Vitellogenin content in fat body and ovary homogenates of workers and queens of Melipona quadrifasciata anthidioides during vitellogenesis

Vitellogenin (Vg) is an egg yolk protein that is produced primarily in the fat body of most female insects. In the advanced social structure of eusocial honeybees, the presence of the queen inhibits egg maturation in the workers ovaries. However in the stingless bee Melipona quadrifasciata, the workers always develop ovaries and lay a certain amount of eggs while provisioning the brood cells with larval food during what is known as the worker nurse phase. The present work is a comparative study of the presence of Vg in homogenates of the fat bodies and ovaries of the nurse workers, and the virgin and physogastric queens of M. quadrifasciata. The presence of Vg was determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis and immunoblotting using Apis mellifera anti-egg antibody. Vg was not detected in the fat bodies or ovaries of the workers, but it was found in the ovaries of virgin and physogastric queens and in the fat body of physogastric queens. The results are discussed, taking into account the reproductive state of the individuals and the other possible roles of Vg, such as a storage protein for metoabolism of other organs.

The insect neuropeptide prothoracicotropic hormone is released with a daily rhythm: re-evaluation of its role in development.

Prothoracicotropic hormone (PTTH) is the central cerebral neurohormone in insect development. Its release has been believed for decades to be confined to one (or two) critical moments early in each developmental stage at which time it triggers prolonged activation of the prothoracic glands to synthesize and release the steroid molting hormones (ecdysteroids), which elicit developmental responses in target tissues. We used an in vitro assay for PTTH released from excised brains of the bug Rhodnius prolixus and report that release of PTTH does occur at the expected time on day 6, but that this release is merely the first in a daily rhythm of release that continues throughout most of the 21 days of larval-adult development. This finding, together with reports of circadian control of ecdysteroid synthesis and titer throughout this time, raises significant challenges to several features of the current understanding of the hormonal control of insect development. New questions are raised concerning the function(s) of PTTH, its relationship with the prothoracic glands, and the significance of circadian rhythmicity throughout this endocrine axis. The significance of the reported observations derives from the set of entirely new questions they raise concerning the regulation of insect development.

Feeding sources and trypanosome infection index of Rhodnius pallescens in a Chagas disease endemic area of Amador County, Panama

The sylvatic triatomine Rhodnius pallescens is considered to be the most important and widespread vector of Trypanosoma cruzi and Trypanosoma rangeli in Panama. However, its behavior and biological characteristics have only been partially investigated. Thus, to achieve sustainable and efficient control over Chagas disease in Panama, a better understanding of the ecology and biology of R. pallescens is essential. In this study we evaluated R. pallescens host feeding sources using a dot-blot assay, and the trypanosome infection index by PCR analysis in a Chagas disease endemic area of central Panama. It was found that in peridomestic palm trees, 20.3% of the examined bugs had fed on opossums (Didelphis marsupialis). However, we observed an increased anthropophagy (25.4%) for those bugs collected inside houses. Considering the domestic and peridomestic habitats as a whole, the proportion of collected R. pallescens infected with trypanosomes was 87.4%. In the two habitats the predominant infection was with T. cruzi (80-90%). Between 47-51% of the analyzed triatomines were infected with T. rangeli. Mixed infections (40-51%) were also detected. These findings provide a better basis for the implementation of a rational control and surveillance program for Chagas disease in regions where R. pallescens is endemic.

First report of Rhodnius montenegrensis (Hemiptera: Reduviidae: Triatominae) infection by Trypanosoma rangeli

Introduction: This study reports for the first time the infection of Rhodnius montenegrensis by Trypanosoma rangeli. Methods: The triatomines were manually collected in Attalea speciosa in the municipality of Buritis, Rondônia. The identification of the trypanosomatid species was confirmed by multiplex PCR. Results: All of the collected triatomines were R. montenegrensis. The analysis confirmed that all of the adults were infected with the epimastigote form of T. rangeli. Conclusions: This report of a new vector of T. rangeli raises a warning for the State of Rondônia because the simultaneous presence of T. rangeli with T. cruzi in the same geographic region enables the occurrence of mixed infections in hosts and vectors, which complicates the differential diagnosis.

The Triatomines of Northern Peru, with Emphasis on the Ecology and Infection by Trypanosomes of Rhodnius ecuadoriensis (Triatominae)

Information on the distribution and synanthropic behaviour of triatomines is essential for Chagas disease vector control. This work summarises such information from northern Peru, and presents new data on Rhodnius ecuadoriensis - an important local vector infesting 10-35% of dwellings in some zones. Three species are strongly synanthropic and may be suitable targets for chemical control of domestic/peridomestic bug populations. Panstrongylus herreri, the main domestic vector in the area, is probably present in sylvatic ecotopes in the Marañón river system. R. ecuadoriensis and Triatoma dimidiata seem exclusively domestic; biogeographical and ecological data suggest they might have spread in association with humans in northern Peru. Confirmation of this hypothesis would result in a local eradication strategy being recommended. Presence of trypanosome natural infection was assessed in 257 R. ecuadoriensis; Trypanosoma rangeli was detected in 4% of bugs. Six further triatomine species are potential disease vectors in the region (T. carrioni, P. chinai, P. rufotuberculatus, P. geniculatus, R. pictipes, and R. robustus), whilst Eratyrus mucronatus, E. cuspidatus, Cavernicola pilosa, Hermanlentia matsunoi, and Belminus peruvianus have little or no epidemiological significance. A strong community-based entomological surveillance system and collaboration with Ecuadorian public health authorities and researchers are recommended.

Polymorphism of the ITS-2 region of the ribosomal DNA of the Triatominae Rhodnius domesticus, R-pictipes, R-prolixus and R-stali

The polymerase chain reaction-restriction fragment length polymorphism technique (PCR-RFLP) was used to compare Rhodnius domesticus (Neiva & Pinto), R. pictipes (Stal), R. prolixus (Stal) and R. stali (Lent; Jurberg & Galvao) (Hemiptera: Reduviidae). The enzyme BstUI differentiated R. donzesticus, R. pictipes and R. prolixus, and HhaI differentiated R. domesticus, R. pictipes and R. stali. With the fingerprinting analysis generated by these two enzymes, it was possible to clearly identify all four species in the study.

Estudo da fauna de triatomíneos e da ocorrência de doença de Chagas em Monte Negro, Rondônia, Brasil

A doença de Chagas tem como agente etiológico o Trypanossoma cruzi, um protozoário flagelado que pode ser encontrado numa grande variedade de mamíferos e triatomíneos. O Estado de Rondônia, localizado na Amazônia Ocidental, possui um meio ambiente constantemente modificado pelas ações transformadoras do ser humano, resultando em um desequilíbrio, que pode facilitar a transmissão de inúmeros patógenos. Uma grande variedade e quantidade de palmáceas, em especial o babaçu, bem como mamíferos e triatomíneos, podem ser encontrados neste complexo ecossistema. Nesta pesquisa, a fauna de triatomíneos foi identificada em 225 babaçus e por meio de capturas peri e intradomiciliares. Foi realizado, concomitantemente, estudo de soroprevalência para doença de Chagas e a identificação da presença de T.cruzi no trato digestivo dos triatomíneos. Positividade ao T. cruzi foi verificada em 23,7% dos 652 triatomíneos coletados nos babaçus. Estes triatomíneos pertenciam ao gênero Rhodnius e foram classificados em 4 espécies: R. robustus, R. prolixus, R. pictipes e R. milesi. Nas capturas intradomiciliares, dez espécimes do Rhodnius robustus e uma de Panstrongylus geniculattus foram encontrados, sendo que 3% da população foi positiva para doença de Chagas. Na área pesquisada, há potencial de transmissão da doença de Chagas na forma endêmica devido a grande quantidade de triatomíneos, bem como alta freqüência de infecção destes triatomíneos, porém no momento deste estudo não se evidenciou a ocorrência da transmissão.

Aspectos morfológicos de ovos de triatominae: II - Características macroscópicas e exocoriais de dez espécies do gênero Rhodnius Stal, 1859 (Hemiptera - Reduviidae)

Foi realizado estudo morfológico do exocório de ovos de dez espécies de Rhodnius (Hemiptera-Reduviidae) através de microscopia eletrônica de varredura e microscopia óptica, com vistas à classificação taxionômica: Rhodnius domesticus Neiva & Pinto, 1923; R. ecuadoriensis Lent & León, 1958; R. nasutus Stal, 1859; R. neglectus Lent, 1954; R. neivai Lent, 1953; R. pallescens Barber, 1932; R. paraensis Sherlock, Guitton & Miles, 1977; R. pictipes Stal, 1972; R. prolixus Stal, 1859; e R. robustus, 1927. São apresentados dados sobre aspectos gerais das cascas e dos ovos quanto à forma, coloração e tamanho, bem como as modificações que poderão ocorrer quanto à coloração durante o desenvolvimento embrionário pós-oviposição. São elaboradas chaves dicotômicas para a classificação destas espécies, baseadas na utilização de caracteres visíveis pela microscopia óptica e microscopia eletrônica de varredura.

Studies in search of a suitable experimental insect model for xenodiagnosis of hosts with Chagas' disease: 4 - The reflection of parasite stock in the responsiveness of different vector species to chronic infection with different Trypanosoma cruzi stocks

Previous studies (1982,1987) have emphasized the superiority of sylvatic vector species over domestic species as xenodiagnostic agents in testing hosts with acute or chronic infections by T. cruzi "Y" stock. The present study, which is unique in that it contains data on both infectivity rates produced by the same stock in 11 different vector species and also the reaction of the same vector species to seven different parasite stocks, establishes the general validity of linking efficiency of xenodiagnosis to the biotope of its agent. For example, infectivity rates produced by "São Felipe" stock varied from 82.5% to 98.3% in sylvatic vectors but decreased to 42.5% to 71.3% in domestic species. "Colombiana" stock produced in the same sylvatic vectors infectivity rates ranging from 12.5% to 45%. These shrank to 5%-22.5% in domestic bugs. The functional role of the biotope in the vector-parasite interaction has not been eluddated. But since this phenomenon has been observed to be stable and easy to reproduce, it leads us to believe that the results obtained are valid. Data presented also provide increasing evidence that the infectivity rates exhibited by bugs from xenodiagnosis in chronic hosts, are parasite stock specific. For example, infectivity rates produced by "Berenice", "Y", "FL" and "CL" varied in R. neglectus from 26.3% to 75%; in P. megistus from 56.3% to 83.8%; in T. sordida from 28.8% to 58.8% in T. pseudomaculata from 41.3% to 66.3% and in T. rubrovaria from 48.8% to 85%. Data from xenodiagnosis in the same hosts, carrying acute infections by the same parasite stocks, gave the five sylvatic vectors a positive rating of approximately 100%, thus suggesting that the heavy loads of parasites circulating in the acute hosts obscured the characteristic interspecific differences for the parasite stock. Nonetheless these latter were revealed in the same hosts with chronic infections stimulated by very low numbers of the same parasite stocks. Certain observations here described lead us to speculate as to the possibility of further results from other parasite stocks, allowing the association of the infectivity rates produced in bugs by different parasite stocks with the isoenzymic patterns revealed by these stocks.

Domestic Rhodnius ecuadoriensis (Hemiptera, Reduviidae) infestation in Northern Peru: a comparative trial of detection methods during a six-month follow-up

Two passive methods in the assessment of intradomiciliary infestation by Rhodnius ecuadoriensis were tested: (i) the Gomes Nuñez sensor box (GN), (ii) sheets of white typing paper and (iii) one active timed manual method. The study was carried out in the Alto Chicama River Valley, Province of Gran Chimú, Department of La Libertad. The study design consisted of an initial searching of triatomines inside of the domestic environment by the manual capture active procedure (man/hour) covering all the studied houses. Then, matched pairs of GN boxes and paper sheets were simultaneously installed in the bedrooms of 207 households distributed in 19 localities. A comparative prospective trial of these passive detection devices were monitored at 2, 4 and, finally 6 months follow-up. Parasitological Trypanosoma rangeli and/or T. cruzi infections were investigated in two houses with high level of infestation by R. ecuadoriensis. 16.9% of the 207 households investigated by an initial active manual method were infested with R. ecuadoriensis. The proportion of infested houses fluctuated from 6.2 to 55.5% amongst the 19 localities investigated. T. rangeli natural infection was detected in R. ecuadoriensis specimens collected in two households. Parasite rates in the bugs ranged from 16.6 to 21.7% respectively. The most striking fact was an average rate of salivary gland infection ranging from 7.4 to 8.3%. At the end of the sixth month period, a cumulative incidence of 31.4% of positive GN boxes against 15.9% for paper sheets was recorded. All three methods combined detected domestic infestation in 129 (62.3%) of the 207 houses studied in the 19 localities. The range of houses infested varies from 6.7% to 92.9%. In areas with low bug density infestation rates, the methodology experienced in our studies, seems to be the best choice for investigations on domestic R. ecuadoriensis populations.

Eco-epidemiological aspects of Trypanosoma cruzi, Trypanosoma rangeli and their vector (Rhodnius pallescens) in Panama

The eco-epidemiology of T. cruzi infection was investigated in the Eastern border of the Panama Canal in Central Panama. Between 1999 and 2000, 1110 triatomines were collected: 1050 triatomines (94.6%) from palm trees, 27 (2.4%) from periurban habitats and 33 (3.0%) inside houses. All specimens were identified as R. pallescens. There was no evidence of vector domiciliation. Salivary glands from 380 R. pallescens revealed a trypanosome natural infection rate of 7.6%, while rectal ampoule content from 373 triatomines was 45%. Isoenzyme profiles on isolated trypanosomes demonstrated that 85.4% (n = 88) were T. cruzi and 14.6% (n = 15) were T. rangeli. Blood meal analysis from 829 R. pallescens demonstrated a zoophilic vector behavior, with opossums as the preferential blood source. Seroprevalence in human samples from both study sites was less than 2%. Our results demonstrate that T. cruzi survives in the area in balanced association with R. pallescens, and with several different species of mammals in their natural niches. However, the area is an imminent risk of infection for its population, consequently it is important to implement a community educational program regarding disease knowledge and control measures.

COLONIZATION OF PALM TREES BY Rhodnius neglectus AND HOUSEHOLD AND INVASION IN AN URBAN AREA, ARAÇATUBA, SÃO PAULO STATE, BRAZIL

The objective of this study is to report on the colonization of palm trees by Rhodnius neglectus, its invasion in an urban area, in Araçatuba - São Paulo, and the control and surveillance measures that have been put in place. Domiciliary triatomine searches occurred in apartments upon the inhabitants' notification. The collected insects were identified and examined for natural infection and food sources with a precipitin test. To search the palm trees, tarps were used to cover the floor, and a “Munck” truck equipped with a tree-pruning device was utilized. Chemical control was performed with the utilization of a manual compression. In 2009, 81 specimens of Rhodnius neglectus were collected from the domiciles by the population. The precipitin test revealed a presence of human blood in 2.7% of the samples. Entomological studies were carried out in these domiciles and in those located within a radius of 200 meters. The search performed in the palm trees resulted in the capture of 882 specimens of triatomines, negative for tripanosomatids. Mechanical and chemical controls were carried out. New searches conducted in the palm trees in the same year resulted in the capture of six specimens. The mechanical and chemical controls of the palm trees, together with the population's work, proved to be effective, therefore preventing these insects' colonization of the city's domiciles.

Synanthropic triatomines (Hemiptera, Reduviidae) in the state of Pernambuco, Brazil: geographical distribution and natural Trypanosoma infection rates between 2006 and 2007

INTRODUCTION: The present study shows a descriptive analysis of triatomine occurrence and its natural Trypanosoma infection rates in the state of Pernambuco, Brazil, between 2006 and 2007. METHODS: Entomological data for the species, such as specimens captured in both intra and peridomiciles and natural infection index, were obtained via domiciliary capture in 147 municipalities from 11 Regional Managements of Health. The database was obtained from a sample of insects (100% infected and 20% non-infected) sent to the Central Laboratory of Pernambuco. RESULTS: A total of 18,029 triatomines were analyzed from 138 municipalities of the state. Triatoma pseudomaculata (35%), Triatoma brasiliensis (34%), and Panstrongylus lutzi (25%) were the most captured species. These species also showed a widespread geographical distribution in the state. Panstrongylus megistus, Triatoma petrocchiae, Triatoma melanocephala, Triatoma sordida, Rhodnius nasutus, Rhodnius neglectus, and Triatoma infestans showed more limited geographical distribution and lower relative abundance. The parasitological research showed that 8.8% of the triatomines were naturally infected with flagellates morphologically similar to Trypanosoma cruzi and 91.3% of them were captured inside houses in 113 municipalities. P. lutzi showed the highest rates of natural infection. CONCLUSIONS: After the control of T. infestans, synanthropic species, such as T. brasiliensis, T. pseudomaculata, and P. lutzi, maintain the risk of T. cruzi transmission to humans in the state of Pernambuco. These species are widely distributed, and infected specimens have been found inside houses. Thus, an enhanced surveillance and vector control of Chagas disease is recommended in Pernambuco.

Characteristics of Triatomine infestation and natural Trypanosoma cruzi infection in the State of Rio Grande do Norte, Brazil

Abstract: INTRODUCTION Natural and artificial ecotope infestation by the kissing bug triatomines and their colonization and infection by Trypanosoma cruzi , the Chagas disease agent, were evaluated in nine municipalities of the State of Rio Grande do Norte, Brazil. METHODS Following identification, triatomine intestinal contents were analyzed by direct microscopic examination, xenoculture, and polymerase chain reaction (PCR) for parasite detection. Trypanosoma cruzi isolates were genotyped using three different markers. RESULTS Of 842 triatomines captured, 65% were Triatoma brasiliensis , 17.8% Triatoma pseudomaculata , 12.5% Panstrongylus lutzi , and 4.7% Rhodnius nasutus . Triatoma brasiliensis and P. lutzi adults were found in the intradomicile. T. brasiliensis, T. pseudomaculata , and R. nasutus nymphs and adults were found in the peridomicile and wild environment. Intradomiciliary and peridomiciliary infestation indexes were 5.6% and 33.7%, respectively. In the peridomicile, chicken coops were the most infested ecotope. The T. cruzi triatomine infection rate was 30.2%, of which PCR detected 29%. P . lutzi (78.1%), T . brasiliensis (24.5%), and T . pseudomaculata (22.7%) were the most infected species. TcII and III genotypes were detected in T. brasiliensis and TcIII in P. lutzi . CONCLUSIONS T. brasiliensis was found in all environments and most ecotopes with high T. cruzi infection rates. High infection rates were also detected in T . pseudomaculata and P. lutzi , suggesting their role in the interchange between the wild and peridomestic transmission cycles. The combination of PCR, microscopic examination, and xenoculture contributed to improving T. cruzi infection evaluation in triatomine bugs. The TcII and TcIII genotypes were predominant in the study area.

Estudos anatômicos e histológicos sôbre a subfamília Triatominae (Heteroptera, Reduviidae): Parte XXI. Estudo comparado do sistema traqueal em Triatoma, Panstrongylus e Rhodnius

É feito um estudo comparado do sistema traqueal nos Triatomíneos. Notou-se que os ramos traqueais secundários apresentam diversidades quanto quanto ao número e à disposição topográfica das traquéias, inclusive quando se tratando de um mesmo exemplar. Neste trabalho é descrito o método de infiltração de corantes, segundo GAEBLE, e modificado por nós quando adaptado a uma bomba de vácuo. Os troncos traqueais principais permanecem inalteráveis, seguindo uma disposição comum e já descrita em Triatoma infestans. Porém, devido as modificações encontradas nos ramos secundários acreditamos que êste sistema não devea ser considerado para, juntamente com outros caracteres, diferenciar o indivíduo. O estudo anatômico do sistema traqueal nos gêneros Triatoma (T. vitticeps, T. maculata, T. brasiliensis, T. sordida), Panstrongylus (P. megistus) e Rhodnius (R. prolixus) é sempre correlacionado com o sistema traqueal de Triatoma infestans. É aproveitada a ocasião para se observar com maiores detalhes a disposição das traquéias em alguns órgãos. Em Triatoma vitticeps notou-se o sistema traqueal em relação aos tubos de Malpighi; em Triatoma brasiliensis o sistema em relação as gônadas e, finalmente, em Panstrongylus megistus as traquéias em relação ao aparelho digestivo, massa ganglionar nervosa situada no tórax e, também, gônadas femininas.