Observaçães sôbre a congelação do plasma humano para desidratação

A technics for prefreezing of blood plasma and serum is described in this paper. The method indicated by Strumia et al. (2), uses a rapid local freezing to obtain the shell-freezing, with refigerated alcohol bath, at temperatures around minus 35ºC. On our work, it has been found that normal horse blood plasma fulfils the instructions given by Strumia, although normal human blood plasma, very often, fails to give the expected results. This is very disadvantageous at the routine work. With the use of small amounts of solid carbon dioxide, spread over the flasks, in the refrigerated bath, it has been possible to start the chrystallization. The technics prescribes a rapid cooling, like the one used by Strumia, to bring the temperature down, to about plus 10ºC. and, with rotating device stopped, the solid carbon dioxide is applied for one minute simultaneously on each flask. Starting rotation again, it begins to form a very uniform shell around the walls of the flasks.

A vegetação no município de Ilhéus Estado da Bahia: IV - Caracaterísticas analíticas e periodicidade sociológica

It is well-known that diverse groups of vegetation with an analogous but not identical floristic composition show an ecological similarity which leads to a determined type of vegetation. Starting from this fact it becomes evident that the scope of phytosociological work is the establishing of the significance of the species within the association and the discovery of the rules which govern associations. The floristic surveys made in the field have to be analysed statistically so as to obtain satisfactory results. The usefulness of this method depends largely on the possibilities of comparing the results with previous studies of the same kind, in the same country, or elsewhere. The method used in this paper is that of measuring circumferences and counting individuals in the different associations studied because it permits the presentation of the results in tables which show the phytosociological complexity of the Brazilian rain-forests. The classical method of characteristics is valuable because the more evolved an association is the less sociable are the species it contains, so that such groups do not show clear differences between species but rather between sinusia or strata of individuals. Five tables are presented in which several of the qualitative and quantitative characteristics are studied with a view to discussing their value in relation to groups and species. They are: I - Abundance (number of individuals). II - Density (distance in meters between the individuals belonging to each stratum). III - Basal coverture (circles measured in square meters of the mean projection of the individuals on the surface). IV - Basal area (mean circle in square meters occupied by individuals on the surface). V - Frequency, abundance and sociability in relation to occurrence. TABLE I. This table indicates first the number of individuals in relation to the sinusia studied: next, the sum total of the individuals belanging to the strata are given for each association, thus providing the numeric value of the sinusia: finally, the relation between the total number of individuals in the association and the total for the sinusium thus fursnishing the abundance per sinusium, in the classic sence, that is the percentage, or rather the relative number, of the elements which compose the group. CONCLUSIONS. The general character of abundance of the regional vegetation of Ilheos may be summed up in the following way: as an association evolves towards permanent equilibirum the number of individuals the inferior strata diminishes in relation to those of the superior strata which increase. For the shrub sinusium, two important facts were observed: a) in a given association the number of elements of the inferior strata diminishes as the diameter of the individuals increases; b) the percentage of individuals belonging to the shrub sinusium in the sere diminishes as the association evolve. In the subarboreal sinusium it is seen that: in the sere the number of subarboreal individuals does not vary much; whereas in the climax or the prisere there is a fall owing to the equilibrum of the biologic forms. In the arboreal sinusium the following conclusion can be deduced from analogous facts: the number of individuals of the lower strata diminishes as circumference increases. Also, in the sere there is a progressive sequence for the individuals belonging to the superior strata. TABLE II. The relation between the mean distances of individuals belonging to the same stratum and the area of an association is equal to the density. The table shows that the mean density of the association and the distance between the individuals belonging to the strata of the same sinusium in relation to the total number of individuals belonging to the association. CONCLUSIONS. As rule, the density of individuals in the associations studied follows a very general character or at least a regional one: the distance between the individuals in the diverse strata varies according to their abundance and sociability. Two other facts of some sociological importance are: a) in identical strata of the same sere the density of individuals oscillates in an analogous manner in teh subclimaces and varies in the stages which have reached equilibrium. b) the density of individuals varias in accordance with the sinusium and the distances between individuals of the same sinusium varg in accordance with the strata. TABLE III. This table presents the mean basal individual coverture, that is the mean projection of the frond of the various individuals belonging to the same stratum. The means were obtained by measurement in the field, of 100 individuals belonging to each stratum and their projection on the surface. In the latoratory these measurements were converted into mean circles (in square meters) and the result was multiplied by the individuals belonging to the strata corresponding to the sinusium of each association. The result obtained is named basal coverture. CONCLUSIONS. As a rule, the basal coverture of the vegetation of the county of Ilhéus indicates that: in the evolution of the vegetation the basal coverture of the arbustive sinusium diminishes progressively whereas that of the arboreal one increases. The special norms obtained are: 1) in the shrub sinusium the basal coverture seems to follow a uniform norm, that is, in stages of evolution of the subclimax the basal coverture oscillates with a certain uniformity. 2) in the subarboreal sinusium this fact is related to the vitality and age of the species, as in the subclimax the number of young trees is large and the vitality of the species very variable. This permits the conclusion that: in the sere the basal coverture increases with the evolution of the vegetation and diminishes when an equilibrium is reached. 3) in the tree sinusium the climax association of the prisere and subsere seem to obey a binomial rule, as the coverture (density-abundance) increases until a determined stratum is reached and...

Mitsuda's reactions: induced by BCG in the normal Rhesus ("Macacca mulatta")

The reversals of Mitsuda's reactions induced by BCG have been objected to based on the possiblem interference of other determination causes of the phenomenon: tuberculous primo-infections, communicants of unsuspected leprosy, revearsals due to other causes, such as anti-diphteric and anti-tetanic vaccination, etc. In order to study the problem, we have used Rhesus monkeys (Macaca mulatta), which were reared in isolation, in an attempt to avoid the referred to interferences. Prior to the experiments, all animals were tested and found negative to radiograph, tuberculin and lepromin tests and were then submitted to the application of BCG vaccine (from 1 to 3 days old), in different doses and by different via. At different times, after the application of BCG, they were again submitted to the radiographic, tuberculin and lepromin tests. In the tables I to IV the experiences were summarised. From the experiments, the following conclusions were reached: 1 - From 12 Rhesus that received BCG 11 showed reversals of the Mitsuda reaction (91.7%). 2 - These reverseals took place both in tests effected shortly after BCG (from 6 days to 2 months), and tests effected much later (from 7 to 12 months after BCG). 3 - Some differences were found in the results, according to the dosis and the application via of the BCG. a) - The testicular and peritonela via (0,02g) were the only that determined strong positive Mitsuda's reactions (+++). b) - By oral via, animals that received high dosis (0.6g and 1.2 g), there resulted uniform and regular reversals, even though of low intensity (+); but from those who got small doses (0.2 g.) one showed no reversals in all tests, and the other presented reversals in the 2nd and 3rd tests only, also with low positivity (+). 4) In the 2nd and 3rd Mitsuda's reactions in the same animals, positivity was always precocious (generally within 48 hours), one getting the impression that there occurs a sensibilization of the animal body by the antigen with the repetition of the tests, even though the intensity of the reaction always remains the same. This precocious reaction (Fernandez type) occurs both shortly and long time after the application of the BCG. Its precocity depends not of the antigen only because the first Mitsuda's reaction after the BCG application occurs after some time and seems not influenced by the control lepromin test effected on the Rhesus before the BCG. 5) On the control group, the animals which received a.a.f. bacilli suspensions (Mycobacterium sp.; M. avium, and M. smegmatis), did not show reverseals of the Mitsuda's reaction. Two Rhesus, however, which received dead BCG (120ºC autoclave 1 hour), one intradermically (0.006 g) and the other orally (1.2 g), did both present reversals of the Mitsuda's reaction, with weak positivity (+). In all animals of the control-group, the allergic reactions were found negative. 6) Strong local inflammatory reactions were observed in the Rhesus that had received living BCG by intradermal via, and in the one submitted to multipunctures, there occurred the formation of a large caseous abcess. 7) The allergic tuberculinic and infratuberculinic reactions appeared dissociated from the Mitsuda's reactions: sometimes they are more precocious, occurring before of the lepromin test; on other occasions they disappear, when the Mitsuda's reactions still persist; and finally, they may be absent, when the latter occur, especially after the oral application of the BCG. 8) In Rhesus which received BCG by testicular and peritonela via, in the infratuberculinic test (0.1 ml of total BCG extract), besides the classic answer, which occurs between 48 and 96 hours, one could observe a delayed answer (15 to 20 days), represented by a non-erythematous nodule, which persists for 11-14 days.

Miocardite no macaco Cebus após inoculações repetidas com Schizotrypanum cruzi

Transmission of Chagas disease is realized through contamination of ocular conjunctiva, mucosa or skin with infected dejections eliminated by the insect vectors of Schizotrypanum cruzi (Triatoma infestans, Panstrongylus megistus and Rhodnius prolixus). The triatomid bugs live in holes and craks in the walls, in beds, behind trunks, etc. Found in primitive mud huts covered with thatched roofs, and so the human dwellers have many chances to contract the disease, reinfections being reasonably more to expect than a single inoculation. Experimental work reproducing those natural conditions is welcomed as some important features in the pathologic picture of the disease such as the extensive myocardial fibrosis seen in chronic cases are still incompletely known. Microscopic changes were studied in the heart muscle of seven Cebus monkeys infected by S. cruzi. This animal survives the acute stage of the disease and so is particularly suited to experiments of long duration in which several inoculations of S. cruzi are performed. Three different strains of S. cruzi isolated from acute cases of Chagas' disease were employed. One monkey was injected in the skin with infected blood and necropsied after 252 days. Two monkeys were three times, and one, eight times infected in skin, one of them with contaminated blood, and two with contaminated blood and dejections from infected bugs. The necropsies were performed after 35, 95 and 149 days. One monkey was three times inoculated through the intact ocular conjunctiva (one time with infected blood, two times with dejections from infected bugs), and one time through the wounded buccal mucosa, and necropsied after 134 days. Another monkey was six times inoculated, four times through the intact ocular conjunctiva (one time with contaminated blood, three times with dejections from infected bugs) and two times injected in the skin with infected blood, and necropsied after 157 days. Finally, another monkey was nine times inoculated, four times through the intact ocular conjunctiva (one time with infected blood, and three times with dejections from infected bugs), and five times injected in the skin (four times with contaminated blood, and one time with dejections from infected bugs), and necropsied after 233 days. The microscopic picture was uniform presenting, however, considerable individual variations, and was represented by diffuse interstitial myocarditis, frequently more (marked in the right ventricle base of the heart), accompanied by lymphatic stasis. The infiltration consists of macrophages, plasma cells and lymphocytes, the cellular reaction having sometimes a perivascular distribution, involving the auriculo-ventricular system of conduction, endocardium, epicardium and cardiac sympathetic gangliae. The loss of cardiac muscle fibers was always minimal. Leishmanial forms of S. cruzi in myocardial fibers are scanty and, in two cases, absent. Fatty necrosis in the epicardium was noted in two cases. Obliterative changes of medium-sized branches of coronary arteries (hypersensitivity reaction?) and multiple infarcts of the myocardium was found in one instance. The diffuse myocarditis induced by S. cruzi in several species of monkeys of the genus Cebus observed after 233 days (several inoculations) and 252 days (single inoculation) is not associated with disseminated fibrosis such as is reported in chronic cases of Chagas' disease. Definite capacity of reversion is another characteristic of the interstitial myocarditis observed in the series of Cebus monkeys here studied. The impression was gained that repeated inoculation with S. cruzi may influence the myocardial changes differently according to the period between the reinoculations. A short period after the first inoculation is followed by more marked changes, while long periods are accompanied by slight changes, which suggests an active immunisation produced by the first inoculation. More data are required, however before a definite statement is made on this subject considering that individual variations, the natural capacity of reversion of the interstitial myocarditis and the employement of more than a species of Cebus monkeys probably exerts influence also in the results here reported.

Os grandes clímaces do Brasil: II - considerações gerais sôbre a vegetação da região amazônica

Um eine neue phytogeographische Einteilung des Amazonasgebietes (Região Amazônica) in Angriff nehmen zu koennen, zieht der Autor anfangs die Ergebnisse der geomorphologischen, klimatologischen und floristischen Studien aelterer und neuerer Beobachter heran und setzt sie in Uebereinstimmung mit den oekologischen Resultaten. Die Einteilung umfasst: A. Region des guianensischen Schildes mit 5 Subregionen; B. Region der Sedimentboeden mit 3 teilweise unterteilten Subregionen; C. Region des nordbrasilianischen Schildes mit 2 unterteilten Subregionen. In den drei Gebieten finden wir, infolge der verschiedenen geologischen Verhaeltnisse, praktisch alle Vegetationstypen Brasiliens, obwohl das Klima uniform (warm-feucht) ist. Nach der Beurteilung der verschiedenen Theorien ueber die geologischen und floristischen Gegebenheiten und unter Hinzufuegung biologischer Tatsachen (Samenverbreitung) kommt der Autor zum Schluss, dass in dem Gebiet vier grosse, endemisch begruendete Baumzonen bestehen, deren Existenz auf dem heutigen hydrographischen Netz und seinen oekologischen Bedingugen beruht: 1. Die Trockengebiete der kristallinen Hochebenen am oberen Rio Negro und Rio Branco und auf dem suedamazonischen Schild; 2. Eine die Sedimentregion umgebende Zone; 3. Die Gebiete der Hochebenen und die hoeher gelegenen Teile der Sandaufschuettungen; 4. Die fruchtbaren Sedimentebenen. Ausserdem kann man die wechselnden Gebiete periodischer Ueberschwemmungen als kennzeichnende Zonen der augenblicklichen Entwicklungsphase des Amazonasbeckens ansehen.

Catálogo sistemático dos pólens das plantas arbóreas do brasil meridional: V - Leguminosae: Papilionatae

In this paper of the catalogue of south brazilian arboreal pollen grains, the autor deals with the Papilionatae. The Mimosoideae and Caesalpinioideae are yet in preparation, so that a discussion of the three subfamilies (or families) is not possible. In relation with the systematical subdivision of the Papilionatae, we found a large correspondence with the morphology of the present pollen grains. The group of Phaseoleae contains the genera Mucuna, Erythrina and Dioclea; the grains of the studied species are very different one from another; the first of the genera possesses very volumous grains, with three colpori and a reticulated superficies; the second has three-porated pollen grains with a large reticulated superficies, and the third, Dioclea, is yet different; it possesses oblated grains, each three-colporated, with a thick sexine and a psilated superficies. So, we can say, that Phaseoleae is a erypalynous group. Dalbergieae, with the genera: Andira, Dalbergia, Lonchocarpus, Machaerium, Platymiscium and Pterocarpus (and Dahlstedtia, the only exception), has very uniform pollen grains, and may be considered stenopalynous. It is not possible to include the genus Dahlstedtia into this group. A little exception is represented by Pterocarpus violaceus, because of the reticulated sexine of its grains, while the others, also three-colporated, possess a tectate-reticulated sexine. The genera Myrocarpus and Ormosia, from Sophoreae, are very more similar to the Dalbergieae as to any other genus of the Phaseoleae.

Factors influencing the development of Plasmodium gallinaceum in Aedes fluviatilis

Aedes fluviatilis is susceptible to infection by Plasmodium gallinaceum and is a convenient insect host for the malaria parasite in countries where Aedees aegypti cannot be maintained in laboratories. In South America, for instance, the rearing of A. aegypti the main vector of urban yellow fever, is not advaisable because of the potential health hazard it represents. Our results of the comparative studies carried out between the sporogonic cycle produced with two lines of P. gallinaceum parasites into A. fuviatilis were as follows. As proved for A. aegypti, mosquito infection rates were variable when A. fluviatilis blood-fed on chicks infected with and old syringe-passaged strain of P. gallinaceum. Oocysts developed in 41% of those mosquitos and the mean peak of oocyst production was 56 per stomach. Salivary gland infections developed in about 6% of the mosquitos. The course of sporogony was unrelated to the size of the inoculum administered to chicks or to the route by which the birds were infected. The development of infected salivary glands was unrelated to oocyst production. Sporogony of P. gallinaceum was more uniform when mosquitos blood-fed on chicks infected with a sporozoite-passaged strain. Oocysts developed in about 50% of those mosquitoes and the mean peak of oocyst production was 138 per stomach, with some individuals having as many as 600-800 oocysts. Infected salivary glands developed in a mean of 27% of the mosquitos but, in some batches, was a high as 50%. Patterns of salivary gland parasitism were similar to those of oocyst production. The course of sporogony of P. gallinaceum in A. fluviatilis is analized in relation to degree of parasitemia and gametocytemia in the vertebrate host.

Competitive interactions between species of freshwater snails: I. Laboratory: Ia. General methodology

For the development of laboratory experiments on the competitive interacitons between freshwater snail populations, special snail rooms were set up in the main building of the Research Center "Aggeu Magalhães". In the current paper, the first of a series on this subject, the general methodology of the laboratory work is described in detail. Using indoor cement channels in which a uniform seminatural environment was created, interactions of freshwater snail populations can be studied with minimal interference of the usual variables. Controlled indoor environmental techniques, as described in the current paper, may also be utilized in different types of experiments in malacology, and represent a substantial technical advance in malacological work.

Digamacris n. gen. (Orthoptera, Acrididae, Melanoplinae) de la region atlantica meridional de Brasil

Digamacris, a new genus of the Dichroplini (Acrididae, Melanoplinae) is described for the species Pezotettix amoenus Stal, 1878 and Dichroplus fratemus Carl, 1916, both included at present in the genus Dichroplus. These species live in edges and clearings of the Atlantic Forest (Mata Atlantica) of Brazil. D. fratemus is found in the states of Minas Gerais, Espirito Santo and Rio de Janeiro east of the Bay of Guanabara. D. amoenus in the state of Rio de Janeiro W. of the Bay of Guanabara and in the coastal area of the state of São Paulo. The species are illustrated and redescribed. Both have two neatly different chromatic forms of females, while the males are uniform in coloration and closely correspond with one of the female forms.

An improved silver staining procedure for schizodeme analysis in polyacrylamide gradient gels

A simple protocol is described for the silver staining of polyacrylamide gradient gels used for the separation of restriction fragments of kinetoplast DNA [schizodeme analysis of trypanosomatids (Morel et al., 1980)]. The method overcomes the problems of non-uniform staining and strong background color which are frequently encountered when conventional protocols for silver staining of linear gels. The method described has proven to be of general applicability for DNA, RNA and protein separations in gradient gels.

Gastromermis cordobensis n. sp. (Nematoda: Mermithidae) parasitizing Simulium lahillei Patterson & Shannon (Diptera: Simuliidae) in Argentina

Gastromermis cordobensis n. sp (Nematoda: Mermithidae) a parasite of larvae of the blackfly Simulium lahillei Paterson & Shannon (Diptera: Simuliidae) in Argentina, is described. Diagnostic characters of this species include a mouth ventrlly shifted; six cephalic papillae; eigh hypodermal chords; small and pear shaped amphids; a long and S-shaped vagina; a singl spicule, which is long, has non-uniform walls, and a tip with sculpture; three rows of genital papillae, the middle one with 18 pre-anal and 10 post-anal papillae, the lateral rows have 36 papillae each; oval eggs; and post-parasitic juveniles with long thin tails. Pre-parasitic and parasitic juveniles are included in the description.

Molecular approaches to malaria and Babesisosis diagnosis

The development of additional methods for detecting and identifuing Babesia and Plasmodium infections may be useful in disease monitoring, management and control efforts. To preliminarily evaluate sunthetic peptide-based serodiagnosis, a hydrophilic sequence (DDESEFDKEK)was selected from published BabR gene of B. bovis. Immunization of rabbits and cattle with the hemocyanin-conjugated peptide elicited antibody responses that specifically detected both P. falciparum and B. bovis antigens by immunofluorescence and Western blots. Using a dot-ELISA with this peptide, antisera from immunized and naturally-infected cattle, and immunized rodents, were specifically detected. Reactivity was weak and correlated with peptide immunization or infection. DNA-based detection using repetitive DNA was species-specific in dot-blot formats for B. bovis DNA, and in both dot-blot and in situ formats for P. falciparum; a streamlined enzymelinked synthetic DNA assay for P. falciparum detected 30 parasites/mm(cúbicos) from patient blood using either colorimetric (2-15 h color development) or chemiluminescent detection (0.5-6-min. exposures). Serodiagnostic and DNA hybridization methods may be complementary in the respective detection of both chronic and acute infections. However, recent improvements in the polymerase chain reaction (PCR) make feasible a more sensitive and uniform approach to the diagnosis of these and other infectious disease complexes, with appropriate primers and processing methods. An analysis of ribosomal DNA genes of Plasmodium and Toxoplasma identified Apicomplexa-conserved sequence regions. Specific and distinctive PCR profiles were obtained for primers spanning the internal transcribed spacer locus for each of several Plasmodium and Babesia species.

Structure and diversity of endoparasitic infracommunities and the trophic level of Pseudoplatystoma corruscans and Schizodon borelli (Osteichthyes) of the high Paraná river

One hundred and ten specimens of Pseudoplatystoma corruscans (Pimelodidae) and 582 specimens of Schizodon borelli (Anostomidae) collected in the high Paraná River were analyzed. On necropsy 74% of P. corruscans were found to be parasitized; proteocephalidean cestodes presented the greatest number. With regard to S. borelli, the percentage of parasitism reached 19.42% and the nematode Cucullanus pinnai was the most abundant. The absence of correlation between the endoparasitic diversity and the standard length of the two host species indicates that each one presents homogeneity in alimentary behaviour during all its life time, permiting the uniform recruitment of the same species of endoparasites during all its ontogenetic development. Independence of diversity values in relation to sex of P. corruscans and S. borelli evidences that the ecological relationships are similar between males and females in these species. Both host's infrapopulations presented a typical overdispersed pattern of distribution with isolationist characteristics.

The Trypanosoma cruzi Genome Project: Nuclear Karyotype and Gene Mapping of Clone CL Brener

By using improved pulsed field gel electrophoresis conditions, the molecular karyotype of the reference clone CL Brener selected for Trypanosoma cruzi genome project was established. A total of 20 uniform chromosomal bands ranging in size from 0.45 to 3.5 Megabase pairs (Mbp) were resolved in a single run. The weighted sum of the chromosomal bands was approximately 87 Mbp. Chromoblots were hybridized with 39 different homologous probes, 13 of which identified single chromosomes. Several markers showed linkage and four different linkage groups were identified, each comprising two markers. Densitometric analysis suggests that most of the chromosomal bands contain two or more chromosomes representing either homologous chromosomes and/or heterologous chromosomes with similar sizes

Internalization of components of the host cell plasma membrane during infection by Trypanosoma cruzi

Epimastigote and trypomastigote forms of Trypanosoma cruzi attach to the macrophage surface and are internalized with the formation of a membrane bounded vacuole, known as the parasitophorous vacuole (PV). In order to determine if components of the host cell membrane are internalized during formation of the PV we labeled the macrophage surface with fluorescent probes for proteins, lipids and sialic acid residues and then allowed the labeled cells to interact with the parasites. The interaction process was interrupted after 1 hr at 37ºC and the distribution of the probes analyzed by confocal laser scanning microscopy. During attachment of the parasites to the macrophage surface an intense labeling of the attachment regions was observed. Subsequently labeling of the membrane lining the parasitophorous vacuole containing epimastigote and trypomastigote forms was seen. Labeling was not uniform, with regions of intense and light or no labeling. The results obtained show that host cell membrane lipids, proteins and sialoglycoconjugates contribute to the formation of the membrane lining the PV containing epimastigote and trypomastigote T. cruzi forms. Lysosomes of the host cell may participate in the process of PV membrane formation.