdysone in the plasma led to a normal midgut ultrastructural organization. We suggest that the full development of microvilli and PMM in the epithelial cells depends on the abdominal distension in addition to ingestion of hemoglobin, and the release of ecdysone.

Ultrastructure of the salivary glands of Rhodnius domesticus Neiva & Pinto, 1923 (Hemiptera: Reduviidae).

The fine structure of the salivary glands of the triatomine bug Rhodnius domesticus was investigated. Stereomicroscopy and scanning electron microscopy showed that each salivary gland pair contains two close and independent units: the larger is reddish and elongated (principal gland), while the smaller is round and translucent (accessory gland). The accessory gland opens at the base of the main excretion duct, which arises at the medial portion of the principal gland. An accessory duct emerges at the base of the main excretion duct, above the accessory gland opening, and runs towards the digestive tract. Transmission electron microscopy showed that both gland units are formed by a single layer of epithelial gland cells, surrounded by a thick basal lamina containing tracheolae and muscle cell fibers. Adjacent gland cells are interconnected by interdigitations of their lateral plasma membranes and by septate junctions. Microvilli are present at the apical domain of the gland cell plasma membrane, which allow faster diffusion of the saliva towards the gland lumen. Several mitochondria, abundant endoplasmic reticulum profiles and usually one elongated nucleus are observed in the gland cells. According to standard nomenclatures, the salivary gland cells can be classified as type I cells, secreting the saliva into a large gland lumen.

Inter-specific and developmental differences on the array of antennal chemoreceptors in four species of Triatominae (Hemiptera: reduviidae).

The aim of the work was to investigate the pattern of chemoreceptor sensilla in adults and fifth stage nymphs of Rhodnius prolixus, R. neglectus, Triatoma infestans and T. sordida in order to study differences and similarities between genera and species. Three types of sensilla were analyzed by light microscopy: thin-walled trichoidea, thick-walled trichoidea and basiconica. The number of sensilla of each three types were counted. The length of the antennal segments were also used as a variable for the analysis. The statistical analysis showed that the number of these antennal chemoreceptors had significant differences between species and between adults and nymphs of each species. Discriminant analysis separates incompletely the fifth stage nymphs of the four species and showed similarity between them. Discriminant analysis performed with 12 variables of the antennae, allowed a complete separation of the adults of the four species.