Aspectos bioquímicos y celulares de la reproducción de los vectores de la enfermedad de Chagas: regulación e importancia fisiológica. Biochemical and cellular aspects of the reproduction of Chagas’ disease vectors: regulation and physiologycal impact

En la hipótesis de trabajo del presente proyecto se considera la importancia del metabolismo de lípidos y proteínas en los insectos hematófagos, en particular en los vectores de la enfermedad de Chagas, para afrontar exitosamente la demanda energética de la reproducción. Las hembras de estas especies pueden ingerir una comida de sangre abundante en lípidos y proteínas, los que son modificados en el intestino para su utilización y posterior almacenamiento en estructuras organizadas en el tejido ovárico, sustentando así el rápido crecimiento de los ovocitos. Estos aspectos resultan críticos para el ciclo de vida del insecto y para el mantenimiento de la cadena epidemiológica de la enfermedad. En estas especies, recientemente hemos caracterizado a nivel bioquímico y celular la interacción entre lipoproteínas y tejidos [Fruttero y col., Insect Biochem. Mol. Biol. 39: 322-331 (2009); Fruttero y col. Biocel 33 (3): 260 (2009)] y las fases del ciclo reproductivo [Aguirre y col., J. Insect Physiol. 54: 393-402 (2008)]. No obstante, los factores que participan en su regulación son aún escasamente conocidos. En este contexto, el estudio propone emplear dos especies de triatominos con el objeto de: (1) caracterizar los factores involucrados en la formación y regulación de reservas nutricionales en los ovocitos; (2) analizar los eventos que participan en la regresión del tejido ovárico: atresia folicular y mecanismos de muerte celular. (3) evaluar el impacto de productos naturales (ureasas vegetales y péptidos derivados) en el desarrollo del tejido ovárico. Para la ejecución de los objetivos se llevarán a cabo ensayos in vivo e in vitro con trazadores fluorescentes, fraccionamiento subcelular, estudios de expresión de proteínas (mRNA y proteína), estudios histo-morfológicos, ultraestructurales e inmunocitoquímicos, microscopía láser confocalizada, ensayos de actividad enzimática, ELISA, western-blot, electroforesis bidimensional, espectrometria de masas en tándem, etc. También se evaluarán los mecanismos de muerte celular (apoptosis/autofagia) mediante microscopía electrónica, detección de apoptosis in situ (TUNEL), inmunofluorescencia, etc. Los resultados obtenidos permitirán un mejor conocimiento sobre la fisiología y bioquímica de estos vectores, los que resultan indispensables en el diseño de nuevas estrategias para su control. Debido a la carencia de un tratamiento específico para la enfermedad y a la falta de métodos preventivos (vacuna), el control del vector es una de las vías más importantes para reducir la incidencia de la enfermedad. Actualmente, la situación socio-económica que sufren amplios núcleos de nuestra población propicia condiciones de vida que facilitan la reproducción de los vectores y la transmisión vectorial del parásito. El estudio permitirá además explorar aspectos bioquímicos y celulares básicos, generando conocimientos que podrían ser extensivos a otros insectos de importancia económica en la ganadería y/o agricultura. The aim of this project is to analyze the biochemical and cellular events involved in the lipid and protein metabolism in Chagas' disease vectors, and to evaluate their impact on the physiology of reproduction, particularly in the formation of nutritional resources in developing oocytes. At present, little is known about these critical aspects for the life cycle of the insect and for the epidemiology of the disease. The experimental approaches, which will be carried out using two species of triatomines, were designed: (1) to characterize factors involved in the formation and regulation of nutritional resources in developing oocytes; (2) to analyze the biochemical and cellular events that play a role during the regression of ovarian tissue, including the processes of oocyte resorption and programmed cell death. (3) to evaluate the impact of natural products (ureases from jackbean and related peptides) in the development of ovarian tissue. Methods and techniques involved in the project are: in vivo and in vitro assays with fluorescent tracers, ELISA, chemical assays, enzyme activities, western-blot; protein expression (mRNA), histological techniques, immunohistochemical and ultrastructural studies. Cell death will be analyzed by detection of apoptosis in situ (TUNEL), immunofluorescence (for autophagy), among others. The results obtained from the study will offer the opportunity to explore important aspects in the biology and physiology of Chagas' disease vectors that could be of potential utility in designing alternative strategies for the control of the insect.

Automating the testing process of image processing algorithms

As digital imaging processing techniques become increasingly used in a broad range of consumer applications, the critical need to evaluate algorithm performance has become recognised by developers as an area of vital importance. With digital image processing algorithms now playing a greater role in security and protection applications, it is of crucial importance that we are able to empirically study their performance. Apart from the field of biometrics little emphasis has been put on algorithm performance evaluation until now and where evaluation has taken place, it has been carried out in a somewhat cumbersome and unsystematic fashion, without any standardised approach. This paper presents a comprehensive testing methodology and framework aimed towards automating the evaluation of image processing algorithms. Ultimately, the test framework aims to shorten the algorithm development life cycle by helping to identify algorithm performance problems quickly and more efficiently.

Methode zur integrierten ökologischen und ökonomischen Bewertung von Produktionsprozessen und -technologien

Assessment, environmental management, production planning, life cycle assessment, costs, activity based costing, inetragtion, model, process, technology

Vergleich der Umweltverträglichkeit neuer Technologien im Strassenverkehr

Life-cycle assessment, cost-benefit analysis, alternative powertrain systems, alternative fuels, traffic, transport, exhaust, CO2, climate protection

Estudos sôbre a alimentação mineral da mandioca (Manihot utilissima Pohl.)

1. The present work was carried out to study the effects of mineral nutrients in the yield as well as in the composition of cassava roots. The variety "Branca de Sta. Catarina" was grown by the sand culture method, the following treatments being used: N0 P0 K0, N0 P1 Kl, N1 P0 K1, N2 P1 K0, N2 P1 K1, N1 P2 K1, and N1 P1 K2, where the figures 0, 1, and 2 denote the relative proportion of a given element. The nutrients were given as follows: N = 35 grams of ammonium nitrate per pot loaded with 120 pounds of washed sand; P1 = 35 grams of monocalcium phosphate; Kl = 28 grams of sulfate of potash. Besides those fertilizers, each pot received 26 grams of magnesium sulfate and weekly doses of micronutrients as indicated by HOAGLAND and ARNON (1939). To apply the macronutrients the total doses were divided in three parts evenly distributed during the life cycle of cassava. 2. As far yield of roots and foliage are concerned, there are a few points to be considered: 2.1. the most striking effect on yield was verified when P was omitted from the fertilization; this treatment gave the poorest yields of the whole experiment; the need of that element for the phosphorylation of the starchy reserves explains such result; 2.2. phosphorus and nitrogen, under the experimental conditions, showed to be the most important nutrients for cassava; the effect of potassium in the weight of the roots produced was much less marked; it is noteworthy to mention, that in absence of potassium, the roots yield decreased whereas the foliage increased; as potassium is essential for the translocation of carbohydrates it is reasonable to admit that sugars produced in the leaves instead of going down and accumulate as starch in the roots were consumed in the production of more green matter. 3. Chemical analyses of roots revealed the following interesting points: 3.1. the lack of phosphorus brought about the most drastic reduction in the starch content of the roots; while the treatment N1 P1 K1 gave 32 per cent of starch, with NI PO Kl the amount found was 25 per cent; this result can be explained by the requirement of P for the enzymatic synthesis of starch; it has to be mentioned that the decrease in the starch content was associated with the remarkable drop in yield observed when P was omitted from the nutrient medium; 3.2. the double dosis of nitrogen in the treatment N2 P1 K1, gave the highest yields; however the increase in yield did not produce any industrial gain: whereas the treatment N1 P1 K1 gave 32 per cent of starch, by raising the N level to N2, the starch content fell to 24 per cent; now, considering the total amount of starch present in the roots, one can see, that the increase in roots yield did not compensate for the marked decrease in the starch content; that is, the amount of starch obtained with N1 P1 K1 does not differ statistically from the quantity obtained with N2 P1 K1; as far we know facts similar to this had been observed in sugar beets and sugar cane, as a result of the interaction between nitrogen and sugar produced; the biochemical aspect of the problem is very interesting: by raising the amount of assimilable nitrogen, instead of the carbohydrates polymerize to starch, they do combine to the amino groups to give proteinaceous materials; actually, it did happen that the protein content increased from 2.91 to 5.14 per cent.

Estudos sôbre alimentação mineral do cafeeiro: VIII. Estudo da absorção e da translocação do radiozinco no cafeeiro (Coffea arabica L.)

WATER-CULTURE EXPERIMENTS. Two water-culture experiments were carried out to study the absorption and the translocation of radiozinc in young coffee plants as influenced by two factors, namely, concentration of heavy metals (iron, man ganese, copper and molybdenum) and method of application. Inert zinc was supplied at an uniform rate of 0. 05 p. p. m.; the levels of iron supply were 0, 1.0, and 10.0 p. p.m.; manganese was supplied in three doses 0, 0.5, and 5.0 p. p.m.; copper- 0, 0. 02, and 0. 2 p. p. m.; molybdenum- 0, 0. 01, and 0. 1 p. p. m. When applied to the nutrient solution the activity os the radiozinc (as zinc chloride) was 0. 15 microcuries per plant. In the study of the leaf absorption, Zn65 was supplied at the level of 0. 10 microcuries per plant; in this case the radioative material was brushed either on the lower or on the upper surface or both two pairs of mature leaves. The absorption period was 8 weeks. The radioactivity assay showed the following results: 1 - Among the heavy metals herein investigated the iron concentration did not affect the uptake of the radiozinc; by raising the level of Mn, Cu and Mo ten times, the absorption dropped to 50 per cent and even more when compared with the control plants; when, however, these micronutrients were omitted from the nutrient solution, an increase in the uptake of zinc was registered in the minus Cu treatment only. The effects of high levels of Mn, Cu and Mo probably indicate an interionic competition for a same site on a common binding substance in the cell surface. 2 - The absorption of the radiozinc directly applied to the leaf surface reached levels as high as 8 times that registered when the root uptake took place. Among the three methods of application which have been tried, brushing the lower surface of the leaves proved to be the most effective; this result is easily understood since the stomatal openings of the coffee leaves an preferentially located in the lower surface - in this treatment, about 40 per cent of the activity was absorved and around 12 per cent were translocated either to the old or to the newer organs. Chemical analyses for heavy metals, were carried out only in the plants received Zn65Cl2 in the nutrient solution; the results were as follows; 1 - Control plants had, per 1,000 gm, of dry weight the following amounts in mg.: Zn- 48 in the roots and 29 in the tops; Fe- 165 in the roots and 9 in the tops; Mn- 58 in the roots and 15 in the tops, Cu- 15 in the roots and 1. 2 in the tops; Mo- 2. 8 in the roots and 0. 45 in the tops. 2 - The effect of different levels of micronutrients in the composition of the plants can be summarized as follows: Fe and Zn- when omitted from the nutrient solution, the iron and zinc contents in the roots decreased, no variation being noted in the tops; the higher dosis caused an accumulation in the roots but no apparent effect in the tops; Mn- by omitting this micronutrient a decrease in its content in the roots was noted, where as the concentration in the tops was the same; Mo- no variation in roots and tops contents when molybdenum was omitted; higher dosis of manganese and molybdenum increased the amounts formed both in the roots and in the tops. 3 - The influence of the different concentrations of micronutrients heavy metals on the zinc content of the coffee plants can be described by saying that: Fe and Mo- no marked variation; Mn- no effect when omitted, reduced amount when the high dosis was supplied; Mn- when the plants did not receive manganese the zinc content in roots and tops was the same as in the control plants; a decrease in the zinc content of the total plant occurred when the high dosis was employed; Cu -the situation is similar to that described for manganese. Hence, results showed by the chemical analyses roughly correspond to those of the radioactivity assay; the use of the tracer technique, however, gave best informations along this line. SOIL-POTS EXPERIMENTS. The two types of soils which when selected support the most extensive coffee plantations in the State of São Paulo, Brazil: "arenito de Bauru", a light sandy soil and "terra roxa legitima", a red soil derived from basalt. Besides NPK containing salts, the coffee plants were given two doses of inert zinc (65 and 130 mg ZnCl2 per pot) and radiozinc at a total activity of 10(6) counts/minute. The results of the countings can be summarized as follows: 1 - When plants were grown in "arenito de Bauru" the activity absorbed as per cent of the total activity supplied was not affected by the dosis of inert zinc. The highest value found was around 0. 1 per cent. 2 - For the "terra roxa" plants, the situation is almost the same; there was, however, a slight increase in the absorption of the radiozinc when 130 mgm of ZnClg2 was given: a little above 0. 2 per cent of the activity supplied was absorbed. The results clearly show that the young coffee plants practically did not absorb none of the zinc supplied; two reasons at least could be pointed out to explain such a fact: 1 - Zinc fixation by an exchange with magnesium or by filling holes in the octahedral layer of aluminosilicates, probably kaolinite; 2 - No need for fertilizer zinc in the particular stage of life cycle under which the experiment was set up. The data from chemical analysis are roughly parallel to the above mentioned. When one attempts to compare - by taking data herein reported zinc uptake from nutrient solution, leaf brushing or from fertilizers in the soil, a practical conclusion can be drawn: the control of zinc deficiency in coffee plants should not be done by adding the zinc salts to the soil; in other words: the soil applications used so extensively in other countries seem not to be suitable for our conditions; hence zinc sprays should be used wherever necessary.

Estudo sobre a Alimentação Mineral do algodoeiro I: marcha da absorpção do macronutrientes (Nota prévia)

Cotton (variety I. A. C. 11) was grown on a sandy soil under two treatments, namely: (1) NPK + lime and (2) no fertilizers. Three weeks after planting a systematic sampling of entire plants was done every other week. In the laboratory determinations of dry weight were made and afterwards the various plant partes were submitted to chemical analyses, nitrogen (N), phosphorus (P), potassium (K), calcium (Ca), magnesium (Mg), and sulfur (S) being determined. The aim of this work was to obtain information on the periods in which the absorption of the several macronutrients was more intense, this providing a clue for time of application of certain mineral fertilizers. Data obtained hereby allowed for the following main conclusions. The initial rate of growth of the cotton plant, judged by the determinations of dry weight, is rather slow. Seven weeks after planting and again five weeks two distinct periods of rapid growth take place. The uptake of macronutrients is rather small until the first flowers show up. From there on the absorption of minerals is intensified. From the time in which fruits are being formed to full maturity, the crop draws from the soil nearly 75 percent of the total amount of elements required to complet life cycle. This seams to point out the need for late dressings of fertilizers, particularly of those containing N and K. The following amounts of element in Kg/ha were absorbed by the fertilized plants: N - 83.2 P - 8.1 K - 65.5 Ca - 61.7 Mg - 12.8 and S - 33.2. The three major macronutrients, namely, N. P and K are exported as seed cotton in the following proportions with respect to the total amounts taken up by the entire crop: N - 1/3, P - 1/2 and K - 1/3.

Nutrient uptake and nutritional efficiency in sweet sorghum

Two sweet sorghum varieties, Brandes and Rio, were grown in full strenght and diluted nutrient solutions till completing the life cycle wherein mineral analyses were carried out. As a rule both varieties showed the same capacity to absorb nutrients in the two rates supplied. Dry matter yield, however was different in the dilute nutrient solution. The variety Brandes produced more fresh stalks in the full strength solution than Rio; under nutricional stress the yield was lower. Dry matter of stalks in the case of the variety Rio was consistently higher.

Aspects of the biology and behavior of Nesolynx sp. (Hymenoptera, Eulophidae)

This research was carried out to study some aspects of the biology and behavior of Nesolynx sp. (Hymenoptera, Eulophidae), a pupal parasite of Psorocampa denticulata (Lepidoptera, Notodontidae) a defoliating caterpillar of Eucalyptus spp. in Brazil. The adults emerge from the host pupa through a circular hole on Its dorsal region. Mating occurs righ after the emergence and the longevity of adults was two days for the males and four days for the females. Regarding to the host species Diatraea saccharalis showed a number of adults significantly greater than Galleria mellonella and the increasing temperature from 21±1 °C to 26±1°C caused a significative increasing in the number of emerged adults in both host species. The emergence of adults increased proportionally to the period of exposition to the host up to 3.50 days; after that, a considerable decrease in the emergence was observed. The parasitoid showed parthenogenetic reproduction therefore the average number of emerged males was significantly greater than the number of females. The sex ratio was similar for the insects emerged from virgin or mated females (0,96) and the life cycle lenght was around 18.34 days for both conditions.

Relacionamento entre anfíbios anuros e bromélias da restinga de Regência, Linhares, Espírito Santo, Brasil

Some anuran amphibians use the bromeliads during the entire life cycle and others only as diurnal shelter. At the sandy coastal plain of Linhares, State of Espírito Santo, Southeastern Brazil, 676 bromeliads were examined, of which 303 of Aechmea blanchetiana (Baker) L. B. Smith., 1955, 287 of Aechmea nudicaulis (L.) Griseb., 1864, and 86 of Vrisea procera (Mart. Ex Schult. f.) Wittm, 1891. The morphometric and physical-chemical analysis of different bromeliads evidenced variations among plants. During the period sampled, six anuran species were found inside the plant axils. The hylid frog Phyllodytes luteolus (Wied, 1824) was the most abundant species (260 specimens). Its abundance was higher in the epiphyte bromeliad Vrisea procera. Phyllodytes luteolus had higher occurrences in bromeliads located at a transitional area between open and under the shrub vegetation. Specimens of Scinax alterus (Lutz, B., 1973) and Aparasphenodon brunoi Miranda-Ribeiro, 1920, were more frequent mainly in transitional areas; Bufo granulosus Spix, 1824 occurred in open and transitional areas, whereas Gastrotheca fissipis (Boulenger, 1888) and Leptodactylus fuscus (Schneider, 1799) were found only in bromeliads located in open areas.

Fish assemblage composition in a tributary of the Mogi Guaçu river basin, southeastern Brazil

Fish assemblage composition and seasonal patterns of species abundance were studied in Cabaceiras stream, a tributary of the Mogi Guaçu river in São Paulo State, Brazil. Three stations were sampled monthly from June 1999 to May 2000 using sieves and small trawl net and gill nets. Fifteen fish families, 37 genera and 45 species were captured. Characiformes (27 spp.) and Siluriformes (13 spp.) were the most species-rich orders. Gymnotiformes and Perciformes were represented by two species each, and Synbranchiformes had only a single species. One group of species (approximately 75 %) persisted in the stream throughout the year. A second group (approximately 25 %) contained species that only occupy the stream for a limited period of their life cycle, and overall fish assemblage composition was associated with the seasonal flood cycle.

Exigências térmicas para o desenvolvimento de Palmistichus elaeisis (Hymenoptera, Eulophidae) em pupas de cinco espécies de lepidópteros

The thermal requirements of Palmistichus elaeisis Delvare & LaSalle, 1993 were determined by rearing this pupal parasitoid in the following lepidopteran hosts: Diatraea saccharalis (Fabricius, 1794) (Crambidae), Anticarsia gemmatalis (Hübner, 1818), Heliothis virescens (Fabricius, 1777), Spodoptera frugiperda (J.E. Smith, 1797) (Noctuidae) and Thyrinteina arnobia (Stoll, 1782) (Geometridae). The experiment was set in laboratory conditions (temperatures between 18ºC and 30ºC, RH=70 ± 10% and 14-hour photoperiod). The life cycle (egg-adult) ranged from 40.4 ± 0.63 days (18ºC, H. virescens) to 18.1 ± 0.13 days (28ºC, D. saccharalis). The thermal constant of the egg-adult period ranged from 353.1 (D. saccharalis) to 407.7 degree-days (S. frugiperda). The lowest thermal threshold ranged from 5.0ºC (A. gemmatalis) to 7.5ºC (D. saccharalis). At 30ºC, all prepupae died. These results suggest that P. elaeisis can be reared in laboratory on all the hosts tested.

The first record of Litargus tetraspilotus (Coleoptera, Mycetophagidae) in Brazil, with biological notes and complementary description of the species

Litargus tetraspilotus LeConte, 1856 was collected feeding on Oidium sp. (Fungi, Ascomycota, Erysiphaceae) associated with fruit trees. This is the first time L. tetraspilotus is recorded in Brazil, totaling three species of Mycetophagidae for this country. This study aims to provide a complementary description of this species based on new characters and to present information on its life cycle under laboratory conditions and fluctuation in population in the field. During the period of inventories between July 2004 and August 2006, about every fifteen days, a total of 565 specimens of L. tetraspilotus were collected, with the highest abundance found on citrus plants, with values differing significantly between the two years. The population levels differed between the seasons; spring had the greatest abundance and autumn the least. There was a significant positive correlation of L. tetraspilotus abundance with rainfall and relative humidity. Mycetophagidae, as well as other mycophagous families of Brazilian coleopterans, are barely studied, warranting further future studies of their bioecology and systematics.

Compared nymphal development of Tingis americana (Hemiptera, Tingidae) in two Handroanthus species (Bignoniaceae) and reproductive parameters in seedlings of Handroanthus heptaphyllus

Nymphs and adults of Tingis americana Drake, 1922 were found feeding on leaves of Handroanthus heptaphyllus (Vell.) Mattos and Handroanthus chrysotrichus (Mart. ex A. DC.) Mattos in the Botanic Garden, Porto Alegre, state of Rio Grande do Sul, Brazil. This is the first record of T. americana on these host plants and in the southern Brazil. We aimed to compare the nymphal development on both hosts and to analyze the reproductive parameters on H. heptaphyllus (25 ± 1ºC; 60 ± 10% RH; 16 h photophase). The mean nymphal period (days) was shorter in individuals reared on H. heptaphyllus (12.69 ± 0.076) than on H. chrysotrichus (19.11 ± 0.208) (P < 0.0001), however, nymph viability was similar. On H. heptaphyllus, the mean embryonic period lasted 12.32 ± 0.274 days and the egg viability was 92%. The mean total and daily fecundity were 310.0 ± 19.40 eggs/female and 7.46 ± 0.302 eggs/female/day, respectively. Paired males and females showed similar longevity (P = 0.0691), while unpaired females lived longer than unpaired males (P = 0.0460).

Genome of an arbuscular mycorrhizal fungus provides insight into the oldest plant symbiosis.

The mutualistic symbiosis involving Glomeromycota, a distinctive phylum of early diverging Fungi, is widely hypothesized to have promoted the evolution of land plants during the middle Paleozoic. These arbuscular mycorrhizal fungi (AMF) perform vital functions in the phosphorus cycle that are fundamental to sustainable crop plant productivity. The unusual biological features of AMF have long fascinated evolutionary biologists. The coenocytic hyphae host a community of hundreds of nuclei and reproduce clonally through large multinucleated spores. It has been suggested that the AMF maintain a stable assemblage of several different genomes during the life cycle, but this genomic organization has been questioned. Here we introduce the 153-Mb haploid genome of Rhizophagus irregularis and its repertoire of 28,232 genes. The observed low level of genome polymorphism (0.43 SNP per kb) is not consistent with the occurrence of multiple, highly diverged genomes. The expansion of mating-related genes suggests the existence of cryptic sex-related processes. A comparison of gene categories confirms that R. irregularis is close to the Mucoromycotina. The AMF obligate biotrophy is not explained by genome erosion or any related loss of metabolic complexity in central metabolism, but is marked by a lack of genes encoding plant cell wall-degrading enzymes and of genes involved in toxin and thiamine synthesis. A battery of mycorrhiza-induced secreted proteins is expressed in symbiotic tissues. The present comprehensive repertoire of R. irregularis genes provides a basis for future research on symbiosis-related mechanisms in Glomeromycota.