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As a part of our continuing search for insect and arthropod development regulators from Brazilian restinga vegetation of the Rio de Janeiro State, crude extracts, purified fractions and essential oils were submitted to screening tests seeking for biological activities on the development of the insects Rhodnius prolixus, Dysdercus peruvianus, Oncopeltus fasciatus and Rhipicephalus (Boophilus) microplus. Up to now, 102 secondary metabolites have been detected in the fractions, among them monoterpenes, sesquiterpenes and two triterpenes which were obtained from the species, Eugenia sulcata, Pilocarpus spicatus, Manilkara subsericea, Myrciaria floribunda and Zanthoxylum caribaeum. These secondary plant metabolites are considered of interest for the use of studies related to arthropod endocrinology, vector-parasite interaction system, and population control of vector insect and agricultural pest. The observed biological activities were surprisingly high, involving increased mortality, molting and metamorphosis inhibition, paralysis, corporeal deformities, apparition of permanent nymphs, of adultoids and juvenoids, partial or total inhibition of oviposition and egg hatching. These compounds are now being studied further to determine if they may or may not be useful in controlling insect populations and/or interfere with the life cycle and vector transmission of parasites to animal and human populations.

We evaluated the efficacy of the growth regulator triflumuron (TFM) in inducing mortality and disrupting both oviposition and egg hatching in Rhodnius prolixus adult females. TFM was administered via feeding, topically or by continuous contact with impregnated surfaces. Feeding resulted in mild biological effects compared with topical and impregnated surfaces. One day after treatment, the highest mortality levels were observed with topical surface and 30 days later both topical and impregnated surfaces induced higher mortalities than feeding. Oral treatment inhibited oviposition even at lower doses, and hatching of eggs deposited by treated females was similarly affected by the three delivery modes. Topical treatment of eggs deposited by nontreated females significantly reduced hatching. However, treatment per contact of eggs oviposited by untreated females did not disrupt eclosion. Additionally, oral treatment increased the number of immature oocytes per female, and topical treatment reduced the mean size of oocytes. TFM also affected carcass chitin content, diuresis, and innate immunity of treated insects. These results suggest that TFM acts as a potent growth inhibitor of R. prolixus adult females and has the potential to be used in integrated vector control programs against hematophagous triatomine species.

A chemical investigation and bioassays against fifth-instar nymphae of the hematophagous insect Rhodnius prolixus, vector of Chagas disease, were conducted with the essential oil from Zanthoxylum caribaeum. The main results may be summarized as follows: (i) 54 components were identified, corresponding to 90.4% of the relative composition; sesquiterpenes (47.3%) and monoterpenes (41.2%) are the major constituents; (ii) muurola-4,5-trans-diene and isodaucene are described for the first time as chemical constituents of the essential oil from leaves of this species; (iii) topical treatment with the crude essential oil induced high levels of paralysis (from 18.88 to 33.33%) and mortality (from 80 to 98.9%) depending on the dose applied (0.5 to 5.0 μl per insect); (iv) feeding treatment with the crude essential oil also induced high levels of mortality (from 48.8 to 100%) but low levels of paralysis (from 2.22 to 7.77%) depending on the dose applied (0.5 to 5.0 μl/ml of blood); (v) in the continuous treatment, only the dose of 5.0 μl/cm(2) was able to promote statistical significant levels of mortality (63.3%) but no paralysis were detected. However in this group, occasionally, only few insects displayed malformations of legs and wings after treatment; and (vi) any treatment was able to disrupt the metamorphosis process since the low adult stage emergence observed to all groups was due the high insect mortality. These observations suggest the interference of Z. caribaeum compounds on the triatomine neuroendocrine system. The significance of these results in relation to the relevant biological events in R. prolixus as well as the possible use of insect growth regulators present in Z. caribaeum oil in integrated vector control programs against hematophagous triatomine species is herein discussed.

The effects of azadirachtin A added to the sucrose diet of the adult females on the mortality, oviposition, and hatching of the sand fly vector of American visceral leishmaniasis Lutzomyia longipalpis (Lutz & Neiva, 1912) were investigated. Concentrations of 0.1, 1.0, and 10.0 microg/mg of azadirachtin significantly increased insect mortality in comparison with control insects. The same dose also significantly reduced oviposition but not hatching. After a long development period, significantly fewer adult insects were obtained from eggs hatching by azadirachtin-treated females in a dose-response manner. These results indicate that azadirachtin is a potent sterilizer that could be used against the development of Lu. longipalpis populations and as a tool for studying physiological and biochemical processes in phlebotomine species.

Background: Plants have been recognized as a good source of insecticidal agents, since they are able to produce their own defensives to insect attack. Moreover, there is a growing concern worldwide to develop pesticides with low impact to environment and non-target organisms. Hexane-soluble fraction from ethanolic crude extract from fruits of Manilkara subsericea and its triterpenes were considered active against a cotton pest (Dysdercus peruvianus). Several natural products with insecticidal activity have poor water solubility, including triterpenes, and nanotechnology has emerged as a good alternative to solve this main problem. On this context, the aim of the present study was to develop an insecticidal nanoemulsion containing apolar fraction from fruits of Manilkara subsericea., Results: It was obtained a formulation constituted by 5% of oil (octyldodecyl myristate), 5% of surfactants (sorbitan monooleate/polysorbate 80), 5% of apolar fraction from M. subsericea and 85% of water. Analysis of mean droplet diameter (155.2 ± 3.8 nm) confirmed this formulation as a nanoemulsion. It was able to induce mortality in D. peruvianus. It was observed no effect against acetylcholinesterase or mortality in mice induced by the formulation, suggesting the safety of this nanoemulsion for non-target organisms., Conclusions: The present study suggests that the obtained O/A nanoemulsion may be useful to enhance water solubility of poor water soluble natural products with insecticidal activity, including the hexane-soluble fraction from ethanolic crude extract from fruits of Manilkara subsericea.

Background: Studies were carried out to evaluate the effects of Manilkara subsericea extracts and triterpenes on the development of two species of agricultural pest insects, Oncopelus fasciatus and Dysdercus peruvianus., Results: All treatments of insects with M. subsericea extracts induced mortality, delayed development and inhibited moulting. Some extracts assayed (FH, FB and FD in D. peruvianus, and FH, FB and FEA in O. fasciatus) also produced body deformities in the few adults that emerged. Other extracts (FH, FEA, FB, FD and LET in both insects), however, induced either permanent (overaged) or extranumerary nymphs, both of which were unable to achieve the adult stage and reproductive status. The insects were also treated with triterpenes (α- and β-amyrin acetates) which showed high lethality at 30 days after treatment and delayed the intermoult period so that overaged nymphs were detected., Conclusion: The results indicate that extracts and triterpenes from Manilkara subsericea act as potent growth inhibitors of phytophagous hemipteran nymphs. It is also concluded that the mixture of several different molecules in the extracts used produces synergic effects that do not occur after using the triterpenes (PFT) alone. These secondary metabolites in the M. subsericea extracts can potentially be used in integrated control programmes against crop pests., (Copyright © 2012 Society of Chemical Industry.)

The effects of azadirachtin and ecdysone on the Trypanosoma cruzi population in the Rhodnius prolixus gut were investigated. T. cruzi were rarely found in the gut compartments of azadirachtin-treated larvae. High parasite numbers were observed in the stomach of the control and ecdysone groups until 10 days after treatment and in the small intestine and rectum until 25 days after treatment. High percentages of round forms developed in the stomachs of all groups, whereas azadirachtin blocked the development of protozoan intermediate forms. This effect was counteracted by ecdysone therapy. In the small intestine and rectum, epimastigotes predominated for all groups, but more of their intermediates developed in the control and ecdysone groups. Azadirachtin supported the development of round forms and their intermediates into trypomastigotes. In the rectum, trypomastigotes did not develop in the azadirachtin group and developed much later after ecdysone therapy. The parallel between the effects of azadirachtin and ecdysone on the host and parasite development is discussed on the basis of the present results because ecdysone appears to act directly or indirectly in determining the synchronic development of T. cruzi forms from round to epimastigotes, but not metacyclic trypomastigotes, in the invertebrate vector., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Studies were carried out to evaluate the efficacy of the growth regulator, triflumuron (TFM) (Starycide sc 480 Bayer), for disrupting the development of Rhodnius prolixus fifth-instar nymph by oral, topical or continuous treatment. All treatments were able to induce high levels of mortality, delay development and molt inhibition. Oral treatment induced molt inhibition in all insects that survived at doses of 0.25, 0.50 and 5.0 mg/mL of a blood meal. The highest levels of both mortality in 24 h and molt inhibition were always observed after topical treatment. The lowest doses needed to obtain considerable biological effects were always observed after continuous treatment. In this way, the highest levels of mortality within 30 days were detected after continuous treatment, which also induced an extended inter-molting period, a lower number of over-aged nymphs and the highest level of molting in nymphs that survived. Moreover, the effects of TFM on insects were often displayed in a dose response manner. These results indicate that TFM acts as a potent growth inhibitor of R. prolixus nymphs and has the potential to be used in integrated vector control programs against hematophagous triatomine species.

The effects of azadirachtin A added to the standard diet on the development, mortality, and metamorphosis of Lutzomyia longipalpis Lutz & Neiva, 1912 were studied. Concentrations of 0.1, 1.0, and 10.0 microg of azadirachtin/mg of diet significantly increased larval mortality in comparison with nontreated insects. Concentrations 0.1 and 1.0 microg blocked the molt of larvae, which remained as third instars until the end of the experiment. The 10 microg/mg concentration resulted in greater molt inhibition. In this group, all insects stopped their development as second instars. Simultaneous addition of ecdysone (1 microg/mg) to the standard diet containing azadirachtin counteracted the effects of azadirachtin on mortality and inhibition of ecdysis. These results indicate that azadirachtin is a potent growth inhibitor of L. longipalpis.

Studies on the effects of decapitation, head transplantation, azadirachtin, and ecdysone therapy on the ultrastructural organization of the midgut of Rhodnius prolixus, a vector of the protozoan Trypanosoma cruzi, show a distinct effect on the organization of the epithelial cells. When insects are decapitated or treated with azadirachtin, the ultrastructural organiza tion of these compartments changed significantly and drastically blocked the development of T. cruzi infection. In converse experiments, head transplantation or oral therapy with ecdysone significantly re versed the T. cruzi infectivity and reestablished the organization of the stomach and intestine in decapitated or azadirachtin-treated insects. These results indicat that a brain factor, possibly the prothoracicotropic hormone which stimulates ecdysteroid production on the prothoracic glands, may act directly or indirectly on both the midgut cell organiza tion and the intestinal microenvironment, interfering in the trypanosome survival and infection of the vector R. prolixus., (Copyright 1999 Academic Press.)

Studies were carried out on the effects of azadirachtin on the course of Trypanosoma cruzi infection in the gut of different triatomine vector species. In Rhodinus prolixus the development of T. cruzi clone Dm28c decreased in a dose-dependent manner, and the ED50 of this inhibitory effect was 0.25 microgram azadirachtin/ml bloodmeal. Using this insect, we conducted a long-term experiment which showed that azadirachtin (1.0 microgram/ml bloodmeal) completely blocks the development of T. cruzi even 120 days after treatment with the drug, and after four infectant meals. Similarly, the elimination of T. cruzi in feces and urine was also completely blocked over a period of 50 days after infection in insects treated with azadirachtin. Fifth-instar larvae of R. prolixus, Triatoma infestans, and Dipetalogaster maximus, infected with different clone/strains of T. cruzi, displayed drastic inhibition of trypanosome development when treated with azadirachtin. The discussion of these results focuses on the possibility that azadirachtin may act directly on gut physiology and/or indirectly through the neurosecretory system.