Preparation of sand fly (Diptera: Psychodidae: Phlebotominae) specimens for histological studies

Phlebotominae sand fly specimens were prepared for histological and physiological studies. Different fixatives were tested on sectioned and whole bodied adult females in order to obtain good fixation and provide satisfactory penetration of the embedding media. All fixed specimens were infiltrated (up to seven days under 5ºC) and embedded in hydroxyethyl metacrylate. Two-three µm sections were stained, mounted in Canada balsam and observed by light microscopy. Best results were achieved when whole bodied insects were double fixed in Bouin's and Carnoy's fluids (4 h/2 h) and stained in Hematoxilin/Eosin or fixed in calcium formaldehyde and stained in mercury bromophenol blue.

Description of Simulium damascenoi (Diptera: Simuliidae) male and the black-fly species from the State of Amapá, Brazil

Five species are included in the Simulium siolii group, which is placed in the subgenus Psaroniocompsa (Diptera: Simuliidae). Of these five species, only two (Simulium siolii Py-Daniel and Simulium tergospinosum Hamada) have been described in all their life stages, except eggs. Knowledge of the taxonomic characters of all life stages of a species is important in order to clarify interspecific and higher-level taxonomic relationships. The objectives of the present study are to describe the male of Simulium damascenoi Py-Daniel, to provide a list of black-fly species their bionomics and distributions in the state of Amapá, Brazil, and to provide an identification key for larvae and pupae for these species.

Triatoma rubrovaria (Blanchard, 1843) (Hemiptera-Reduviidae-Triatominae) III: patterns of feeding, defecation and resistance to starvation

Data from the Chagas Disease Control Program indicate a growing domiciliary and peridomiciliary invasion of Triatoma rubrovaria in the State of Rio Grande do Sul, where it has become the most frequent triatomine species captured there since the control of T. infestans. Bionomic characteristics that could influence the vectorial capacity of T. rubrovaria as vector of Trypanosoma cruzi were evaluated: patterns of (i) feeding, (ii) defecation, and (iii) resistance to starvation, using insects fed on mice. Fifty three percent of the females showed a defecation pattern conducive to chagasic transmission, defecating either on or near the bite site. The averages of the resistance to starvation varied from 48.1 to 179 days, for the first and fifth nymphal stages, respectively. Our study shows that with respect to the patterns of feeding, defecation and resistance to fasting, T. rubrovaria presented similar rates to the ones observed for other effective vector species, such as T. infestans. Thus, based on our studies we conclude that T. rubrovaria has biological characteristics that can positively influence its capacity to become infected and transmit T. cruzi, and also to keep residual populations after chemical control interventions.

Larval biometry of Simulium rubrithorax (Diptera: Simuliidae) and size comparison between populations in the states of Minas Gerais and Roraima, Brazil

The number of larval instars of Simulium (Hemicnetha) rubrithorax Lutz (Diptera: Nematocera) was determined using the lateral length of the head capsule. In this study 1,035 larvae, of different sizes, were measured (639 from the state of Roraima and 396 from the state of Minas Gerais). A frequency distribution analysis was carried out on the measurements of the lateral length of the head capsule to determine the number of larval instars. The limits of each instar were defined by the lower frequency of the measurements falling in a range of values, by the presence of the "egg burster" that characterizes the first larval instar, and by the developmental stage of the gill histoblast. The determination of the instar number was tested using a Student's t-test (p < 0.05), the Dyar rule and the Crosby growth rule. The results indicate the existence of 7 larval instars for this species, although this result was not in accordance to the Crosby rule. Last-instar larvae from two widely separated geographical populations (Roraima and Minas Gerais), collected in habitats with different water temperature were compared and no differences (p > 0.05) were observed between them.

Development of genital plates in nymphs of Triatoma pallidipennis, Stål 1872, (Hemiptera: Reduviidae) and its application as sexing method

Searching for morphometric differences between sexes in immature forms, the development of genital plates in the exuviae of Triatoma pallidipennis Stål 1872nymphs was studied. Differences were found in the form and size of the 9th genital urosternite, it is larger and wider in males as compared to females. This difference is reported in several South American Triatoma species. From our results it is possible to sex early stages from microscopic observation of genital plates in whole insects.

Foco de doença de Chagas em Arcádia, estado do Rio de Janeiro, Brasil

Following the report of triatomine nymphs in a house in Arcadia, Miguel Pereira, state of Rio de Janeiro, Brazil, the infested dwelling was checked. Several eggs and 46 specimens of Triatoma vitticeps (Stal, 1859) were collected. Among them, adults and nymphal instars accounted for 43.5% and 56.5%, respectively. Analysis of blood meals showed the ecletism of this species; 24 (52.2%) were single feeds, 18 insects (39.1%) fed on two hosts and 4 (8.7%) on three hosts. Trypanosoma cruzi infection rate of examined specimens was 13%. Finally one of the residents of the house was positive for anti-T. cruzi antibodies using indirect immunofluorescence.

Flight metabolism in Panstrongylus megistus (Hemiptera: Reduviidae): the role of carbohydrates and lipids

The metabolism of lipids and carbohydrates related to flight activity in Panstrongylus megistus was investigated. Insects were subjected to different times of flight under laboratory conditions and changes in total lipids, lipophorin density and carbohydrates were followed in the hemolymph. Lipids and glycogen were also assayed in fat body and flight muscle. In resting insects, hemolymph lipids averaged 3.4 mg/ml and significantly increased after 45 min of flight (8.8 mg/ml, P < 0.001). High-density lipophorin was the sole lipoprotein observed in resting animals. A second fraction with lower density corresponding to low-density lipophorin appeared in insects subjected to flight. Particles from both fractions showed significant differences in diacylglycerol content and size. In resting insects, carbohydrate levels averaged 0.52 mg/ml. They sharply declined more than twofold after 15 min of flight, being undetectable in hemolymph of insects flown for 45 min. Lipid and glycogen from fat body and flight muscle decreased significantly after 45 min of flight. Taken together, the results indicate that P. megistus uses carbohydrates during the initiation of the flight after which, switching fuel for flight from carbohydrates to lipids.

Hemilucilia segmentaria (Fabricius, 1805) (Diptera: Calliphoridae) as new biological vector of eggs of Dermatobia hominis (Linnaeus Jr., 1781) (Diptera: Oestridae) in Reserva Biológica do Tinguá, Rio de Janeiro, Brazil

The aim of this note was to record for the first time the finding of Hemilucilia segmentaria acting as biological vector of Dermatobia hominis, during a study of the diversity of Calliphoridae at Reserva Biológica do Tinguá, Rio de Janeiro, Brazil. The insects were captured using traps baited with chicken vicera, for a period of 28-30 h twice per month. In the period of one year, 1987 insects were collected, 7.5% of which belonged to the H. segmentaria; of these a female was captured in May 2001, carrying a mass of 20 eggs on the left side of its abdomen.

Task-dependent influence of genetic architecture and mating frequency on division of labour in social insect societies.

Division of labour is one of the most prominent features of social insects. The efficient allocation of individuals to different tasks requires dynamic adjustment in response to environmental perturbations. Theoretical models suggest that the colony-level flexibility in responding to external changes and internal perturbation may depend on the within-colony genetic diversity, which is affected by the number of breeding individuals. However, these models have not considered the genetic architecture underlying the propensity of workers to perform the various tasks. Here, we investigated how both within-colony genetic variability (stemming from variation in the number of matings by queens) and the number of genes influencing the stimulus (threshold) for a given task at which workers begin to perform that task jointly influence task allocation efficiency. We used a numerical agent-based model to investigate the situation where workers had to perform either a regulatory task or a foraging task. One hundred generations of artificial selection in populations consisting of 500 colonies revealed that an increased number of matings always improved colony performance, whatever the number of loci encoding the thresholds of the regulatory and foraging tasks. However, the beneficial effect of additional matings was particularly important when the genetic architecture of queens comprised one or a few genes for the foraging task's threshold. By contrast, a higher number of genes encoding the foraging task reduced colony performance with the detrimental effect being stronger when queens had mated with several males. Finally, the number of genes encoding the threshold for the regulatory task only had a minor effect on colony performance. Overall, our numerical experiments support the importance of mating frequency on efficiency of division of labour and also reveal complex interactions between the number of matings and genetic architecture.

Simulium (Chirostilbia) bifenestratum (Diptera, Simuliidae), a new black-fly species from the Atlantic forest, State of São Paulo, Brazil

The larva, pupa, male, and female of Simulium bifenestratum n. sp. are described and illustrated. The pupae of the new species have 10 gill filaments, thick at their base and arranged in a three-dimensional way, surrounding the head and thorax. Its pupal cocoon is peculiar, not found in any of the known Brazilian black-fly species; it is very thick and hard with two openings in the anterior region. S. bifenestratum n. sp. was collected in one stream in the Bocaina mountain chain, Atlantic forest, in São José do Barreiro county, state of São Paulo, in a high (1500 m) natural grassland. Larvae and pupae were collected on the edges of small waterfalls and in places with-high speed laminar water flow, attached to the bedrock.

Differential gene expression in response to mechanical wounding and insect feeding in Arabidopsis.

Wounding in multicellular eukaryotes results in marked changes in gene expression that contribute to tissue defense and repair. Using a cDNA microarray technique, we analyzed the timing, dynamics, and regulation of the expression of 150 genes in mechanically wounded leaves of Arabidopsis. Temporal accumulation of a group of transcripts was correlated with the appearance of oxylipin signals of the jasmonate family. Analysis of the coronatine-insensitive coi1-1 Arabidopsis mutant that is also insensitive to jasmonate allowed us to identify a large number of COI1-dependent and COI1-independent wound-inducible genes. Water stress was found to contribute to the regulation of an unexpectedly large fraction of these genes. Comparing the results of mechanical wounding with damage by feeding larvae of the cabbage butterfly (Pieris rapae) resulted in very different transcript profiles. One gene was specifically induced by insect feeding but not by wounding; moreover, there was a relative lack of water stress-induced gene expression during insect feeding. These results help reveal a feeding strategy of P. rapae that may minimize the activation of a subset of water stress-inducible, defense-related genes.

Feeding sources and natural infection of Belminus herreri (Hemiptera, Reduviidae, Triatominae) from dwellings in Cesar, Colombia

Belminus herreri, originally described from specimens collected in Panama, was considered entirely silvatic until to 2000 when it was found for the first time in a domestic habitat in Colombia. In 2001, during a new search of houses in the Department of Cesar, Colombia, 121 specimens were collected. Study of their feeding sources using an ELISA test revealed that 96% of these specimens had fed on cockroaches (Blattidae). However, a small proportion of these B. herreri specimens also showed the presence of Trypanosoma cruzi in their gut contents, suggesting a possible role for these insects in the epidemiology of Chagas disease.

Activity of oil-formulated Beauveria bassiana against Triatoma sordida in peridomestic areas in central Brazil

Field tests were carried out during the rainy season of 2001/2002 in São Luís de Montes Belos, Goiás, Brazil, to evaluate the potential of the entomopathogenic fungus, Beauveria bassiana, against peridomestic Triatoma sordida. An oil-water formulation of the isolate CG 14 (Embrapa) was applied in triatomine infested hen houses of four farms at a final concentration of 10(6) conidia/cm². Numbers of T. sordida decreased over the next 25 days, after application of the fungus, and B. bassiana developed on dead insects in one hen house. A high number of B. bassiana colonies was detected in substrates collected in treated hen houses 24 h after application of CG 14. In the following three months the presenceof B. bassiana declined to values found before treatment.

Role of enhanced detoxication in a deltamethrin-resistant population of Triatoma infestans (Hemiptera, Reduviidae) from Argentina

Deltamethrin and other pyrethroids have been extensively used in Argentina since 1980, for the chemical control of Triatoma infestans Klug (Hemiptera: Reduviidae). Recently, resistance to deltamethrin was detected in field populations by the survival of bugs exposed by topical application to the diagnostic dose estimated on the CIPEIN susceptible strain. Results of the current study showed low resistant ratios (RRs) to deltamethrin for the resistant populations (RR ranged from 2.0 for San Luis colony to 7.9 for Salta colony). Biochemical studies were made on the most resistant colony (Salta) and the susceptible strain (CIPEIN), in order to establish the importance of degradative mechanisms as a cause of the detected resistance. Esterase activity was measured on 3 days old first instars through phenylthioacetate and a-naphtyl acetate activities. The results showed a significant difference in no cholinesterase esterase activity from susceptible (7.6 ± 0,7 µM S./i.min.) and Salta resistant colony (9.5 ± 0.8 µM S./i.min.). Cytochrome P450 mono-oxygenase (P450) activity was measured on individual insects through ethoxycoumarine deethylase (ECOD) activity using a fluorescence micro plate reader. The dependence of ECOD activity on age and body region of the nymphs, and pH and time of incubation were studied in order to optimize the measurement. As a result, comparative studies were performed on abdomens of 2 days old first instars at pH 7.2 and 4 h incubation time. ECOD activity of first nymphs was significantly lower in the susceptible colony (61.3 ± 9.08 pg ECOD/ insect) than in the resistant one (108.1± 5.7 pg ECOD/ insect). These results suggest that degradative esterases (no-cholinesterase) and mono-oxygenases cytochrome P450, play an important role in the resistance to deltamethrin in Salta colony from Argentina.

Insecticide and community interventions to control Triatoma dimidiata in localities of the State of Veracruz, Mexico

Three different interventions to control Triatoma dimidiata in the State of Veracruz were implemented: X-1 = whole dwelling spraying, X-2 = middle wall spraying, X-3 = household cleaning. Cyfluthrin was sprayed 3 times with 8 month intervals. After each spraying, insects were collected and sent to the laboratory to be recorded and to determine genus and species of the adult triatomine bugs, and nymphs were counted. Trypanosoma cruzi presence was determined. With X-1, the infestation, colonization, and natural infection indexes were reduced to 0% in the 3 localities, with respect to t0. With X-2, the infestation index was reduced to 10% at t3 in 3 localities; the colonization index was reduced to 0% in only 1 locality at t3, and the natural infection index was reduced to 0% at t3. With X-3 the 3 indexes were not effectively reduced but they decreased with respect to the baseline study. Insecticide application to the whole dwelling is a more efficient intervention than its application to only the lower half of the walls and to the cleaning of houses.