Methylene blue vital staining for Trypanosoma cruzi trypomastigotes and epimastigotes

The morphological identification of Trypanosoma cruzi is currently considered to have a high specificity, but its sensitivity, which depends on the volume of the sample examined, is rather low. Trypanosome developmental stages suspended in blood, reduviid feces, and culture media are routinely searched for by means of fresh film examination (about 2 µL). High speed centrifugation of blood samples separates the buffy coat, where most trypomastigotes concentrate. As the parasites are transparent and colorless, their detection is mostly dependent on their motility. The fluorescent vital stain acridine orange has been used to enhance image contrast, as exemplified by the QBC (Quantitative Buffy Coat) technique. Staining blood, buffy coat, reduviid feces, and culture media samples with methylene blue (also a vital dye) is a means of producing sharp, well contrasted images of motile or non-motile T. cruzi developmental stages, only standard laboratory microscopes being required. Slides previously coated with a thin layer of methylene blue are used to stain fresh blood films. Photomicrographs exemplify the results of methylene blue staining applied to living and fixed parasites.

Genetic breeding on the bee Melipona scutellaris (Apidae, Meliponinae)

A selection of queens of Melipona scutellaris through the most productive colonies were carried out during eight months in an orange honeyflow. Each of the colonies was evaluated by its production, that is, the gross weight production ( pollen, brood, geopropolis and wax of each hive). With this data a coefficient of repeatability was estimated by the intraclass correlation method, obtained r = 0.835 ± 0.071. The repeatibility is very high showing that the analysed data (production) is repeatable. Selection was then carried out using the regression coefficient of each colony and the respective production gain. Using these data the colonies were divided into three groups according to the method Vencovsky and Kerr (1982): a with the colonies of highest productivity, b of least productivity, and c of intermediary productivity. Colonies with the highest production (Group a) gave their queens to those of the lowest production (Group b) after their queens were taken out and killed; while those of intermediate (Group c) stayed with the same queens during the entire experiment both before and after the selection. The modifications in weight, that is, the genetic response was (R)= 7.98 gr per day which indicated a selection gain. The estimate of the realized herdability is twice the rate of the response to selection (R) by the selection differential (S2). That is then h²R=2(R/S2) then h²R= 0.166

Determinação dos elementos-traço (Zn, Co e Ni) em sedimentos da Plataforma Continental Amazônica sob influência da descarga do rio Amazonas

Esta pesquisa foi desenvolvida na Plataforma Continental Amazônica (PCA) no trecho compreendido entre os cabos Orange (AP) e Maguari (PA), para determinar os teores de Zn, Co e Ni nos sedimentos superficiais, identificando o papel dos sedimentos como fonte ou reserva de elementos-traço. A concentração dos elementos metálicos apresentou a seguinte associação: Zn > Ni > Co. Os sedimentos localizados na PCA podem ser considerados como sítio de ocorrência natural dos elementos metálicos, não havendo influência de fontes poluidoras na liberação destes para o sistema.

Toxicity and repellency of essential oils to Zabrotes subfasciatus (Boheman) (Coleoptera, Chrysomelidae, Bruchinae) in Phaseolus vulgaris L

The effects of tangerine (Phaseolus vulgaris Blanco), lemon (Citrus medica limonum Lush), pear orange (Citrus sinensis L. Osbeck), red copaiba (Copaifera langsdorffii Desf.), rosemary (Baccharis dracunculifolia De Candole), Eucalyptus (Eucalyptus globulus Labillardière and E. citriodora Hook), lemongrass (Cymbopogon citratus Stapf.) and citronella (Cimbopogon nardus Linnaeus) oils at several concentrations on Zabrotes subfasciatus (Boheman) were studied. In toxicity tests, grains of Phaseolus vulgaris L. cv. Rajadinho were impregnated with oils and infested with adults of Z. subfasciatus up to 24 hours old. All tested oils were effective in reducing the viable egg-laying and adult emergence of this pest, in function of the concentrations used, highlighting E. citriodora and E. globulus oils which caused 100% effectiveness from 0.5 mL Kg-1 concentration. In repellency tests, two arenas consisting of plastic containers, connected symmetrically to a central box by two plastic tubes were used. In one of the boxes, untreated beans were placed and on the other ones beans treated with each oil concentration were used. In the central box, five couples of Z. subfasciatus were released. Grains of P. vulgaris treated with oils of E. citriodora, C. citratus and C. oleifera reduced the attraction percentage of Z. subfasciatus adults, while the E. globulus increased this percentage. The percentages of reduced viable eggs ranged from 17.9% (C. medica limonum) to 93.3% (C. nardus), while the reduction on the number of emerged insects was 23.9% and 95.9%, respectively for these same oils.

Estudos experimentais sôbre a origem do milho

1) It may seem rather strange that, in spite of the efforts of a considerable number of scientists, the problem of the origin of indian corn or maize still has remained an open question. There are no fossil remains or archaeological relics except those which are quite identical with types still existing. (Fig. 1). The main difficulty in finding the wild ancestor- which may still exist - results from the fact that it has been somewhat difficult to decide what it should be like and also where to look for it. 2) There is no need to discuss the literature since an excellent review has recently been published by MANGELSDORF and REEVES (1939). It may be sufficient to state that there are basically two hypotheses, that of ST. HILAIRE (1829) who considered Brazilian pod corn as the nearest relative of wild corn still existing, and that of ASCHERSON (1875) who considered Euchlaena from Central America as the wild ancestor of corn. Later hypotheses represent or variants of these two hypotheses or of other concepts, howewer generally with neither disproving their predecessors nor showing why the new hypotheses were better than the older ones. Since nearly all possible combinations of ideas have thus been put forward, it har- dly seems possible to find something theoretically new, while it is essential first to produce new facts. 3) The studies about the origin of maize received a new impulse from MANGELSDORF and REEVES'S experimental work on both Zea-Tripsacum and Zea-Euchlaena hybrids. Independently I started experiments in 1937 with the hope that new results might be obtained when using South American material. Having lost priority in some respects I decided to withold publication untill now, when I can put forward more concise ideas about the origin of maize, based on a new experimental reconstruction of the "wild type". 4) The two main aspects of MANGELSDORF and REEVES hypothesis are discussed. We agree with the authors that ST. HILAIRE's theory is probably correct in so far as the tunicata gene is a wild type relic gene, but cannot accept the reconstruction of wild corn as a homozygous pod corn with a hermaphroditic tassel. As shown experimentally (Fig. 2-3) these tassels have their central spike transformed into a terminal, many rowed ear with a flexible rachis, while possessing at the same time the lateral ear. Thus no explanation is given of the origin of the corn ear, which is the main feature of cultivated corn (BRIEGER, 1943). The second part of the hypothesis referring to the origin of Euchlaena from corn, inverting thus ASCHERSON's theory, cannot be accepted for several reasons, stated in some detail. The data at hand justify only the conclusion that both genera, Euchlaena and Zea, are related, and there is as little proof for considering the former as ancestor of the latter as there is for the new inverse theory. 5) The analysis of indigenous corn, which will be published in detail by BRIEGER and CUTLER, showed several very primitive characters, but no type was found which was in all characters sufficiently primitive. A genetical analysis of Paulista Pod Corn showed that it contains the same gene as other tunicates, in the IV chromosome, the segregation being complicated by a new gametophyte factor Ga3. The full results of this analysis shall be published elsewhere. (BRIEGER). Selection experiments with Paulista Pod Corn showed that no approximation to a wild ancestor may be obtained when limiting the studies to pure corn. Thus it seemed necessary to substitute "domesticated" by "wild type" modifiers, and the only means for achieving this substitution are hybridizations with Euchlaena. These hybrids have now been analysed init fourth generation, including backcrosses, and, again, the full data will be published elsewhere, by BRIEGER and ADDISON. In one present publication three forms obtained will be described only, which represent an approximation to wild type corn. 6) Before entering howewer into detail, some arguments against ST. HILAIRE's theory must be mentioned. The premendelian argument, referring to the instability of this character, is explained by the fact that all fertile pod corn plants are heterozygous for the dominant Tu factor. But the sterility of the homozygous TuTu, which phenotypically cannot be identified, is still unexplained. The most important argument against the acceptance of the Tunicata faetor as wild type relic gene was removed recently by CUTLER (not yet published) who showed that this type has been preserved for centuries by the Bolivian indians as a mystical "medicine". 7) The main botanical requirements for transforming the corn ear into a wild type structure are stated, and alternative solutions given. One series of these characters are found in Tripsacum and Euchlaena : 2 rows on opposite sides of the rachis, protection of the grains by scales, fragility of the rachis. There remains the other alternative : 4 rows, possibly forming double rows of female and male spikelets, protection of kernels by their glumes, separation of grains at their base from the cob which is thin and flexible. 8) Three successive stages in the reconstruction of wild corn, obtained experimentally, are discussed and illustrated, all characterized by the presence of the Tu gene. a) The structure of the Fl hybrids has already been described in 1943. The main features of the Tunicata hybrids (Fig. -8), when compared with non-tunicate hybrids (Fig. 5-6), consist in the absence of scaly protections, the fragility of the rachis and finally the differentiation of the double rows into one male and one female spikelet. As has been pointed out, these characters represent new phenotypic effects of the tunicate factor which do not appear in the presence of pure maize modifiers. b) The next step was observed among the first backcross to teosinte (Fig. 9). As shown in the photography, Fig. 9D, the features are essencially those of the Fl plants, except that the rachis is more teosinte like, with longer internodes, irregular four-row-arrangement and a complete fragility on the nodes. c) In the next generation a completely new type appeared (Fig. 10) which resembles neither corn nor teosinte, mainly in consequence of one character: the rachis is thin and flexible and not fragile, while the grains have an abscission layer at the base, The medium sized, pointed, brownish and hard granis are protected by their well developed corneous glumes. This last form may not yet be the nearest approach to a wild grass, and I shall try in further experiments to introduce other changes such as an increase of fertile flowers per spikelet, the reduction of difference between terminal and lateral inflorescences, etc.. But the nature of the atavistic reversion is alveadwy such that it alters considerably our expectation when looking for a still existing wild ancestor of corn. 9) The next step in our deductions must now consist in an reversion of our question. We must now explain how we may obtain domesticated corn, starting from a hypothetical wild plant, similar to type c. Of the several changes which must have been necessary to attract the attention of the Indians, the following two seem to me the most important: the disappearance of all abscission layers and the reduction of the glumes. This may have been brought about by an accumulation of mutations. But it seems much more probable to assume that some crossing with a tripsacoid grass or even with Tripsacum australe may have been responsible. In such a cross, the two types of abscission layer would be counterbalanced as shown by the Flhybrids of corn, Tripsacum and Euchlaena. Furthermore in later generations a.tu-allele of Tripsacum may become homozygous and substitute the wild tunicate factor of corn. The hypothesis of a hybrid origin of cultivated corn is not completely new, but has been discussed already by HARSHBERGER and COLLINS. Our hypothesis differs from that of MANGELSDORF and REEVES who assume that crosses with Tripsacum are responsible only for some features of Central and North American corn. 10) The following arguments give indirects evidence in support of our hypothesis: a) Several characters have been observed in indigenous corn from the central region of South America, which may be interpreted as "tripsacoid". b) Equally "zeoid" characters seem to be present in Tripsacum australe of central South-America. c) A system of unbalanced factors, combined by the in-tergeneric cross, may be responsible for the sterility of the wild type tunicata factor when homozygous, a result of the action of modifiers, brought in from Tripsacum together with the tuallele. d) The hybrid theory may explain satisfactorily the presence of so many lethals and semilethals, responsible for the phenomenon of inbreeding in cultivated corn. It must be emphasized that corn does not possess any efficient mechanism to prevent crossing and which could explain the accumulation of these mutants during the evolutionary process. Teosinte which'has about the same mechanism of sexual reproduction has not accumulated such genes, nor self-sterile plants in spite of their pronounced preference for crossing. 11) The second most important step in domestication must have consisted in transforming a four rowed ear into an ear with many rows. The fusion theory, recently revived byLANGHAM is rejected. What happened evidently, just as in succulent pXants (Cactus) or in cones os Gymnosperms, is that there has been a change in phyllotaxy and a symmetry of longitudinal rows superimposed on the original spiral arrangement. 12) The geographical distribution of indigenous corn in South America has been discussed. So far, we may distinguish three zones. The most primitive corn appears in the central lowlands of what I call the Central Triangle of South America: east of the Andies, south of the Amazone-Basin, Northwest of a line formed by the rivers São Prancisco-Paraná and including the Paraguay-Basin. The uniformity of the types found in this extremely large zone is astonishing (BRIEGER and CUTLER). To the west, there is the well known Andian region, characterized by a large number of extremely diverse types from small pop corn to large Cuszco, from soft starch to modified sweet corn, from large cylindrical ears to small round ears, etc.. The third region extends along the atlantic coast in the east, from the Caribean Sea to the Argentine, and is characterized by Cateto, an orange hard flint corn. The Andean types must have been obtained very early, and undoubtedly are the result of the intense Inca agriculture. The Cateto type may be obtained easily by crosses, for instance, of "São Paulo Pointed Pop" to some orange soft corn of the central region. The relation of these three South American zones to Central and North America are not discussed, and it seems essential first to study the intermediate region of Ecuador, Colombia and Venezuela. The geograprical distribution of chromosome knobs is rapidly discussed; but it seems that no conclusions can be drawn before a large number of Tripsacum species has been analysed.

A influência dos pigmentos amarelo-laranja da semente de milho na coloração da gema de ovo de galinha

The effect of carotenoid pigments on the egg yolk color was studied in this paper. Three types of maize of known genetical constitution were used: Cateto, with deep orange endosperm; Armour, with yellow-orange endosperm and Cristal, with white endosperm. The carotenoid pigments of the two colored maizes were analysed: the total and both the active parts in relation to vitamin A and the zeaxanthin part showed to be practically double in the deep orange corn. The color of the yolk was orange when the ration had the deep orange corn and yellow in the case of the yellow-orange corn. The increase in shade was proportional to the amount of pigment present in the grains. If green feeds is added to the ration with white corn, the yolk becomes yellow or orange, depending on the amount of green given to the chickens. The practical importance of controlling the color of the yolk was emphasized.

A influência dos pigmentos amarelo -laranja de vários alimentos na coloração da gema de ôvo de galinha

The effect of different feeds in comparison with that of maize grains on the egg yolk color was observed. It was found that deep orange and yellow orange maize give satisfactory coloration to the yolk, respectively orange and yellow. The most intense color was observed when green feed was used in combination with deep orange maize. Green feeds as chicory, alfafa, cabbage, welsh onion and banana leaves and alfafa or chicory meal proved to be good in giving orange color to the yolk. Yellow yolk was obtained when Guinea grass or carica fruit were used in the ration. Carrot and beet without leaves did not give satisfactory color to the egg yolk. The observations with other feeds are being continued.

Observações sôbre a quantidade de milho amarelo necessária para prevenir a avita-minose a em pintos

The authors tried to check in this experiment the minimum of yellow corn necessary for preventing avitaminosis A in chickens. It was observed, in balanced ration with 50% of corn, that: a) 20% of dent and yellow grains and 30% of flint and white grains were insuficient to prevent avitaminosis A. b) 20% of flint and orange grains and 30% of flint and white grains or 40% of either colored grains and 10% of flint, and white grains did not show evident signs of avitaminosis A during the 12 weeks of the experiment. The ration containing 20% of flint and orange grains is pratically equivalent to the ration containing 40% of dent and yellow grains, regarding the content of pro-vitamina A. However, it was not possible to conclude if these dosage are sufficient to give the necessary vitamina A for normal development of the chickens since the table 3 seems to indicate a negative correlation between the amount of pigment in the ration and the mortality of the animals.

Estudos sôbre a coloração da gema de ôvo de galinhas - IV. Efeito da raça e da quantidade de milho amarelo na ração

This paper deals with experiments on the yolk color of chicken eggs. The results obtained can be summarized as follows: a) no differences were observed when different breeds (S. C. W. Leghorn and Rhode Island Red) were fed whith the same ration: yellow corn grains or green feed. b) 30% of yellow corn grain (orange or yellow) in the ration are sufficient to give satisfactory color to the yolk eggs.

Influência de diferentes cavalos e da época de colheita no teor de vitamina C na laranja natal

This paper deals with the variation of the ascorbic acid content in Natal orange as related to the type of rootstock and to the picking time. The determinations were carried out in the juice of fruits which had practically the same ratio total solids/acidity being selected only the highest values of such quotient. The results can be summaryzed as follows: 1. The rootstock has a significant influence on the vitamin C content of the scion; this influence shows two interesting aspects: first, the ascorbic acid contet itself varies as a function of the rootstock; among the three rootstocks studied the highest values were obtained on scion growing on Serra d'água orange; the lowest values were found in "limão rugoso" and "limão francês" was intermediate; the second point to be emphasyzed is the manner of variation of the ascorbic acid content in the different rootstocks: in the case of Serra d'água. the content decreases slowly whereas in "limão francês" the level remains nearly constant and suddenly falls down; finally in the case of "limão rugoso" the amount of ascorbic acid increases significantly to return to the primitive level. 2. The amount of ascorbic acid, not considering the rootstock influence, varies strongly as a function of harvesting time; for this reason the present factor must be borne in mind when sampling for the determination of ascorbic acid in orange is concerned.

Oviposition preference hierarchy in Ceratitis capitata (Diptera, Tephritidae): influence of female age and experience

The influence of two factors, age and previous experience, on the oviposition hierarchy preference of Ceratitis capitata (Wiedemann, 1824) females was studied. Two populations were analyzed: one reared in laboratory during 17 years and the other captured in nature. In the first experiment the oviposition preference for four fruits, papaya, orange, banana and apple was tested at the beginning of oviposition period and 20 days past. The results showed that the wild females as much the laboratory ones had an oviposition preference hierarchy at the beginning of peak period of oviposition. However this hierarchic preference disappeared in a later phase of life. In the second experiment the females were previously exposed to fruits of different hierarchic positions and afterwards their choice was tested in respect to the oviposition preference for those fruits. The results showed that there was an influence of the previous experience on the posterior choice of fruits to oviposition when the females were exposed to fruits of lower hierarchic position.

Aceitação e preferência de frutos para oviposição em duas populações de Ceratitis capitata (Diptera, Tephritidae)

The influence of four host fruits, orange (Citrus sinensis L.), papaya (Carica papaya L.), mango (Mangifera indica L.) and apple (Malus domestica Borkh) on oviposition behavior of Ceratitis capitata (Wiedemann, 1824) was evaluated. Experiments were carried out on two C. capitata laboratory-reared populations: one with artificial diet for 10 years with periodic introduction of wild flies and other reared with artificial diet for 20 years without wild flies introduction. In acceptance experiments two pieces of a single fruit type were exposed to a group of 10 females; after 48 hours, these were changed by two new pieces of the same fruit type, and, in the end of the fourth day, the experiments were concluded. In preference experiments, two pieces of different fruits were offered to the females. These experiments were driven under the same conditions of the acceptance ones. Acceptance and preference experiments for both populations showed the following choice hierarchy: papaya > mango > orange > apple. The acceptance experiments showed no differences in the number of eggs/female/day laid in the first 48 hours and in the last 48 hours of the experiments. The amount of eggs in the two pieces of fruits offered to the females was similar. In spite of the two populations present similar behavior in relation to host hierarchy, the number of laid eggs was different, being larger for the population reared without wild flies introduction.

Descrição dos adultos de Camelobaetidius billi (Ephemeroptera, Baetidae)

The adults of Camelobaetidius billi Thomas & Dominique, 2000 are described for the first time. They can be distinguished from the other known adults of the genus by the following combination of characters: in males, turbinate portion of the compound eyes orange, abdominal color pattern, and the shape and relative length of the forceps articles; in females by the abdominal color pattern. The specimens were collected at Presidente Figueiredo, State of Amazonas, Brazil, and represent the first report of the species in Brazil.

A new species of Neosilba (Diptera, Lonchaeidae) from Brazil

A new species of Neosilba McAlpine, 1962, N. pradoi sp. nov., is described and illustrated. This new species was found in the south of Brazil (Rio Grande do Sul and Santa Catarina), in the southeast (State of São Paulo) and center west (State of Mato Grosso do Sul). It has been reared from fruits of guava (Psidium guajava, Myrtaceae), "araçá" (Psidium cattleyanum, Myrtaceae), "guabiroba" (Campomanesia xanthocarpa, Myrtaceae), Surinam cherry (Malpighia emarginata, Malpighiaceae), cherry (Prunus avium, Rosaceae), orange (Citrus sinensis, Rutaceae), "ingá" (Inga laurina, Fabaceae), "esporão-de-galo" (Celtis iguanae, Ulmaceae) and passion fruit (Passiflora edulis, Passifloraceae).

Sobre alguns casos de carcinoma primitivo do pulmão, do bronchio e da trachéa

1-Out of 1531 autopsies at Rio de Janeiro, Brazil, recorded in the Section of Pathology of «Instituto OSWALDO CRUZ», we found three cases of primary carcinoma of the lung, which gives the incidence of 0,195 per cent. They were not associated with tuberculos's and anthracos's was not marked. 2-The gross and microscopical features indicate that they are in reality primary cancers of the larger bronchi arising probably, from the bronchial lining epithelium. There is a striking similarity both macro and microscopical in Cases I and II, where the structure is that of cylindrical cell alveolar carcinoma. Mucus formation was never a prominent feature in these tumors. As in HENRICI's case they apparently invade the lung along the alveolar wall in more or less considerable extension retaining the alveolar septa as stroma. In case III the structure is that of squamous cell carcinoma, and the tumor mass, usually ulcerating, lies in the immediate vicinity of the bronchi, the extension to the lung tissue being not widespread. 3-Cases IV and V are tumors of the lower and median portion of the trachéa, squamous cell carcinoma in structure, which form marked contrast with the tumors originating in the larger bronchi: the tumor process never invades the lung parenchyma by continuous extension. In Case V extension to the mediastinal lymph-nodes is observed, and a tumor of the size of an orange is found in the anterior mediastinum: that tumor mass and the upper lobe of the right lung were only adherent by fibrous tissue of inflammatory origin and the tumor did not extend into the lung tissue.