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

Trypanosoma cruzi-elicited CD8+ T cell-mediated myocarditis: chemokine receptors and adhesion molecules as potential therapeutic targets to control chronic inflammation?

In Chagas disease, during the acute phase, the establishment of inflammatory processes is crucial for Trypanosoma cruzi control in target tissues and for the establishment of host/parasite equilibrium. However, in about 30% of the patients, inflammation becomes progressive, resulting in chronic disease, mainly characterized by myocarditis. Although several hypothesis have been raised to explain the pathogenesis of chagasic myocardiopathy, including the persistence of the parasite and/or participation of autoimmune processes, the molecular mechanisms underlying the establishment of the inflammatory process leading to parasitism control but also contributing to the maintenance of T. cruzi-elicited chronic myocarditis remain unsolved. Trying to shed light on these questions, we have for several years been working with murine models for Chagas disease that reproduce the acute self-resolving meningoencephalitis, the encephalitis resulting of reactivation described in immunodeficient individuals, and several aspects of the acute and chronic myocarditis. In the present review, our results are summarized and discussed under the light of the current literature. Furthermore, rational therapeutic intervention strategies based on integrin-mediated adhesion and chemokine receptor-driven recruitment of leukocytes are proposed to control T. cruzi-elicited unbalanced inflammation.

A new serovar and a new serological variant belonging to Salmonella enterica subspecies diarizonae

Description of a new serovar (S. IIIb 16:k:e,n,x,z15) and a new serological variant (S. IIIb 42:z10:e,n,x,z15:z60 ) belonging to the genus Salmonella isolated from stool specimens of Brazilian snakes (Crotalus durissus).

Cross-immunoreactivity between anti-potato apyrase antibodies and mammalian ATP diphosphohydrolases: potential use of the vegetal protein in experimental schistosomiasis

We have previously showed that Schistosoma mansoni ATP-diphosphohydrolase and Solanum tuberosum potato apyrase share epitopes and the vegetable protein has immunostimulatory properties. Here, it was verified the in situ cross-immunoreactivity between mice NTPDases and anti-potato apyrase antibodies produced in rabbits, using confocal microscopy. Liver samples were taken from Swiss Webster mouse 8 weeks after infection with S. mansoni cercariae, and anti-potato apyrase and TRITC-conjugated anti-rabbit IgG antibody were tested on cryostat sections. The results showed that S. mansoni egg ATP diphosphohydrolase isoforms, developed by anti-potato apyrase, are expressed in miracidial and egg structures, and not in granulomatous cells and hepatic structures (hepatocytes, bile ducts, and blood vessels). Therefore, purified potato apyrase when inoculated in rabbit generates polyclonal sera containing anti-apyrase antibodies that are capable of recognizing specifically S. mansoni ATP diphosphohydrolase epitopes, but not proteins from mammalian tissues, suggesting that autoantibodies are not induced during potato apyrase immunization. A phylogenetic tree obtained for the NTPDase family showed that potato apyrase had lower homology with mammalian NTPDases 1-4, 7, and 8. Further analysis of potato apyrase epitopes could implement their potential use in schistosomiasis experimental models.

Parasitism of two zoonotic reservoirs Dasyprocta leporina and D. fuliginosa (Rodentia) from Amazonas, with Trichostrongylina nematodes (Heligmonellidae): description of a new genus and a new species

A new genus and a new species of Heligmonellidae nematodes are described parasiting the stomach of three agoutis (two Dasyprocta fuliginosa and one D. leporina) captured in the middle and high Negro river microregion, state of Amazonas, Brazil. The new genus, as well as its type-species, are closely related to the trichostrongylids included in Fuellebornema, particularly on what concerns the pattern of the caudal bursa, but differing from them by the characteristics of the synlophe, that presents a poorly developed carene, when compared to the referred number of body ridges in Freitastrongylus n. gen. and consequently in F. angelae n. sp.,in which the ridges are well developed and the carene at mid-body has a similar size when compared to the ridge situated in front of the right field (ridge no. 5). Caudal bursa is of the type 1-4, with rays 9 shorter than rays 10, with a very long genital cone.

Resurrection of Anopheles goeldii from synonymy with Anopheles nuneztovari (Diptera, Culicidae) and a new record for Anopheles dunhami in the Brazilian Amazon

Nucleotide sequences of the internal transcribed spacer 2 (ITS2) rDNA and partial sequences of the cytochrome coxidase subunit I (COI) mtDNA and white gene nDNA were obtained from specimens of Anopheles nuneztovari A collected in Macapá (state of Amapá), Óbidos, Prainha and Almeirim (state of Pará), Itacoatiara and Parintins (state of Amazonas), Brazil, and compared with previously published sequences of A. nuneztovari s.l. Results of the Bayesian phylogenetic analyses performed using either COI or combined ITS2, COI and white gene sequences suggest that An. nuneztovari B/C is distinct from specimens obtained in the Amazonas/Solimões River basin. Anopheles goeldii, currently in synonymy with An. nuneztovari, was described from individuals collected in Belterra (= Fordlândia) in the Tapajós River, state of Pará, Southern Amazonas River. Morphological comparisons of the characteristics of the male genitalia indicated that An. nuneztovari A and An. goeldii are similar but distinct from An. nuneztovariB/C by the apex of the aedeagus. In considering the results of the phylogenetic analyses and morphological comparisons, An. goeldii is resurrected from synonymy with An. nuneztovari. Additionally, Anopheles dunhamiis reported for the first time in Parintins. This species can be distinguished from An. goeldiiby characters of the male genitalia and molecular data.

Ecoepidemiology, short history and control of Chagas disease in the endemic countries and the new challenge for non-endemic countries

Chagas disease is maintained in nature through the interchange of three cycles: the wild, peridomestic and domestic cycles. The wild cycle, which is enzootic, has existed for millions of years maintained between triatomines and wild mammals. Human infection was only detected in mummies from 4,000-9,000 years ago, before the discovery of the disease by Carlos Chagas in 1909. With the beginning of deforestation in the Americas, two-three centuries ago for the expansion of agriculture and livestock rearing, wild mammals, which had been the food source for triatomines, were removed and new food sources started to appear in peridomestic areas: chicken coops, corrals and pigsties. Some accidental human cases could also have occurred prior to the triatomines in peridomestic areas. Thus, triatomines progressively penetrated households and formed the domestic cycle of Chagas disease. A new epidemiological, economic and social problem has been created through the globalisation of Chagas disease, due to legal and illegal migration of individuals infected by Trypanosoma cruzi or presenting Chagas disease in its varied clinical forms, from endemic countries in Latin America to non-endemic countries in North America, Europe, Asia and Oceania, particularly to the United States of America and Spain. The main objective of the present paper was to present a general view of the interchanges between the wild, peridomestic and domestic cycles of the disease, the development of T. cruzi among triatomine, their domiciliation and control initiatives, the characteristics of the disease in countries in the Americas and the problem of migration to non-endemic countries.

New classification of natural breeding habitats for Neotropical anophelines in the Yanomami Indian Reserve, Amazon Region, Brazil and a new larval sampling methodology

Here we present the first in a series of articles about the ecology of immature stages of anophelines in the Brazilian Yanomami area. We propose a new larval habitat classification and a new larval sampling methodology. We also report some preliminary results illustrating the applicability of the methodology based on data collected in the Brazilian Amazon rainforest in a longitudinal study of two remote Yanomami communities, Parafuri and Toototobi. In these areas, we mapped and classified 112 natural breeding habitats located in low-order river systems based on their association with river flood pulses, seasonality and exposure to sun. Our classification rendered seven types of larval habitats: lakes associated with the river, which are subdivided into oxbow lakes and nonoxbow lakes, flooded areas associated with the river, flooded areas not associated with the river, rainfall pools, small forest streams, medium forest streams and rivers. The methodology for larval sampling was based on the accurate quantification of the effective breeding area, taking into account the area of the perimeter and subtypes of microenvironments present per larval habitat type using a laser range finder and a small portable inflatable boat. The new classification and new sampling methodology proposed herein may be useful in vector control programs.

A new genus and nine new species of Neotropical spittlebugs (Hemiptera, Cercopidae, Tomaspidinae)

The following new taxa of South American Cercopidae are described: Ferorhinella gen. nov., with type species Sphenorhina brevis Walker, 1851; Deois (Deois) knighti sp. nov. (Brazil), Deois (Deois) sexpunctata sp. nov. (Brazil), Deois (Pandysia) constricta sp. nov. (Brazil), Mahanarva (Ipiranga) bahiaensis sp. nov. (Brazil), Maxantonia bifurcata sp. nov. (Brazil), Neosphenorhina curvipenis sp. nov. (Brazil), Sphenorhina brevispina sp. nov. (Ecuador), Sphenorhina nigricephala sp. nov. (Ecuador), Sphenorhina minuta sp. nov. (Brazil), Tropidorhinella onorei sp. nov. (Colombia), Zuata luteofascia sp. nov. (Colombia). Ferorhinella brevis (Walker, 1851) comb. nov. In addition, descriptions are given for a new colour form of Mahanarva (M.) phantastica (Breddin, 1904) and a newly found variation in the male genitalia of Deoisella fasciata Costa & Sakakibara, 2002.

Survey of ear flies (Diptera, Ulidiidae) in maize (Zea mays L.) and a new record of Euxesta mazorca Steyskal in Brazil

Survey of ear flies (Diptera, Ulidiidae) in maize (Zea mays L.) and a new record of Euxesta mazorca Steyskalin Brazil. Species of Euxesta (Diptera, Ulidiidae), known as silk flies or ear flies, are becoming increasingly important as maize insect pests in South America, although very little is known about them in Brazil. The larvae of some species of this genus initially damage female reproductive tissues, and then the developing kernels on the ear. As a result of feeding, fermentation and associated odors cause complete loss of the grain because it is no longer fit for human or livestock consumption. The main objective of this work was to evaluate the incidence of Euxesta spp. in Brazilian maize fields and to determine the most prevalent species using two different hydrolyzed protein foods attractants, BioAnastrepha® (hydrolyzed maize protein) and Torula, placed inside McPhail traps. The two species identified were E. eluta Loew and E. mazorca Steyskal, the latter being a new record from Brazil. Between the two species, E. eluta was the more abundant in maize fields. Both attractants were efficient in capturing the two species. However, BioAnastrepha® captured significantly more insects than Torula.

Sand flies (Diptera, Psychodidae, Phlebotominae) from Central Amazonia and four new records for the Amazonas state, Brazil

Sand flies from Central Amazonia and four new records for the Amazonas state, Brazil. A survey was conducted in May and June 2008 to study the fauna of insects in Central Amazonia, Brazil. As part of the survey, we report here that sixty species of phlebotomine were identified, totaling 13,712 specimens from 13 genera. The collection sites were located at the border between the states of Pará and Amazonas, comprising three municipalities from the Amazonas state (Borba, Maués, and Nhamundá). Malaise, CDC and Shannon traps were used to collect the insects. Most of the sand flies were collected by CDC traps (89.5%), while Malaise and Shannon traps collected 7% and 3.5%, respectively. The most abundant genera, representing 97.1% of the total sand flies identified were: Trichopygomyia Barretto, 1962 (47.6%), Psathyromyia Barretto, 1962 (17.9%), Psychodopygus Mangabeira, 1941 (17.5%) and Trichophoromyia Barretto, 1962 (14.3%). The genera with the largest number of species identified were: Psychodopygus (14), Psathyromyia (10), Evandromyia Mangabeira, 1941 (7), Trichophoromyia (5) and Trichopygomyia (5). The most abundant species was Trichopygomyia trichopyga (Floch & Abonnenc, 1945), which represented 29% of the total sand flies identified. Here we also report new records for four species in the Amazonas state: Ps. complexus (Mangabeira, 1941), Ps. llanosmartinsi Fraiha & Ward, 1980, Ty. pinna (Feliciangeli, Ramirez-Pérez & Ramirez, 1989), and Th. readyi (Ryan, 1986). The results of this study provide new, additional information on the distribution of sand flies in the Amazon and increase the number of species in the Amazonas state from 127 to 131.

Distribution and agroclimatic characterization of potential cultivation regions of physic nut in Mexico

The purposes of this study were to determine the distribution and climatic patterns of current and future physic nut (Jatropha curcas) cultivation regions in Mexico, and to identify possible locations for in vivo germplasm banks establishment, using geographic information systems. Current climatic data were processed by Floramap software to obtain distribution maps and climatic patterns of regions where wild physic nuts could be found. DIVA-GIS software analyzed current climatic data (Worldclim model) and climatic data generated by CCM3 model to identify current and future physic nut cultivation regions, respectively. The distribution map showed that physic nut was present in most of the tropical and subtropical areas of Mexico, which corresponded to three agroclimatic regions. Climate types were Aw2, Aw1, and Bs1, for regions 1, 2 and 3, respectively. Nontoxic genotypes were associated with region 2, and toxic genotypes were associated with regions 1 and 3. According to the current and future cultivation regions identified, the best suitable ones to establish in vivo germplasm collections were the coast of Michoacán and the Isthmus of Tehuantepec, located among the states of Veracruz, Oaxaca and Chiapas.

Stocking densities and feeding strategies in shrimp and tilapia polyculture in tanks

The objective of this work was to evaluate the performance of Pacific marine shrimp (Litopenaeus vannamei) and tilapia (Oreochromis niloticus), in a polyculture in tanks subjected to different stocking densities and feeding strategies, in comparison with monoculture. Two experiments were performed, at the same time, in a completely randomized design with three treatments and four replicates each. Treatments for experiment I were: monoculture with 10 shrimp per m² (10S:0T); polyculture with 10 shrimp and 0.5 tilapia per m² (10S:0.5T); and polyculture with 10 shrimp and 1 tilapia per m² (10S:1T). Shrimp was the main crop, and feed was provided based on shrimp biomass. Treatments for experiment II were: monoculture with 2 tilapia per m² (2T:0S); polyculture with 2 tilapia and 2.5 shrimp per m² (2T:2.5S); and polyculture with 2 tilapia and 5 shrimp per m² (2T:5S). Tilapia was the main crop, and feed was provided based on fish requirements. In the experiment I, tilapia introduction to shrimp culture resulted in lower shrimp growth and poor feed conversion rate. In experiment II, shrimp introduction to tilapia culture did not interfere with fish performance. Polyculture is more efficient with the combination of 2 tilapia and 2.5 or 5 shrimp per m² and feed based on fish requirements.

CHARACTERIZATION AND EVALUATION OF THE POTENTIAL USE OF SLUDGE FROM STP AND WTP IN PAVING

ABSTRACT Water and sewage treatment plants (STP and WTP) generate as byproduct a significant amount of sludge with environment harmful elements. Sending to landfills or depositing on the ground or rivers are respectively expensive and dangerous alternatives. In this scenario, the use of this waste in paving processes is a promising alternative for disposal thereof. In this study, we focused on characterizing sludge and evaluating its use in paving, which showed satisfactory results for use in base and sub-base floors.

Comparison of nuclear grade and immunohistochemical features in situ and invasive components of ductal carcinoma of breast

PURPOSE:To compare the prognostic and predictive features between in situ and invasive components of ductal breast carcinomas. METHODS:We selected 146 consecutive breast samples with ductal carcinoma in situ (DCIS) associated with adjacent invasive breast carcinoma (IBC). We evaluated nuclear grade and immunohistochemical expression of estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2 (HER2), cytokeratin 5/6 (CK5/6), and epidermal growth factor receptor (EGFR) in both components, in situ and invasive, and the Ki-67 percentage of cells in the invasive part. The DCIS and IBC were classified in molecular surrogate types determined by the immunohistochemical profile as luminal (RE/PR-positive/ HER2-negative), triple-positive (RE/RP/HER2-positive), HER2-enriched (ER/PR-negative/HER2-positive), and triple-negative (RE/RP/HER2-negative). Discrimination between luminal A and luminal B was not performed due to statistical purposes. Correlations between the categories in the two groups were made using the Spearman correlation method. RESULTS:There was a significant correlation between nuclear grade (p<0.0001), expression of RE/RP (p<0.0001), overexpression of HER2 (p<0.0001), expression of EGFR (p<0.0001), and molecular profile (p<0.0001) between components in situ and IBC. CK 5/6 showed different distribution in DCIS and IBC, presenting a significant association with the triple-negative phenotype in IBC, but a negative association among DCIS. CONCLUSIONS: Our results suggest that classical prognostic and predictive features of IBC are already determined in the preinvasive stage of the disease. However the role of CK5/6 in invasive carcinoma may be different from the precursor lesions.