Sôbre alguns Phlebotomus e Brumptomyia da Colômbia (Diptera, Psychodidae)

The first case of Kala-azar in Colombia was discovered in Soledad, S. Vicente do Chucuri, Dept. Santander, by Gast-Galvis who viscerotomized a three year old girl deceased in December, 1943. In 1944, fifty-three Phlebotominae were collected in the chicken pen of the girl's house, two new species included. Mangabeira helped by A. Gast Galvis, Juan Antonio Montoya and E. Osorno Mesa, collected some Phlebotomus in that country. The geographical distribution of the species of Phlebotomus collected in Colombia (P. abonnenci, P. camposi, P. columbianus, P. dubitans, P. gasti, P. montoyai, P. saulensis, P. serranus, P. triramulus) and two species of Brumptomyia (B. beaupertuyi and b mesari), are included. our description of the male P. columbianus is based on some specimens found in association with females. However, doubts exist about such association of sexes. There is no correspondence between the length of the spicules and the ducts of spermathecae. Besides, the specimens were not obtained by raising. The following new species are described and compared with previously known ones: a) Phlebotomus gasti sp. n. differs from the other species by a protruding tubercle in the gubernaculum. It has also fewer setae in the tuft of the basistyle, a different length of the inferior gonapophyses, and a differently shaped clasper. b) Phlebotomus dubitans sp. n. differs from P. walkeri and P. deanei (according to personal information from O. Theodor, who examined the types, they are identical to P. williamsi and P. sericeus respectively), mainly because these species have the inferior gonapophyses larger than the basistyle and fewer setae in the basistyle. P. evandroi is separated by the shape of the claspers and by the tuft of setae of the basistyle. P. marajoensis is the closest relative to P. dubitans. There is a possibility of their being synonymous. On the other hand, they can be differentiated by the existence of three extra distal spines in P. marajoensis. There is also a difference in their palpal indexes: for marajoensis I - II - IV - III - V, and for dubitans I - IV (III - II) - V. We notice, too, that the inferior gonapophyses in P. marajoensis is a little shorter. P. marajoensis has a long seta in the basistyle (clearly shown in the original drawing), not found in the new species. c) Phlebotomus montoyai sp. n.: The closest relatives are P. noguchii, P. peruensis, P. pescei, P. quinquifer and P. rickardi. They differ from the new species by the number and length of the setae of the basistyle tuft which are more numerous and longer in the new species. The shapes of their claspers are also different. Other differences are: the basal portion of the basistyle in P. noguchii is very wide (in montoyai it is narrower); the intermediate spine of the dististyle is located on a protruding tubercle ( in the new species there is hardly a tubercle); the spicules are long, and the inferior gonapophyses is longer than the basistyle. P. quinquifer and P. rickardi have a shorter dististyle and narrower wings, with different venation. The main difference, however lies, in the M4, which ends almost at the level of the junction of M1 with M2 (in P. montoyai the M4 ends far behind). In P. peruensis and P. pescei the intermediary spine of the dististyle is closer to the distal spine than to the basal one, whereas in the new species it is situated between the two pairs. Their inferior gonapophyses is longer than the basistyle. d) Brumptomyia mesai sp. n. - Closest relatives are: B. hamatus, B. pentacanthus, B. beaupertuyi which are easily separated from the new species because the tufts of their basistyle have thin and differently shaped hairs. Also their claspers are shaped differently. B. avellari is also easily recognized on account of the twisted aspect of its clasper and because the basal tuft of the basistyle has few setae, B. brumpti tuft of setae arise directly from the basistyle; these setae are stronger than those of the new species. It has 8 blade-like setae located on the inner surface of the distal half, whereas the new species has only six setae. In B. brumpti, there are three median and two terminal spines in the dististyle; in the new species, there are two median and two terminal spines and one between them, which is closer to the two median spines. The comparison with B. galindoi is based in a specimen determined by Fairchild and deposited in the entomological collection of the "Faculdade de Higiene e Saúde Pública da Universidade de S. Paulo". The genitalia of the new species is much shorter, in galindoi the inferior gonapophyses is 0,8 mm long whereas in B. mesai it hardly reaches 0,6 mm. The shape of the clasper and the distribution of its setae are different. The sub-median lamellae, besides being longer in B. galindoi are also longer in comparison with the other parts of the genitalia. The gubernaculum of the new species is longer, thinner, and more pointed; in B. galindoi it is shorter and triangular. In the drawing published by Fairchild and Hertig 91947), the basistyle shows 8 blade-like setae on the distal half, whereas in the new species only six are found.

Pneumocystis carinii: its incidence in rodents and enhancement of the infection by corticosteroids

A survey was made on the incidence of Pneumocystis carinii in 361 rodents including sewer rats, albino rats, albino mice, guinea-pigs and rabbits. P. carinii was found in 4 of the 215 Rattus norvegicus examined (1,8%). These results accord with recent observations but disagree with investigations made by the researchers who first studied this parasite in the past when high indexes of infection were found. However, in 20 albino rats treated with corticosteroids (betamethazone) we found 8 positive (40%) and in 20 albino mice treated by the same way, 9 were positive for P. carinii (45%). These results confirm the opportunistic character of P. carinii in rodents already well demonstrated in man.

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.

Studies on Trypanosoma rangeli Tejera 1920: VI. Developmental pattern in the haemolymph of Rhodnius prolixus

The morphological sequence of Trypanosoma rangeli development in the body cavity of Rhodnius prolixus is described. The metacyclic trypanosome is the product of successive division and transformation during the intra and extracellular development in the haemocoele. The significance of the early invasion of T. rangeli into the haemolymph is discussed. The epidemiological importance of the developmental pattern of T. rangeli in the vectors haemolymph and the host-response to the parasite are considered.

Differential effect of culture epimastigotes and blood-form Trypamastigotes on normal mouse splenocyte responsiveness to mitogens

Blood form trypomastigotes of the Y strain of T. cruzi, produced a strong inhibition of the blastogenic response to T and B cell mitogens, of the C3H/He, C57BLand BALB/cJ strains of mice, while culture epimastigotes of the Y strain kept in a medium that allows parasite growth at 26°. 30° and 37°C produced a strong stimulatory effect that was even higher than the effect of the mitogens alone. Both the inhibitory or the stimulatory effects were dose-dependent. The stimulatory effect of epimastigotes was also temperature-dependent producing increasedstimulation indexes as the temperature of parasite cultures was raised. Metabolically active,living parasites seemed to be necessary for an improved lymphocyte stimulation suggesting a potential role of secreted metabolites as polyclonal activators of mouse lymphocytes.

The cell biology of mosquito vitellogenesis

Insect vitellogenesis involves coordinated activities of the fat body and oocytes. We have studied these activities at the cellular level in the mosquito. During each vitellogenic cycle, the fat body undergoes three successive stages: 1) proliferation of biosynthetic organelles, 2) vitellogenin synthesis, 3) termination of vitellogenin synthesis and degradation of biosynthetic organelles by lysosomes. Analysis with monoclonal antibodies and radiolabelling demonstrated that the mosquito yolk protein consists of two subunits (200-kDa and 65-kDa). Both subunits are glycosylated, their carbohydrate moieties are composed of high-mannose oligosaccharides. The yolk protein subunits are derived from a single 220 kDa precursor detected by an in vitro translation. Oocytes become competent to internalize proteins as a result of juvenile hormone-mediated biogenesis of endocytotic organelles. The yolk protein is then accumulated by receptor-mediated endocytosis. A pathway of the yold protein and factors determining its routing in the oocyte have been studied.

Atypical rotavirus among diarrhoeic children living in Belém, Brazil

Atypical rotaviruses were detected in faeces from two diarrhoeic children living in Belém, Pará, Brazil. Rotavirus particles were detected by electron microscopy and the RNA electrophoresis showed patterns which were compatible with group C rotaviruses. Tests for the presence of group A antigen by enzyme-linked-immunosorbent assay (ELISA) were negative. The two children had three successive rotavirus infection and in both cases the atypical strains were excreted at the time of the third infection, causing a mild and short-lasting disease.

Stability of isoenzyme and kinetoplast DNA (k-DNA) patterns in successively cloned Trypanosoma cruzi populations

Four Trypanosoma cruzi strains from zymodermes A, B, C and D were successively clonedon BHI-LIT-agar-blood BLAB). Twenty clones from the first generation (F1), 10 from The second (F2) and 4 from the third (F3) from the strains A138, B147 and C23 were isolated. The D150 strain provied 29 F1 and F2 clones. The strains and clones had their isoenzyme and K-DNA patterns determined. The clones from A138, Bl47 and C231 strains presented isoemzyme and K-DNA patterns identical between thewmselves and their respective parental strains. Therefore showing the homogenety and stability of isoenzyme and K-DNA patterns after successive cloning. The Dl50 strain from zymodeme D (ZD) showed heterogeneity. Twenty-eight out of 29 clones of the first generation were of zymodeme A and only one was of zymodeme C, confirming previous reports that ZD strains consisted of ZA and ZC parasite populations. The only D150 strain clone of zymodeme C showed a K-DNA pattern identical to its parental strain. The remining clones although similar among themselves were different from the parental strain. Thus the T. cruzi strains had either homonogeneus or heterogeneous populations. The clones produced by successive cloning provided genetically homonogeous populations. Their experimental use will make future results more reliable and reproducible.

Kinetics of antigen specific and non-specific polyclonal B-cell responses during lethal Plasmodium yoelii malaria

In order to study the kinetics and composition of the polyclonal B-cell activation associated to malaria infection, antigen-specific and non-specific B-cell responses were evaluated in the spleens of mice infected with Plasmodium yoelii 17 XL or injected with lysed erythrocytes or plasma from P. yoelii infected mice or with P. falciparum culture supernatants. Spleen/body weigth ratio, numbers of nucleated spleen cells and Immunoglobulin-containing and Immunoglobulin-secreting cells increased progressively during the course of infection,in parallel to the parasitemia. A different pattern of kinetics was observed when anti-sheep red blood cell and anti-trinitrophenylated-sheep red blood cell plaque forming cells response were studied: maximum values were observed at early stages of infection, whereas the number of total Immunoglobulin-containing and Immunoglobulin-secreting cells were not yet altered. Conversely, at the end of infection, when these latter values reached their maximum, the anti-sheep red blood cell and anti-trinitrophenylated-sheep red blood cell specific responses were normal or even infranormal. In mice injected with Plasmodium-derived material, a higher increase in antigen-specific PFC was observed, as compared to the increase of Immunoglobulin-containing and Immunoglobulin-secreting cell numbers. This suggested a "preferential" (antigen-plus mitogen-induced) stimulation of antigen-specific cells rather than a generalized non-specific (mitogen-induced) triggering of B-lymphocytes. On the basis of these and previous results, it is suggested that polyclonal B-cell activation that takes place during the course of infection appears as a result of successive waves of antigen-specific B-cell activation.

Esquistossomose mansoni: tratamento medicamentoso

It is the specific treatment of mansoni schistosomiasis that aims to act directed on the parasite, through chemotherapy. Constitutes fundamental indication to the treatment of schistosomiasis active forms, that is, these determined by the presence of living eggs in the feces or in material from rectal biopsy, since eventual contra indications are respected. Two are the medicaments actually used: oxamniquine, used in the single dosage of 15mg/kg, V.O. for adults and 20mg/kg V.O. for children divided in two doses, offers a percentage of 30 to 40% of cures, evaluated by quantitative "oogram" and prazinquantel, in the single dose of 60 mg/kg V.O., presents a cure index of 30% however in seriate doses, of 60mg/kg during 3 days or 30mg/kg, 6 days, cure percentage is elevated to 95% evaluated by oogram. The evaluation of the treatment by quantitative or qualitative examination methods does not show the same sensibility. The percentage of cure according to feces examination, the quantitative of Kato-Katz or the qualitative (sedimentation), showed indexes from 90 to 100% for either one of the drugs, even in single dose, which evidences the difference of methodology of therapeutic evaluation. Tolorance to both medicaments is from good to regular, with collateral effects in 30 to 40% of the patients.

Trypanosoma cruzi: metacyclogenesis in vitro - I. Changes in the properties of metacyclic trypomastigotes maintained in the laboratory by different methods

In this work we have studied the modifications in the biological properties of Trypanosoma cruzi when the parasite is maintained for a long time in axenic culture. The studies were done with a clone from an avirulent strain (Dm30L) and a non-cloned virulent strain (EP) of T. cruzi. Both parasiteswere maintained, for at least three years, by successive triatomine/mouse alternate passage (control condition), or by serial passage in axenic medium (culture condition), or only in the mouse (mouse condition). The comparison between parasites of culture and control condition showed that metacyclogenesis capacity was reduced in the former and that the resulting metacyclics displayed an attenuatedvirulence. In order to compare the virulence of metacyclics from the urine of the insect vector, Rhodnius prolixus were infected by artificial feeding with parasites of the control or culture condition. After three triatomine/triatomine passages, there was observed an almost identical biological behavior for these parasites, hence indicating that the maintenance of T. cruzi for a long time in axenic culture affects the differentiation capacity and the virulence of the parasite. Additionally, it was demonstrated that it is possible to maintain T. cruzi exclusively through passages in the invertebrate host.

Effect of Niclosamide (Bayluscide WP 70 (R)), Anacardium occidentale hexane extract and Euphorbia splendens latex on behavior of Biomphalaria glabrata (Say, 1818), under laboratory conditions

The repellent effect of the molluscicides Niclosamide (Bayluscide WP 70 (R)), Anacardium occidentale and the latex of Euphorbia splendens on Biomphalaria glabrata was observed through the investigation of the occurrence of escape behavior among molluscs that were exposed to dosages lower than the LD 50. The total number of individuals out of water among the surviving snails in the control group provided a "Natural Escape Index". The comparison between this total and the total number of surviving snails in each group exposed to the different dosages of the molluscicides after 24 hr provided the "Molluscicide Escape Index" and the detection of a "Repellency Range" to these snails. The escape indexes for Niclosamide, A. occidentale and E. splendens were 10, 6.22 and 6.44 respectively. Repellency occurred at the following concentration ranges: 0.01, 0.02 and 0.03 ppm Bayluscide, 0.1, 0.2 and 0.3 ppm A. occidentale and 0.05, 0.10, 0.15 and 0.20 ppm E. splendens. The Natural Escape Index obtained in the control group was zero.

Ultrastructural aspects of spermatogonesis in Metamicrocotyla macracantha (Alexander, 1954) Koratha, 1955 (Monogenea; Microcotylidae)

Ultrastructural aspects of spermatogenesis, spermiogenesis and of the mature spermatozoon of a microcotylid monogenean Metamicrocotyla macracantha parasite from Mugil liza, are described. The irregularly-shaped spermatogonia divides by successive mitoses, forming the primary spermatocytes, identified by the presence of synaptonemal complexes in their nuclei. The spermatids formed by meiotic cell divisions of the secondary spermatocytes, differentiate into a mature spermatozoon. Cross sections of the head and the middle region of mature spermatozoa show the nucleus with strong condensed chromatin, the mitochondria with short cristae, peripheral microtubules and two axonemes with a 9+1 pattern, confirming the characteristics of this genus.

Characterization of subpopulations (clones and subclones) of the 21 SF strain of Trypanosoma cruzi after long lasting maintenance in the laboratory

Several studies have shown a clonal structure of Trypanosoma cruzi and its possible correlation with the behavioral heterogeneity of the parasite strains. In the present study, the 21 SF strain, that have been maintained in laboratory by successive passages in mice, for more than 15 years, showing a stability of biological and isoenzymic characteristics has been cloned, with the objective of establishing the characters of its clones and subclones. With the technique of isolation of a single parasite from the blood of infected mice, 5 clones and 14 subclones have been obtained. After four passages into mice, inoculum of 10(5) was obtained for each clone and subclone and inoculated into mice weighing 10 to 12 g. These were used for the study of the biological behavior of the clones: evolution of parasitemia, morphology of blood forms and host mortality. For isoenzymic characterization, the clones and subclones were analyzed for ALAT, ASAT, GPI and PGM enzymes. Results have shown that the 5 clones and the 14 subclones disclosed a biological behavior similar to the parental strain, with minor variability of the parasitemic profiles and also the same isoenzymic patterns. These results confirm the stability of the 21 SF strain and indicate a clonal homogeneity of its populations. This is compatible with the hypothesis that the T. cruzi strains represent an equilibrium of either homogenous or heterogeneous populations.

Trypanosoma cruzi: Maintenance in Culture Modify Gene and Antigenic Expression of Metacyclic Trypomastigotes

In this study we examined whether the maintenance of Trypanosoma cruzi by long-time in axenic culture produces changes in gene expression and antigenic profiles. The studies were made with a Dm30L-clone from a low-virulent strain and a non-cloned virulent EP-strain of T. cruzi. Both parasites were maintained, for at least seven years, by successive alternate passage triatomine/mouse (triatomine condition), or by serial passage in axenic medium (culture condition). The comparison of the [35S]methionine metabolic labeling products of virulent and non-virulent parasites by 2D-SDS-PAGE, clearly indicates that the expression of metacyclic trypomastigotes (but not of epimastigotes) proteins have been altered by laboratory maintenance conditions. Western blot analysis of EP and Dm30L-epimastigotes using a serum anti-epimastigotes revealed that although most of antigens are conserved, four antigens are characteristics of triatomine condition parasites and three other are characteristics of culture condition parasites. Anti-metacyclics serum revealed significative differences in EP- and Dm30L-metacyclic trypomastigotes from triatomine condition. However, avirulent metacyclic forms were antigenically very similar. These results suggest that besides a possible selection of avirulent subpopulation from T. cruzi strains genetically heterogeneous when maintained by long time in axenic culture, changes in virulence might be due to post-translational modifications of the antigens induced by the absence of the natural alternability (vertebrate-invertebrate) in the life-cycle of T. cruzi