Experimental Infection of Calomys callosus (Rodentia, Cricetidae) by Toxoplasma gondii

Calomys callosus, Rengger 1830 (Rodentia, Cricetidae), a wild rodent found in Central Brazil, was studied to investigate its susceptibility to Toxoplasma gondii experimental infection and its humoral immune response against this protozoa. The electrophoretic profile of the serum proteins of C. callosus showed that IgG, which shows no affinity to Protein A, has higher cross reactivity with rat IgG than with IgG from other rodents. The susceptibility assay was performed by inoculation groups of animals with various suspensions of T. gondii tachyzoites from 102 to 106 parasites. All animals died between 3 and 9 days after infection and the kinetics of antibody synthesis was determined. Basically, they recognized predominantly the immunodominant antigen SAG-1 (P30). The immunohistochemistry assays revealed that the liver was the most heavily infected organ, followed by the spleen, lungs, intestine, brain and kidneys. It can be concluded that C. callosus is an excellent experimental model for acute phase of Toxoplasma infection

Differential gene expression during Trypanosoma cruzi metacyclogenesis

The transformation of epimastigotes into metacyclic trypomastigotes involves changes in the pattern of expressed genes, resulting in important morphological and functional differences between these developmental forms of Trypanosoma cruzi. In order to identify and characterize genes involved in triggering the metacyclogenesis process and in conferring to metacyclic trypomastigotes their stage specific biological properties, we have developed a method allowing the isolation of genes specifically expressed when comparing two close related cell populations (representation of differential expression or RDE). The method is based on the PCR amplification of gene sequences selected by hybridizing and subtracting the populations in such a way that after some cycles of hybridization-amplification genes specific to a given population are highly enriched. The use of this method in the analysis of differential gene expression during T. cruzi metacyclogenesis (6 hr and 24 hr of differentiation and metacyclic trypomastigotes) resulted in the isolation of several clones from each time point. Northern blot analysis showed that some genes are transiently expressed (6 hr and 24 hr differentiating cells), while others are present in differentiating cells and in metacyclic trypomastigotes. Nucleotide sequencing of six clones characterized so far showed that they do not display any homology to gene sequences available in the GeneBank.

Nematodes of elasmobranch fishes from the southern coast of Brazil

New records for nematode species recovered from elasmobranch fishes in Brazil are established and new systematical arrangements proposed. Parascarophis sphyrnae Campana-Rouget, 1955 from the spiral valve of Sphyrna zygaena is referred for the first time in South America as a new host record. Procamallanus (S.) pereirai Annereaux, 1946, from the spiral valve of Raja castelnaui is reported parasitizing an elasmobranch host. Nematode larvae of the genera Anisakis, Contracaecum, Pseudoterranova and Raphidascaris are listed from the stomach and spiral valves of several hosts. Anisakidae larvae previously referred in Brazil in the genus Phocanema should be reallocated in Pseudoterranova. Nematodes of the genera Anisakis, Contracaecum, Pseudoterranova and Raphidascaris are reported for the first time parasitizing elasmobranchs in Brazil.

Digenea and acanthocephala of elasmobranch fishes from the southern coast of Brazil

New records for helminth species recovered from elasmobranch fishes in Brazil are established. Digenean and acanthocephalan parasites of elasmobranch fishes are reported from the southern coast of Brazil: Otodistomum veliporum (Creplin, 1837) Stafford, 1904 (Digenea: Azygiidae) in the stomach and spiral valve of Dipturus trachydermus and in the spiral valve of Squatina sp. Cystacanths and juveniles of the acanthocephalans Corynosoma australe Johnston, 1937 and Corynosoma sp., in the spiral valve of Squatina sp., Galeorhinus galeus and Hexanchus griseus and in the stomach of Squalus megalops; a juvenile of Gorgorhynchus sp., in the spiral valve of Sphyrna zygaena. Dipturus trachydermus and Squatina sp. are new host records for O. veliporum. Digeneans and acanthocephalans are reported for the first time parasitizing elasmobranch fishes in Brazil.

Taxonomic reports of Otobothrioidea (Eucestoda, Trypanorhyncha) from elasmobranch fishes of the southern coast off Brazil

Specimens of elasmobranch fishes, captured in the states of Paraná and Santa Catarina, of the southern coast off Brazil, represented by three families, four genera, and four species, were parasitized with otobothrioid trypanorhynch cestodes: Heptranchias perlo (Bonnaterre, 1788), Squalus sp. and Carcharhinus signatus (Poey, 1868) were parasitized with Progrillotia dollfusi Carvajal & Rego,1987; Prionace glauca (Linnaeus, 1758) with Molicola horridus (Goodsir, 1841) Dollfus, 1942. Details of internal morphology and/or scolex and/or proglottids surface ultrastructure, that expanded the description of M. horridus, through observations with lightfield, and/or scanning eletronic microscopy, are provided. The known geographical distribution for the species M. horridus is enlarged. P. dollfusi is reported for the first time in elasmobranchs.

Trypanosoma cruzi strains isolated from human, vector, and animal reservoir in the same endemic region in Mexico and typed as T. cruzi I, discrete typing unit 1 exhibit considerable biological diversity

In this study, three strains of Trypanosoma cruzi were isolated at the same time and in the same endemic region in Mexico from a human patient with chronic chagasic cardiomyopathy (RyC-H); vector (Triatoma barberi) (RyC-V); and rodent reservoir (Peromyscus peromyscus) (RyC-R). The three strains were characterized by multilocus enzyme electrophoresis, random amplified polymorphic DNA, and by pathological profiles in experimental animals (biodemes). Based on the analysis of genetic markers the three parasite strains were typed as belonging to T. cruzi I major group, discrete typing unit 1. The pathological profile of RyC-H and RyC-V strains indicated medium virulence and low mortality and, accordingly, the strains should be considered as belonging to biodeme Type III. On the other hand, the parasites from RyC-R strain induced more severe inflammatory processes and high mortality (> 40%) and were considered as belonging to biodeme Type II. The relationship between genotypes and biological characteristics in T. cruzi strains is still debated and not clearly understood. An expert committee recommended in 1999 that Biodeme Type III would correspond to T. cruzi I group, whereas Biodeme Type II, to T. cruzi II group. Our findings suggest that, at least for Mexican isolates, this correlation does not stand and that biological characteristics such as pathogenicity and virulence could be determined by factors different from those identified in the genotypic characterization

Clinical forms of Trypanosoma cruzi infected individuals in the chronic phase of Chagas disease in Puebla, Mexico

In Mexico, despite the relatively high seroprevalence of Trypanosoma cruzi infection in humans in some areas, reported morbidity of Chagas disease is not clear. We determined clinical stage in 71 individuals seropositive to T. cruzi in the state of Puebla, Mexico, an area endemic for Chagas disease with a reported seroprevalence of 7.7%. Diagnosis of Chagas disease was made by two standardized serological tests (ELISA, IHA). Individuals were stratified according to clinical studies. All patients were submitted to EKG, barium swallow, and barium enema. Groups were identified as indeterminate form (IF) asymptomatic individuals without evidence of abnormalities (n = 34 cases); those with gastrointestinal alterations (12 patients) including symptoms of abnormal relaxation of the lower esophageal sphincter and absent peristalsis in the esophageal body, grade I megaesophagus, and/or megacolon; patients with clinical manifestations and documented changes of chronic Chagas heart disease who were subdivided as follows: mild (8 patients) - mild electrocardiographic changes of ventricular repolarization, sinus bradychardia); moderate (6 patients) - left bundle branch block, right bundle branch block associated with left anterior fascicular block); severe (8 patients) - signs of cardiomegaly, dilated cardiomyopathy); and the associated form (3 cases) that included presence of both cardiomyopathy and megaesophagus. These data highlight the importance of accurate evaluation of the prevalence and clinical course of Chagas disease in endemic and non-endemic areas of Mexico.

Myxidium volitans sp. nov., a parasite of the gallbladder of the fish, Dactylopterus volitans (Teleostei: Triglidae) from the Brazilian Atlantic coast: morphology and pathology

Myxidium volitans sp. nov. (Myxozoa: Myxidiidae) parasitizing the hypertrophied green-brownish gallbladder of the teleost Dactylopterus volitans, collected in the Atlantic coast near Niterói, Brazil was described based on ultrastructural studies. The spores were fusiform, sometimes slightly crescent-shaped on average 21.7 ± 0.3 µm (mean ± standard deviation) (n = 50) long and 5.6 ± 0.4 µm (n = 30) wide. The spore wall was thin and smooth, comprising two equally-sized valves joined by a hardly visible sutural ridge. Spores containing two pyriform polar capsules (PC) (5.0 ± 0.4 × 2.3 ± 0.3 µm) (n = 30) are situated in each extremity of the spore. The PC wall was composed of hyaline layer (0.20-0.29 µm thick) and by a thin external granular layer. Each PC contains a polar filament (PF) with irregular arrangements that was projected from its apical region to the bases of PC and coiled laterally from bases to the tip of PC. Some regular striations and S-like structures in the periphery of the PFs with four-five irregular sections were observed. Based on the spore morphology, ultrastructural differences and the specificity of the host we describe this parasite as a new myxosporidian, named M. volitans sp. nov.

Ultrastructural description of Ceratomyxa microlepis sp. nov. (Phylum Myxozoa): a parasite infecting the gall bladder of Hemiodus microlepis, a freshwater teleost from the Amazon River

A new ceratomyxid parasite was examined for taxonomic identification, upon being found infecting the gall bladder of Hemiodus microlepis (Teleostei: Hemiodontidae), a freshwater teleost collected from the Amazon River, Brazil. Light and transmission electron microscopy revealed elongated crescent-shaped spores constituted by two asymmetrical shell valves united along a straight sutural line, each possessing a lateral projection. The spores body measured 5.2 ± 0.4 µm (n = 25) in length and 35.5 ± 0.9 µm (n = 25) in total thickness. The lateral projections were asymmetric, one measuring 18.1 ± 0.5 µm (n = 25) in thickness and the other measuring 17.5 ± 0.5 µm (n = 25) in thickness. Two equal-sized subspherical polar capsules measuring 2.2 ± 0.3 µm in diameter were located at the same level, each possessing a polar filament with 5-6 coils. The sporoplasm was binucleate. Considering the morphometric data analyzed from the microscopic observations, as well as the host species and its geographical location, this paper describes a new myxosporean species, herein named Ceratomyxa microlepis sp. nov.; therefore representing the first description of a freshwater ceratomyxid from the South American region.

O impacto das novas mídias para os anunciantes brasileiros

O surgimento das tecnologias digitais permitiu o desenvolvimento de meios de contato inéditos entre empresas e consumidores, as chamadas novas mídias. Capazes de reconfigurar indústrias como a fonográfica e a do cinema, as novas mídias têm também impacto considerável na indústria da publicidade. O subsídio cruzado entre anunciantes, veículos de massa e consumidores, consolidado ao longo do século XX, se vê fragilizado com a popularização de aparelhos como Digital Video Recorders (DVRs), tablets, consoles de videogames e telefones celulares, dispositivos que transferem aos consumidores o poder de regular sua exposição à publicidade. Nesse novo ambiente, mídias tradicionais e novas mídias competem pela preferência de consumidores e pela verba dos anunciantes. Apesar da necessidade dos anunciantes de aferir e rentabilizar campanhas publicitárias, verificou-se que o investimento em veículos tradicionais vem crescendo, enquanto a audiência entregue diminui. Num panorama de mudanças, no presente estudo procura-se identificar que fatores influenciam o processo de compra de mídias por parte dos anunciantes, observando a percepção deles sobre mídias emergentes e tradicionais, seus benefícios e desafios. Para tanto, foi realizada uma pesquisa exploratória, utilizando uma abordagem qualitativa. Entre junho e julho de 2009, foram realizadas entrevistas em profundidade com os profissionais responsáveis pelas campanhas publicitárias de 15 empresas anunciantes de grande porte, atuantes em diversas indústrias. Ao analisar os dados colhidos em campo à luz da literatura estudada sobre o tema, a pesquisa identificou três fatores com forte influência sobre a definição do mix de mídias dos entrevistados: o uso de métricas inadequadas às mídias emergentes, a dependência dos anunciantes com as agências de publicidade e os veículos de massa e os próprios processos internos dos anunciantes.

Validity and reliability of a questionnaire on knowledge of cardiovascular risk factors for use in Brazil

Using a sample of patients with coronary artery disease, this methodological study aimed to conduct a cross-cultural adaptation and validation of a questionnaire on knowledge of cardiovascular risk factors (Q-FARCS), lifestyle changes, and treatment adherence for use in Brazil. The questionnaire has three scales: general knowledge of risk factors (RFs); specific knowledge of these RFs; and lifestyle changes achieved. Cross-cultural adaptation included translation, synthesis, back-translation, expert committee review, and pretesting. Face and content validity, reliability, and construct validity were measured. Cronbach’s alpha for the total sample (n = 240) was 0.75. Assessment of psychometric properties revealed adequate face and content validity, and the construct revealed seven components. It was concluded that the Brazilian version of Q-FARCS had adequate reliability and validity for the assessment of knowledge of cardiovascular RFs.

Disponibilidade de nitrogênio e fluxos de N2O a partir de solo sob pastagem após aplicação de herbicida

O óxido nitroso (N2O) é um importante gás do efeito estufa, com um alto potencial de provocar o aquecimento global e de vida longa na atmosfera. Grande parte dos fluxos naturais de N2O ocorre a partir dos oceanos, enquanto o restante é resultado principalmente da contribuição de processos microbiológicos (nitrificação e desnitrificação) ocorridos em solos de regiões tropicais. A disponibilidade de N para processos metabólicos dos microrganismos pode ser um importante fator no controle das emissões de N2O nesses solos. O presente trabalho foi realizado com o objetivo de melhorar a compreensão acerca dos controles sobre os fluxos de N2O a partir de solos com pastagem na Amazônia. Foram investigados os efeitos da disponibilidade de N, C e umidade na emissão de N2O a partir do solo de uma pastagem submetida ou não a práticas para recuperação de sua produtividade. O estudo foi realizado em condições de laboratório, incubando o solo coletado nas seguintes situações: (a) Controle - solo com pastagem estabelecida em 1983 e com manejo tradicional, constituída de uma mistura de Brachiaria brinzantha e Panicum maximum, além da presença de uma variedade de plantas invasoras, e (b) Herbicida - dessecação total das plantas da pastagem estabelecida em 1983, para o plantio direto de arroz e posterior ressemeadura de braquiária. É oportuno ressaltar que as amostras de solo (camada de 0-5 cm) das áreas foram coletadas no terceiro dia após a aplicação do agroquímico. Para o estabelecimento das comparações e compreensão dos controles nos fluxos de N2O do solo, foram adicionados ao solo para incubação: nitrato, dextrose e água; além do uso de acetileno para bloquear a oxidação do N2O a N2, estimando, assim, a proporção de N2 que foi emitido do solo. Os maiores fluxos foram observados quando o nitrato foi acrescentado ao solo sob condições de alta umidade. A adição de dextrose (fonte de C) elevou os fluxos de forma mais intensa no solo que recebeu aplicação de herbicida, cuja disponibilidade de N também era maior. Com a aplicação de acetileno foi possível observar que grande parte do N perdido sob formas gasosas ocorre como N2. Desse modo, o processo de desnitrificação mostrou-se dominante nos fluxos de N2O a partir do solo da pastagem estudada, enquanto o N foi o fator principal no controle desses fluxos.

Qualidade da matéria orgânica e estoques de carbono e nitrogênio em Latossolo tratado com biossólido e cultivado com eucalipto

O objetivo deste estudo foi determinar o efeito da aplicação de doses de um biossólido alcalino nos estoques de C e N, bem como na qualidade da matéria orgânica (MO) de um Latossolo Vermelho-Amarelo distrófico cultivado com eucalipto, após cinco anos da aplicação do resíduo. O trabalho foi desenvolvido em área pertencente à Estação Experimental da ESALQ/USP, no município de Itatinga, Estado de São Paulo. Em março de 1998, após corte de uma plantação de Eucalyptus grandis com sete anos de idade e novo plantio, com a mesma espécie, no sistema de cultivo mínimo, foi iniciado o experimento. Quatro meses depois, foi aplicado o biossólido digerido anaeróbio, com umidade original, sobre a superfície do solo, nas entrelinhas da cultura e sem posterior incorporação. Cinco tratamentos foram avaliados: (a) controle; (b) fertilização mineral com N, P, K, B e Zn (Fert. Mineral); (c) 10 t ha-1 de biossólido + K (10 t ha-1 + K); (d) 20 t ha-1 de biossólido + K (20 t ha-1 + K); e (e) 40 t ha-1 de biossólido + K (40 t ha-1 + K). Amostras de solo das camadas de 0-5, 5-10, 10-20, 20-30 e 30-60 cm foram coletadas em setembro de 2003, cerca de cinco anos após a aplicação do resíduo. Para todas as profundidades foram determinados os teores totais de C e N e a densidade do solo. Nas amostras coletadas até 20 cm de profundidade, foram feitas também determinações de pH, frações de carbono orgânico (CO) por graus de oxidação, teores de açúcares solúveis, proteína bruta, lipídeos, hemicelulose, celulose e lignina, CTC potencial (a pH 7,0) e CTC efetiva (ao pH natural). Após cinco anos da aplicação do biossólido, não foram observadas diferenças entre os tratamentos, para os teores totais e estoques de C e N, densidade e frações do CO. Os estoques médios de C foram iguais a 26,52; 7,96; 10,31; 12,88; e 31,19 t ha-1; e de N iguais a 0,97; 0,39; 0,99; 0,63; e 0,34 t ha-1, respectivamente, para as camadas de 0-5, 5-10, 10-20, 20-30 e 30-60 cm. Cerca de 50 % do total de CO esteve presente no compartimento denominado lábil, comportamento típico de áreas com espécies e, ou, manejo que favorecem o retorno de resíduos vegetais ao solo. Dos compostos orgânicos determinados, somente a lignina mostrou alteração de acordo com os tratamentos. Os tratamentos 40 t ha-1 + K e Fert. Mineral apresentaram a MO do solo na camada de 0-5 cm mais enriquecida com lignina, em comparação aos demais, sendo esse efeito atribuído à maior deposição de folhas nesses dois tratamentos e à natureza recalcitrante da lignina. Os resultados de CTC não evidenciaram efeitos dos tratamentos na qualidade da MO, pelo menos no que se refere a esta propriedade. A CTC (pH natural) mostrou-se mais dependente dos valores de pH do solo do que dos teores de C.