The viral nature of Toddia França, 1912

Toddia França, 1912 under the light microscope occurs as inclusion corpuscles in the cytoplasm of erythrocytes of cold-blooded vertebrates sometimes accompanied by crystalloid bodies. Its position among the protozoans or the viruses has been discussed by some authors, but remained unclear. To elucidate this problem we studied Toddia from a Brazilian frog (Leptodactylus ocellatus) by electron microscopy. In the cytoplasm of the infected cells we found no protozoan, but rather virus-like particles often hexagonal in outline, averaging 195 nm excluding their two involving membranes, and presenting a central area of variable electron density. Particles at different stages of development were generally found around or on area lighter density than the cytoplasm. which resembled a virus synthesis site. At high magnification, the nuclear or cytoplasmic crystals allied to Toddia resembled the crystalline lattice of the inclusion bodies associated with the polyhedrosis viruses and poxviruses from insects, of the capsules of granulosis viruses and of other protein crystals in ultrathin sections. Cytochemical tests in Toddia corpuscles displayed exclusively the presence of deoxyribonucleic acid. These findings indicate that Toddia is not a protozoan and demonstrate that it is in all probability a viral inclusion corpuscle. Taking into account the nucleic acid type found in its structure (DNA) and the hexagonal shape usually shown in ultrathin sections by its component particles, which have a cytoplasmic site of synthesis and assembly, we tentatively relate Toddia with the so-called "Icosahedral Cytoplasmic Deoxyriboviruses". We believe that the present paper gives the first report of virus-like particles in L. ocellatus.

Studies in search of a suitable experimental insect model for xenodiagnosis of hosts with Chagas' disease. 1 - Comparative xenodiagnosis with nine triatomine species of animals with acute infections by Trypanosoma cruzi

In search of a suitable vector species for xenodiagnosis of humans and animals with chronic Chagas' disease we first investigated the reactions of different vector species to acute infection with Trypanosoma cruzi. Vector species utilized in this study were: Triatoma infestans, Rhodnius prolixus and Triatoma dimidiata, all well adapted to human habitats; Triatoma rubrovaria and Rhodnius neglectus both considered totally wild species; Panstrongylus megistus, Triatoma sordida, Triatoma pseudomaculata and Triatoma brasiliensis, all essentially sylvatic but some with domiciliary tendencies and others restricted to peridomestic biotopes with incipient colonization of human houses after successful eradication of T. infestans. Results summarized in Table IV suggest the following order of infectivity among the 9 studied vector species: P. megistus with 97.8% of infected bugs, T. rubrovaria with 95% of positive bugs a close second followed by T. Pseudomaculata with 94.3% and R. neglectus with 93.8% of infected bugs, almost identical thirds. R. prolixus, T. infestans and T. dimidiata exhibited low infection rates of 53.1%, 51.6% and 38.2% respectively, coupled with sharp decreases occuring with aging of infection (Fig. 1). The situation was intermediate in T. brasiliensis and T. sordida infection rates being 76.9% and 80% respectively. Results also point to the existence of a close correlation between prevalence and intensity of infection in that, species with high infection rates ranging from 93.8% to 97.8% exhibited relatively large proportions of insects (27.3% - 33.5%) harbouring very dense populations of T. cruzi. In species with low infection rates ranging from 38.2% to 53.1% the proportion of bugs demonstrating comparable parasite densities was at most 6%. No differences attributable to blood-meal size or to greater susceptibility of indigenous vector species to parasites of their own geographical area, as suggested in earlier...

Features of reproduction in laboratory-reared Aedes fluviatilis (Lutz, 1904) (Diptera: Culicidae)

Observations were made on 100 female Aedes fluviatilis (Lutz) maintained individually from the first blood meal onwards and allowed one blood meal during each oviposition cycle, 5% dextrose solution been supplied throughout life. The average length of live was 49.9 ± 17.8 days; the logarithm of the mortality rate increased proportionately to physiological age. The insects took an average of 7.3 ± 3.2 blood meals and produced a mean of 7.9 ± 3.7 clutches of eggs. There was a progressive decrease, proportional to advancing physiological age, in the mean numbers of eggs laid in successive oviposition cycles, in the intervals between blood feeding oviposition, and in the numbers of larvae that hatched. Delayed oviposition, transient sterility and a total loss of fertility were also recorded.

Behavior of triatomines (Hemiptera, Reduviidae) vectors of Chagas' disease. II. Influence of feeding, lighting and time of day on the number of mating, mating speed and duration of copulation of Panstrongylus megistus (Burm, 1835) under laboratory conditions

To determine in influence of feeding, lighting and time of day on the copulating behavior of Panstrongylus megistus, 480 insect pairs were divided into four groups of 120 each and tested in the following respective situations: without food deprivation (F.D.), with five days of F.D., with ten days of F.D., and with 20 days of F. D. The tests were performed between 9:00 a.m. to 12:00a.m. and 7:00 p.m. to 10:00 p.m., with light (700-1400 lux) and in the dark (1.4-2.8 lux) and behavior was recorded by the time sampling technique. Mating spped (MS) and duration of copulation (DC) were also calculated for each situation. The maximum frequency of copulation was observed after five days of F.D., at night, in the dark (n = 16), and the minimum was observed for recently-fed pairs, at night, with light (n = 4). Males approached females more often than females approached males. MS was lowest in pairs with twenty days of F.D., at night, with light (X = 23.0 ± 16.0 minutes), and highest in recently-fed pairs, during the day, with light (X = 2.9 ± 2.5 minutes). DC was shortest in recently-fed insects, during the day, in the dark (X = 23.5 ± 6.7 minutes), and longest in recently-fed animals, at night, in the dark (X = 38.3 ± 6.9 minutes).

Behavior of triatomines (Hemiptera: Reduviidae) vectors of Chagas' disease: III. Influence of the number of matings on the fecundity and fertility of Panstrongylus megistus (Burm. 1835) in the laboratory

A study of the effect of mating in the fecundity and fertility of females of P. megistus fed on pigeon blood every 14 days, was carried out in the laboratory. Two groups were constituted: I - females which mated only once; II - females which stayed always with the males. Only 56.7% of group I females laid fertile eggs, while as much as 90% of group II females laid fertile eggs. The duration of the fertile oviposition was greater in the females which stayed always with the males. Some females of this group were able to mate up to seven times throughout their life-span. This fact render useless sterile males in the control of these insects. It is suggested that the components of pigeon's blood used for feeding the triatomines could have an influence upon the fecundity and fertility of the female sof the two groups.

The study of insect blood-feeding behaviour: 2. Recording techniques and the use of flow charts

This paper continues a discussion of approaches and methodologies we have used in our studies of feeding in haematophagous insects. Described are techniques for directly monitoring behaviour: electrical recording of feeding behaviour via resistance changes in the food canal, optical methods for monitoring mouthpart activity, and a computer technique for behavioural event recording. Also described is the use of "flow charts" or "decision diagrams" to model interrelated sequences of behaviours.

Evolution of holometabola insect digestive systems: physiological and biochemical aspects

This review takes into account primarily the work done in our laboratory with insects from the major Holometabola orders. Only the most significant data for each insect will be presented and a proposal on the evolution of Holometabola insect digstive systems will be advanced.

Midgut of Lepidopteran pupae is a major depot of sequestered, mobilizable, ecdysteroids

Extirpation of endocrine organs - a classic maneuver in hormonal research - has been difficult or impossible in the case of the prothoracic glands (PG) of insects. In larval and pupal Lepidoptera the glands are virtually inaccessible unless one sacrifices the insect. Even then, the PG are not easy to remove in their entirety. Consequently, in order to obtain viable preparations lacking PG, one costomarily makes use of abdomens isolated by litigation or surgery.

The Azadirachtins: potent insect growth inhibitors

In the course of their coevolution with insects, plants have learnt to protect themselves by chemical means. Semiochemical act as antifeedants or deterrents, others by disrupting growth and development. By use of the Epilachna varivestis bioassay we isolated from Azadirachta indica seed a group of triterpenoids which interfee with larval growth and development in ppm range. Main components are the azadirachtins A and B with identical biological activity. Various other azadirachtins were obtained, either as minor seed components or by chemical modification of the naturally occuring compounds. Structure vs. activity relation studies enabled us to postulate a basic structural element that should still be biologically active and with much simpler chemical structure than natural compounds. What underlies the biological activity of these insect growth inhibitors? Their interference with the hormonal regulation of development and reproduction has been studied in Locusta migratoria and Rhodnius prolixus. In addition, tritiated dihydroazadirachtin A was used. With this approach, a precise correlation between administered dose, resulting effects, and retention of the compound was established. The azadirachtins either interrupt, delay, or deviate whole developmental programs. Results from these studies provide another chemical probe for studies in insect endocrinology and physiology.

The roles of haemolymphatic lipoproteins in the oogenesis of Rhodnius prolixus

The fates of purified 32P-vitellin and 32P-lipophorin were followed in vitellogenic females of Rhodnius prolixus. While the radioactivity from 32P-vitellin 6 hours after injection was found almost exclusively in the ovary, the radioactivity from injected 32P-lipophorin was found distributed among several organs. In the ovary, the radioactivity from 32P-vitellin was associated with the contents of the yolk granules. 32P-lipophorin delivered a great amount of radioactive phospholipids to the ovary with no accumulation of its protein moiety, as observed after its iodination with 131I. The delivery of phospholipids was inhibited at 0ºC and by the metabolic inhibitors, sodium azide and sodium fluoride. Comparison of the radioactivity incorporation from 32P-lipophorin with that of 14C-inulin suggests that the 32P-phospholipids from lipophorin are not taken up by fluid phase endocytosis. The data presented here are compatible with the concept of lipophorin as a carrier of lipids in insects and provide evidence that lipophorin transports phospholipids as shown previously for other classes of lipids. The utilization by the oocytes of the phospholipids transported by lipophorin is discussed.

From a mono to a digenetic life-cycle: how was the jump for flagellates of the family trypanosomatidae?

It has been found that species of the digenetic genus Trypanosoma as well as species of monogenetic trypanosomatids of insects can grow in the lumen of the scent glands of opossums reproducing the cycle they perform in the intestinal tract of their insect hosts. Based on these findings, speculations are made on the incompletely known cycles of many mammalian trypanosomes and on the evolution of the family Trypanosomatidae.

Morphological evidence by scanning electron microscopy of excretion of metacyclic forms of Trypanosoma cruzi in vector's urine

Comparision by scanning electron microscopy (SEM) of Trypanosoma cruzi flagellates attached to the cuticle of the rectal gland of infected Dipetalogaster maxima nymphs, showed marked differences before amd after feeding. Before feeding numerous metacyclic trypomastigotes were observed among the abundant epimastigotes that formed the carpet of flagellates. On the other hand, in insects that were allowed to urinate for 24 hours after a meal, the metacyclics were scarce,indicating that they had been detached by the urine flow. An asymetric type of cell division, probably originating both an epi-and a trypomastigote, was occasionally observed. The occurrence of swellings at different levels of the flagella of epimastigotes suggests that secondary sites of attachment may be common.

Generos Voriinos de Taquinidos chilenos (Diptera: Tachinidae, Voriini)

Nine genera and 16 species of Chilean Voriini Tachinid flies are reported in this paper, with a key to differentiate them, a list of recorded species, its geographic distibution within the country and known hosts. The examined examples belong and are deposited at the Collection of Insects of the Instituto de Entomología (UMCE), Santiago. Neochaetoplagia pastranai Blanchard, 1963, from Argentina, and Velardemyia ica Valencia, 1972, from Perú, are reported as present in Chile, while Nothovoria praestans n. gen. & sp., is described as a new monotypic genus.

Estudos sobre a resistência ao jejum e aspectos nutricionais de Cavernicola lenti Barrett & Arias, 1985 (Hemiptera, Reduviidae, Triatominae)

In order to widen the present knowledge on the biology of this species, a study on the resistance to starvation was carried out among all nymphal stages and the adult stage (male and female). All evolutive stages were weighed on precision scale in three different nutritional situations: fed, non-fed and death registered after starvation. This procedure has allowed us to calculate the amount of blood taken in each stage and during the whole cycle, the average loss of weight during starvation and its relations with the initial weight. The insects were fed on mice and after eclosion or ecdisis they were isolated for observation of the starving period. Throuhout the whole experiment they were kept in a B. O. D./DOB incubator (28ºC and 90%R.U.). The resistance to starvation of the insects has grown from the first stage on (average of 15.5 days) to the fifth stage (average of 75.64 days); on the adult stage, the resistance period was equal to the third stage with an average of 41.76 for the males and 44.82 for the females. The amount of ingested blood was greater at the fifth stage worth 34.14 mg, corresponding to 2,04 times its initial weight. The average weight loss during the starvation was greater at the adult stage (23.95 mg), corresponding to 61.52% of the total weight.

Fonte alimentar e potencial vetorial de Triatoma vitticeps (Stal, 1859) com relação à doença de Chagas humana no estado do Espírito Santo, Brasil (Hemiptera, Reduviidae)

In paralel with several other epidemiologic and entomologic data of 19 Municipalities of Espírito Santo State, Brazil, the feeding pattern of 222 Triatoma vitticeps is studied through precipitin tests. Very high levels of natural infection with Trypanosoma cruzi are observed in adult insects, in contrast with the abscence or minimum degrees of infection among nymphs and human individuals. The precipitin tests showed the contact of the insects with multiple blood sources, chiefly human and birds, followed by rodents and marsupials. The data suggest that T. vitticeps in spite of being highly antropophilic, become infected by T. cruzi in sylvatic ambient and occasionally invade houses. The species doesn't seem to be - at least until now - a good vector in the domestic cycle of Chagas' disease. Several factors seem to be involved in this conclusion, mainly the low density of the insect in the houses, its hardness to coloniza them, its slowness concerning to suction and defecation and possibly its low susceptibility to different T. cruzi strains.