Prevalence and intensity of infection, metacyclogenesis and nuclear phenotypes in Panstrongylus megistus (Burmeister, 1835) after ingestion of Trypanosoma cruzi (Chagas, 1909) II and subjection to heat shock.

This study aimed to contribute to our knowledge of the parasite-vector interaction associated with Trypanosoma cruzi (Chagas, 1909) infection in Panstrongylus megistus (Burmeister, 1835), an important vector of Chagas' disease in Brazil. The prevalence and intensity of T. cruzi infection, the incidence of metacyclogenesis and the frequency of nuclear phenotypes in Malpighian tubules were investigated in nymphs of P. megistus, reared at 28 °C and subjected to heat shock (40 °C, 1 hour) two days after infection with T. cruzi II (Y strain). Following the 45-day post-infection period, the frequency of epimastigotes was much higher than that of trypomastigotes in both heat-shocked and non-shocked insects, and the prevalence of infection was not altered by heat shock. Fewer epimastigotes and trypomastigotes were found in the infected insects subjected to the heat shock, indicating that the multiplication and metacyclogenesis of the parasites were affected by the stress. In infected specimens heat shock promoted an increased frequency of cell nuclei with heterochromatin decondensation, a cell survival response to stress, and did not affect insect survival. The effects of infection and heat shock, especially on the multiplication and metacyclogenesis of T. cruzi, and the observed resistance to heat shock developed by P. megistus nymphs are suggestive that they should be considered when adequate conditions for rearing these infected insects in the laboratory are pursued.

Eye pigments of the blood-sucking insect, Triatoma infestans Klug (Hemiptera, Reduviidae).

The pigmentation of black (wild) and red (mutant) eyes of Triatoma infestans was studied spectrophotometrically and compared with red-eyed (wild) and white-eyed (mutant) forms of Drosophila melanogaster. The spectral absorption profiles of the black and red eye pigments of T. infestans were similar to each other and to that of the wild-type eyes of D. melanogaster. The similarity to the wild form of D. melanogaster indicated that both eye forms of T. infestans contained ommochromes of the xanthommatin type, a finding confirmed by ascending paper chromatography. Pteridines, melanins, and ommins were not detected as eye pigments in T. infestans. The eye color difference in T. infestans was assumed to be a function of the xanthommatin concentration, with a smaller content of ommochrome in red eyes, although this probably did not affect the insect's visual acuity. These data support other findings regarding the similarities between black- and red-eyed specimens of T. infestans for other characteristics.

Eye pigments of the blood-sucking insect, Triatoma infestans klug (Hemiptera, Reduviidae)

A pigmentação de olhos pretos (forma selvagem) e vermelhos (forma mutante) de Triatoma infestans foi estudada por espectrofotometria e comparada à de olhos vermelhos (selvagem) e brancos (mutante) de Drosophila melanogaster. Os perfis do espectro de absorção dos pigmentos de olho preto e vermelho de T. infestans foram semelhantes entre si e ao dos olhos de tipo selvagem de D. melanogaster. A similaridade com a forma selvagem de D. melanogaster indicou que ambos os tipos de olho de T. infestans continham omocromos do tipo xantomatina, o que foi confirmado por cromatografia ascendente em papel. Não foram detectadas pteridinas, melaninas e ominas como pigmentos de olho em T. infestans. A diferença na cor de olho em T. infestans foi considerada uma função da concentração de xantomatina, sendo menor o conteúdo de omocromo nos olhos vermelhos, embora isso provavelmente não afete a acuidade visual do inseto. Esses resultados estão de acordo com dados de outros autores quanto a semelhanças envolvendo outras características entre espécimes de olho preto e vermelho de T. infestans.

Experimentally induced heat- and cold-shock tolerance in adult Panstrongylus megistus (Burmeister) (Hemiptera, Reduviidae)

The survival rate of domestic male and female adult Panstrongylus megistus was studied after sequential heat and cold shocks in order to investigate shock tolerance compared to that previously reported for nymphs. Sequential shocks were such that a milder shock (0ºC, 5ºC, 35ºC, or 40ºC for 1 h) preceded a severe one (0ºC or 40ºC for 12 h), separated by intervals of 8, 18, 24, and 72 h at 28ºC (control temperature). The preliminary thermal shock induced tolerance to the more severe one, although tolerance intensity depended on the initial shock temperature and the interval between treatments. Despite the observed tolerance, the survival rate for insects subjected to both shocks decreased when compared to that of individuals subjected to a single mild shock. When tolerance differed with sex, females showed greater values than males. In contrast to the response detected in nymphs, for which higher heat tolerance values were sustained for intervals of up to 24 h (preliminary shock, 35ºC) or even longer (preliminary shock, 40ºC) between sequential shocks, significant values were verified in adults only for shock intervals of up to 8 h (preliminary shock, 40ºC). While findings for nymphs exhibited considerable cold-shock tolerance under conditions in which preliminary shocks were given at 5ºC or 0ºC and the periods between shocks were up to 72 h long, the adults were shown to be capable of acquiring a substancial tolerance response to a more severe cold shock only when the preliminary shock was given at 0ºC and shock interval surpassed 18 h. It is assumed that the mechanisms involved in the cellular protection of P. megistus under sequential temperature shocks (heat shock protein action?) may loose effectiveness with insect development.

Experimentally induced heat- and cold-shock tolerance in adult Panstrongylus megistus (Burmeister) (Hemiptera, Reduviidae).

The survival rate of domestic male and female adult Panstrongylus megistus was studied after sequential heat and cold shocks in order to investigate shock tolerance compared to that previously reported for nymphs. Sequential shocks were such that a milder shock (0 degree C, 5 degrees C, 35 degrees C, or 40 degrees C for 1 h) preceded a severe one (0 degree C or 40 degrees C for 12 h), separated by intervals of 8, 18, 24, and 72 h at 28 degrees C (control temperature). The preliminary thermal shock induced tolerance to the more severe one, although tolerance intensity depended on the initial shock temperature and the interval between treatments. Despite the observed tolerance, the survival rate for insects subjected to both shocks decreased when compared to that of individuals subjected to a single mild shock. When tolerance differed with sex, females showed greater values than males. In contrast to the response detected in nymphs, for which higher heat tolerance values were sustained for intervals of up to 24 h (preliminary shock, 35 degrees C) or even longer (preliminary shock, 40 degrees C) between sequential shocks, significant values were verified in adults only for shock intervals of up to 8 h (preliminary shock, 40 degrees C). While findings for nymphs exhibited considerable cold-shock tolerance under conditions in which preliminary shocks were given at 5 degrees C or 0 degree C and the periods between shocks were up to 72 h long, the adults were shown to be capable of acquiring a substantial tolerance response to a more severe cold shock only when the preliminary shock was given at 0 degree C and shock interval surpassed 18 h. It is assumed that the mechanisms involved in the cellular protection of P. megistus under sequential temperature shocks (heat shock protein action?) may loose effectiveness with insect development.

[Triatoma infestans in area under entomological surveillance for Chagas' disease in São Paulo State, Brazil]. / Triatoma infestans em área sob vigilância entomológica para doença de Chagas, Estado de São Paulo, Brasil.

In response to notification of the capture of a winged triatomine bug by local inhabitants, an epidemiological investigation was investigated in the rural area of the Municipality of Paulínia, São Paulo State (Brazil). This led to the collection of 109 specimens of Triatoma infestans from peridomiciliary breeding sites. Local conditions favored colonization by triatomine bugs: many abandoned outbuildings were inhabited by pigeons and sparrows, which meant abundant feed for these bugs. Thus, surveillance is mandatory as long as conditions for triatomine breeding persist. Nevertheless, entomological and serological indicators ruled out immediate concern over vectorial transmission of Chagas' disease in this municipality. All domiciliary units of the region were sprayed with residual-effect insecticides and the triatomine breeding sites were destroyed by the use of mechanical control of bird nests. The importance of reporting the occurrence of triatomine bugs, particularly Triatoma infestans, in this region, is emphasized.

Experimentally induced heat-shock tolerance in Panstrongylus megistus (Hemiptera: Reduviidae).

The survival and molting incidence of fifth-instar nymphs of Panstrongylus megistus (Burmeister, 1835), a vector of Chagas' disease, were investigated following sequential heat shocks in which a mild shock (35 or 40 degrees C, 1 h) preceded a more drastic one (40 degrees C, 12 h). The shocks were separated by 8-, 18-, 24-, or 72-h periods at 28 degrees C. The heat-shock tolerance response was more effective when the first shock was given at 40 degrees C. When the period between shocks was 18 h, the tolerance to sequential shocks (in terms of specimen survival) weakened, which suggested a transient control of the process that enables the organism to circumvent the unfavorable effects of severe shock. In terms of molting incidence, the heat-shock tolerance was only demonstrated when the period between the first shock at 40 degrees C for 1 h and the second shock at 40 degrees C for 12 h was > or = 24 h. These results are the first to demonstrate the induction of heat-shock tolerance in a blood-sucking hemipteran.

Effect of sequential cold shocks on survival and molting incidence in Panstrongylus megistus (Burmeister) (Hemiptera, Reduviidae).

The survival and molting incidence were studied in the insect, Panstrongylus megistus, following sequential cold shocks in which a milder shock at 0 or 5 degrees C for 1 h preceded a more severe shock (0 degrees C, 12 h). The shocks were separated by intervals of 8, 18, 24, and 72 h at 28 degrees C. The survival rate after sequential shocks was identical to that of unshocked controls. Cold-shock tolerance differed from heat-shock tolerance since the latter varied with the time between shocks and was much more transient. Sequential cold shocks produced a higher molting incidence when the first shock was given at 0 compared to 5 degrees C. This response was more rapid than that to sequential heat shocks. Cold-shock tolerance in P. megistus may involve heat-shock proteins, although other protective mechanisms may also occur concurrently. This is the first report of cold-shock tolerance in a blood-sucking hemipteran.

Cell death and survival alterations in Malpighian tubules of Triatoma infestans following heat shock.

In this study, we examined cell survival and cell death in response to heat shock in an insect organ composed of highly polyploid cells no longer capable of cell division. For this, the frequency of nuclear phenotypes in Feulgen-stained Malpighian tubules of the blood-sucking insect, Triatoma infestans, was analyzed at various times after a short heat shock with or without subsequent moderate fasting. Cell death DNA fragmentation was studied immunocytochemically. Normal phenotypes and phenotypes indicative of cell survival (heterochromatin decondensation, nuclear fusion) and death (apoptosis, necrosis) were observed, especially in heat-shocked specimens. While the number of total and normal nuclei decreased following heat shock, the frequency of apoptosis increased during a short period (7 days) after heat shock. During a 30-day period following heat shock, the frequency of necrosis in fasted but not in fully nourished nymphs increased simultaneously with a decrease in the frequency of apoptosis. This finding suggests that the stress promoted by heat shock, but not that associated with heat shock plus fasting, can be dealt with by the apoptosis program. When considering the forms of cell survival, heterochromatin decondensation was more relevant in fully nourished nymphs, whereas nuclear and cell fusions were more important in fasted specimens. The forms of cell survival and cell death reported here may have protected the organ from damage by the stressing agents. In cells with no induction or accumulation of heat-shock proteins, cell death and the forms of cell survival observed here were the probable consequence.

Changes in nuclear phenotypes following cold shock in Panstrongylus megistus (Burmeister).

The nuclear phenotypes of Malpighian tubule epithelial cells of 5th instar male nymphs of the blood-sucking insect Panstrongylus megistus were studied immediately after a short (1 h) cold shock at 0 degrees C, and 10 and 30 days later. The objective was to compare the responses to a cold shock with those known to occur after hyperthermia in order to provide insight into the cellular effect of cold in this species. Nuclei which usually exhibited a conspicuous Y chromosome chromocenter were the most frequent phenotype in control and treated specimens. Phenotypes in which the heterochromatin was unravelled, or in which there was nuclear fusion or cell death were more abundant in the shocked specimens. Most of the changes detected have also been found in heat-shocked nymphs, except for nuclear fusion which generates giant nuclei and which appeared to be less effective or necessary than that elicited after heat shock. Since other studies showed that a short cold shock does not affect the survival of more than 14% of 5th instar nymphs of P. megistus with domestic habit and can induce tolerance to a prolonged cold shock, heat shock proteins proteins are probably the best candidates for effective protection of the cells and the insects from drastic damage caused by low temperature shocks.

Nuclear phenotype changes after heat shock in Panstrongylus megistus (Burmeister).

The nuclear phenotypes of Malpighian tubule epithelial cells of male nymphs of the blood-sucking insect, Panstrongylus megistus, subjected to short- and long-duration heat shocks at 40oC were analyzed immediately after the shock and 10 and 30 days later. Normal nuclei with a usual heterochromatic body as well as phenotypes indicative of survival (unravelled heterochromatin, giants) and death (apoptosis, necrosis) responses were observed in control and treated specimens. However, all nuclear phenotypes, except the normal ones, were more frequent in shocked specimens. Similarly altered phenotypes have also been reported in Triatoma infestans following heat shock, although at different frequencies. The frequency of the various nuclear phenotypes observed in this study suggests that the forms of cell survival observed were not sufficient or efficient enough to protect all of the Malpighian tubule cells from the deleterious effects of stress. In agreement with studies on P. megistus survival following heat shock, only long-duration shock produced strongly deleterious effects.

Chagas disease in an area of recent occupation in Cochabamba, Bolivia.

INTRODUCTION: A descriptive, entomological and seroepidemiological study on Chagas disease was conducted in a place of recent occupation on the outskirts of Cochabamba, Bolivia: Avaroa/Primer de Mayo (population:3,000), where the socio-economic level is low and no control measures have been made available. METHODS: The immunofluorescent antibody test (IFAT) was used for IgG and IgM anti-Trypanosoma cruzi antibodies in filter paper bloodspot eluates from 128 subjects (73 females, 55 males) selected by systematic sampling. Concerning each subject age, gender, birthplace, occupation, duration of residence and building materials used in their houses were recorded. Vectors were captured both in domestic and peridomestic environments. RESULTS: Seropositive, 12.5% (16/128): females, 15.1% (11/73); males, 9.1% (5/55). Average time of residence: 6.1 years for the whole population sample and 7.4 years for the seropositive subjects. Most houses had adobe walls (76. 7%, n= 30), galvanized iron rooves (86.7%) and earthen floors (53. 4%) 80% of the walls had crevices. One hundred forty seven specimens of Triatoma infestans were captured, of which 104 (70.7%) were domestic, and 1 peridomestic Triatoma sordida. Precipitin host identification: birds, 67.5%; humans, 27.8%; rodents, 11.9%; dogs, 8. 7%; cats, 1.6%. House infestation and density indices were 53.3 and 493.0 respectively. We found 21 (14.3%) specimens of T. infestans infected with trypanosomes, 18 (85.7%) of which in domestic environments. DISCUSSION: The elements for the vector transmission of Chagas disease are present in Avaroa/Primer de Mayo and the ancient custom of keeping guinea pigs indoors adds to the risk of human infection. In neighboring Cochabamba, due to substandard quality control, contaminated blood transfusions are not infrequent, which further aggravates the spread of Chagas disease. Prompt action to check the transmission of this infection, involving additionally the congenital and transfusional modes of acquisition, is required.

Survival and molting incidence after heat and cold shocks in Panstrongylus megistus Burmeister.

Survival and molting incidence were studied after heat (40 degrees C) and cold (0 degree C) shocks in specimens of Panstrongylus megistus with the aim of establishing its response to temperature stress under laboratory rearing conditions and to understand occasional changes in the biological characteristics of specimens captured in nature. The response to the thermal shocks was found to vary as a function of the temperature and duration of the shock, developmental phase and sex of the specimens, and in certain cases, the insect habit and nourishment conditions. P. megistus specimens were found to be less resistant to the heat shock assay than Triatoma infestans, another reduviid species. The short cold shock affected survival of P. megistus more than did the heat shock, survival of fully-nourished specimens being preferential. The response of adults to the short cold shock was affected by sex, males being generally less resistant. The insect sylvatic habit was found to seldom affect the thermal shock response established for specimens with domestic habit. A decrease in molting frequency and sometimes a slowdown of the molting rate were found after the short heat and cold shocks, possibly promoted by change in hormonal balance, and differing from patterns reported for T. infestans. The results indicate that no generalization should be made for different reduviid species in terms of the effects of temperature shocks.

[Triatoma infestans: importance, control and elimination of the species in the state of São Paulo, Brazil]. / Triatoma infestans: importância, controle e eliminação da espécie no Estado de São Paulo, Brasil.

Considerations about the control of the vectors of Chagas' disease are made in the State of Sao Paulo, mainly those activities that led to the elimination of T. infestans. First of all, the authors discuss different aspects of the biology of T. infestans mainly those that permitted it to adapt itself in rural areas of the State in the first middle of the century. Secondary factors that helped the control such as rural exodus are also analysed. The article shows that since 1965 the control became a campaign with different phases due to the epidemiological situation, the acquired knowledge and the entomological surveillance. After 25 years of work, the elimination of all the focus of T. infestans was finally reached and the campaign was ended. However, due to the possibility of reintroduction of the vector in rural areas by passive transportation besides the presence of secondary vectors (T. sordida and P. megistus) in several localities, the vector control activities were not interrupted and the surveillance is continuous.

Triatoma infestans: importância, controle e eliminação da espécie no Estado de São Paulo, Brasil / Triatoma infestans: importance, control and elimination of the species in the state of São Paulo, Brazil

Os autores tecem considerações a respeito do controle dos transmissores da doença de Chagas no Estado de São Paulo, especialmente sobre as atividades que levaram à eliminação do T. infestans. Inicialmente, fazem um apanhado histórico e apresentam as características principais da espécie que possibilitaram sua adaptação à situação vigente na zona rural do Estado, na primeira metade do século. Destacam também os fatores coadjuvantes às ações de controle, particularmente o êxodo rural. Mostram que a partir de 1965 o combate tomou a forma de uma verdadeira campanha, com fases distintas em função das alterações epidemiológicas, experiência adquirida e pressão dos custos. Descrevem cada fase: arrastão, expurgo seletivo, prioridades e vigilância entomológica. Após 25 anos de trabalho a campanha foi considerada encerrada, com a eliminação dos focos da espécie do planalto paulista. Porém, em função da possibilidade da reintrodução de T. infestans (transporte passivo) e da presença, em diversas localidades, de exemplares de espécies vetoras semidomiciliares (T. sordida e P. megistus) as atividades de controle não foram interrompidas e em conseqüência continua em andamento a Vigilância/ Manutenção.

Considerations about the control of the vectors of Chagas' disease are made in the State of Sao Paulo, mainly those activities that led to the elimination of T. infestans. First of all, the authors discuss different aspects of the biology of T. infestans mainly those that permitted it to adapt itself in rural areas of the State in the first middle of the century. Secondary factors that helped the control such as rural exodus are also analysed. The article shows that since 1965 the control became a campaign with different phases due to the epidemiological situation, the acquired knowledge and the entomological surveillance. After 25 years of work, the elimination of all the focus of T. infestans was finally reached and the campaign was ended. However, due to the possibility of reintroduction of the vector in rural areas by passive transportation besides the presence of secondary vectors (T. sordida and P. megistus) in several localities, the vector control activities were not interrupted and the surveillance is continuous.

[Triatoma brasiliensis Neiva, 1911 (Hemiptera, Reduviidae): observations on dynamics of feeding behavior in an artificial ecotope]. / Triatoma brasiliensis (Neiva,1911) (Hemiptera, Reduviidae): observações sobre seu comportamento em relação fonte alimentar em galinheiro experimental.

In order to study the behavior of triatomine bugs under artificial ecotopic conditions, an experimental, double-walled poultry coop was built. Its inside walls were made of sun-dried clay blocks which could be broken apart for better access to the insects hiding in their crevices. The coop was subdivided into two main areas of different sizes using wire netting and a wall made of perforated bricks. The chickens, the only food source available for the triatomines, were kept in the larger, rectangular area, further subdivided diagonally. The triatomines were released in the smaller area and were allowed to migrate to the larger one though the holes in the bricks. Information about migration, survival, and hiding places of fourth-stage nymphs of Triatoma brasiliensis was thus gathered. At 30-day intervals, over a period of six months, the inner walls of the house were taken apart and inspected. The nymphs were marked with different colors according to the month and place of capture. It was found that 88.5% of the nymphs were captured near the roost and found engorged. After having taken their first blood meal, they would remain in the vicinity of their food source. The results obtained are considered epidemiologically relevant, and particularly so as pertaining to the control of the vectors of Chagas disease.

[The prevalence, infection indices and feeding habits of the Triatominae captured in an area of epidemiological surveillance]. / Prevalência, índices de infecção e hábitos alimentares de triatomíneos capturados em uma área de vigilância epidemiológica.

Prevalence of triatomines and their infection by Trypanosoma cruzi, were studied during 1974-1981, in the Administrative Region of Campinas, São Paulo, Brazil. This investigation was continued (1982-1986), blood meal tests with bird, marsupial, rodent and human antisera having been added. Panstrongylus megistus, predominant in the region, whose winged instars are often found infected by T. cruzi in inhabited houses, displayed and alimentary eclecticism. From a sample of 7,785, 14.78 percent reacted positively with human antisera. A similar, but less marked behaviour was observed in Rhodnius neglectus. Triatoma sordida, the second most important species in the Region, was neither demonstrated to be feeding on humans nor to be infected by T. cruzi. Changes in their natural habitat probably caused sharp rises in T. arthurneivai populations in the human dwellings. The capture of three specimens of Microtriatoma borbai marks the first occurrence of this species in the State São Paulo. "Focus investigation" in areas in the surveillance phase can detect triatomines in the process of domiciliation.

[Survival and occurrence of molting in Triatoma infestans Klug (Hemiptera, Reduviidae) after temperature shock]. / Sobrevivência e ocorrência de muda em Triatoma infestans Klug (Hemiptera, Reduviidae) após choque de temperatura.

Survival and molting occurrence were studied in specimens of Triatoma infestans over 30 days after temperature shocks. Hyperthermal and hypothermal shocks could be found to affect both survival and molting incidence as a function of temperature and period of the development phase and sex of the specimens. Considering the various test conditions, the shock at 0 degree C for 12 h was found to elicit the most deleterious effect, whereas shocks at 40 degrees C and 0 degree C even for 1 h are interpreted as affecting the hormonal balance which controls molting. Cases of a rise in post-shock survival are suggested to have been favored by heat-shock protein action.