Morfohistología testicular de Ceratophrys ornata (Bell) (Amphibia, Anura, Ceratophryidae)

En el presente trabajo se describen las características morfohistológicas del testículo de individuos adultos (n=5) de Ceratophrys ornata (Bell, 1843) provenientes de humedales del centro de Argentina. Los mismos se procesaron mediante técnicas histológicas de rutina, se cortaron a 8 µm y las láminas obtenidas se tiñeron con hematoxilina-eosina y tricrómico de Masson. Las gónadas son órganos pares, amarillentos, alargados y contorneados de 18,58 ± 0,23 mm de largo por 1,51 ± 0,13 mm de ancho. Histológicamente se observa una delgada túnica albugínea (6,29 ± 0,83 µm) rodeando a los testículos. En su interior se hallan lóculos seminíferos que miden 240,64 ± 38,52 µm de diámetro, en ellos se distinguen cistos con células espermatogénicas en distintas etapas de desarrollo. El tejido intersticial es escaso y en él se destacan las células de Leydig y vasos sanguíneos. Las espermatogonias I son las células más grandes de la serie germinal (20,03 ± 2,27 µm); poseen la cromatina granular y de aspecto multilobular, hallándose comúnmente una por cisto, estas originan a las espermatogonias II, más pequeñas (12,06 ± 1,14 µm). Los espermatocitos I presentan la cromatina levemente condensada y son un poco más chicos que sus precedentes (11,64 ± 0,36 µm). Los espermatocitos II miden 8,85 ± 0,54 µm. Las espermátidas I son esféricas, miden 5,95 ± 0,42 µm y se agrupan en cistos redondeados. Las espermátidas II, en cambio son alargadas y no se hallan dentro de cistos, pero siguen organizadas en paquetes asociadas a células de Sertoli. Los espermatozoides son células libres hacia el centro del lóculo, alargadas, flageladas y con una notable compactación nuclear. La morfohistología de los testículos analizados muestran características macroscópicas e histológicas similares a las observadas en otras especies de anfibios anuros neotropicales, presentando todas las células del linaje espermatogénico en un mismo lóculo, lo que indicaría que presentan ciclos espermatogénicos continuos.

Trypanosomatidae codon usage and GC distribution

A study od Typanosomatidae GC distribution and codon usage is presented. The codon usage patterns in coincidence with the phylogenetical data are similar in Crithidia and Leishmania, whereas they are more divergent in Trypanosoma brucei and T. cruzi. The analyisis of the GC mutational pressure in these organisms reveals that T. brucei, and to a lesser extent T. cruzi, have envolved towards a more balanced use of all bases, whereas Leishmania and Crithidia retain features of a primeval genetic apparatus. Tables with approximated GC mutational pressure in homologous genes, and codon usage in Trypanosomatidae are presented.

Towards the Physical Map of the Trypanosoma cruzi Nuclear Genome: Construction of YAC and BAC Libraries of the Reference Clone T. cruzi CL-Brener

Strategies to construct the physical map of the Trypanosoma cruzi nuclear genome have to capitalize on three main advantages of the parasite genome, namely (a) its small size, (b) the fact that all chromosomes can be defined, and many of them can be isolated by pulse field gel electrophoresis, and (c) the fact that simple Southern blots of electrophoretic karyotypes can be used to map sequence tagged sites and expressed sequence tags to chromosomal bands. A major drawback to cope with is the complexity of T. cruzi genetics, that hinders the construction of a comprehensive genetic map. As a first step towards physical mapping, we report the construction and partial characterization of a T. cruzi CL-Brener genomic library in yeast artificial chromosomes (YACs) that consists of 2,770 individual YACs with a mean insert size of 365 kb encompassing around 10 genomic equivalents. Two libraries in bacterial artificial chromosomes (BACs) have been constructed, BACI and BACII. Both libraries represent about three genome equivalents. A third BAC library (BAC III) is being constructed. YACs and BACs are invaluable tools for physical mapping. More generally, they have to be considered as a common resource for research in Chagas disease

Genes and Chromosomes of Leishmania infantum

During recent years, several Leishmania infantum genes have been cloned and characterized. Here, we have summarized the available information on the gene organization and expression in this protozoan parasite. From a comparative analysis, the following outstanding features were found to be common to most of the genes characterized: tandemly organized genes with conserved coding regions and divergent untranslated regions, polycistronic transcription and post-transcriptional regulation of gene expression. The analysis of chromosomes of L. infantum by pulsed-field electrophoresis showed the existence of both size and number polymorphisms such that each strain has a distinctive molecular karyotype. Despite this variability, highly conserved physical linkage groups exists among different strains of L. infantum and even among Old World Leishmania species. Gene mapping on the L. infantum molecular karyotype evidenced a bias in chromosomal distribution of, at least, the evolutionary conserved genes

Molecular and Antigenic Characterization of the Leishmania (Viannia) panamensis Kinetoplastid Membrane Protein-11

The kinetoplastid membrane protein 11 (KMP-11) has been recently described in Leishmania (Leishmania) donovani as a major component of the promastigote membrane. Two oligonucleotide primers were synthesized to PCR-amplify the entire coding region of New World Leishmania species. The Leishmania (Viannia) panamensis amplification product was cloned, sequenced and the putative amino acid sequence determined. A remarkably high degree of sequence homology was observed with the corresponding molecule of L. (L) donovani and L. (L) infantum (97% and 96%, respectively). Southern blot analysis showed that the KMP-11 locus is conformed by three copies of the gene. The L. (V) panamensis ORF was subsequently cloned in a high expression vector and the recombinant protein was induced and purified from Escherichia coli cultures. Immunoblot analysis showed that 80%, 77% and 100% sera from cutaneous, mucocutaneous and visceral leishmaniasis patients, respectively, recognized the recombinant KMP-11 protein. In a similar assay, 86% of asymptomatic Leishmania-infected individuals showed IgG antibodies against the rKMP-11. We propose that KMP-11 could be used as a serologic marker for infection and disease caused by Leishmania in America.

Are vectors able to learn about their hosts? A case study with Aedes aegypti mosquitoes

The way in which vectors distribute themselves amongst their hosts has important epidemiological consequences. While the role played by active host choice is largely unquestioned, current knowledge relates mostly to the innate response of vectors towards stimuli signalling the presence or quality of their hosts. Many of those cues, however, can be unpredictable, and therefore prevent the incorporation of the appropriate response into the vector's behavioural repertoire unless some sort of associative learning is possible. We performed a wide range of laboratory experiments to test the learning abilities of the mosquito, Aedes aegypti. Mosquitoes were exposed to choice procedures in (1) an olfactomenter and (2) a 'visual arena'. Our goal was to determine whether the mosquitoes were able to associate unconditional stimuli (blood feeding, human breath, vibration and electrical shock) with particular odours (citral, carvone, citronella oil and eugenol) and visual patterns (horizontal or vertical black bars) to which they had been previously observed to be responsive. We found no evidence supporting the hypothesis that associative learning abilities are present in adult Ae. aegypti. We discuss the possibilities that the assays employed were either inappropriate or insufficient to detect associative learning, or that associative learning is not possible in this species.

Asymptomatic Plasmodium spp. infection in Tierralta, Colombia

With the aim of determining the prevalence of asymptomatic Plasmodium spp. infection by thick smear and PCR and its association with demographic and epidemiological characteristics in the village of Nuevo Tay, Tierralta, Córdoba, Colombia, a cross-sectional population study was carried out, using random probabilistic sampling. Venous blood samples were taken from 212 people on day 0 for thick smear and PCR. Clinical follow-up and thick smears were carried out on days 14 and 28. The prevalence of Plasmodium spp. infection was 17.9% (38/212; 95% CI: 12.5-23.3%) and the prevalence of asymptomatic Plasmodiumspp. infection was 14.6% (31/212; 95% CI: 9.6-19.6%). Plasmodium vivax was found more frequently (20/31; 64.5%) than Plasmodium falciparum (9/31; 29%) and mixed infections (2/31; 6.5%). A significantly higher prevalence of asymptomatic infection was found in men (19.30%) than in women (9.18%) (prevalence ratio: 2.10; 95% CI: 1.01-4.34%; p = 0.02). People who developed symptoms had a significantly higher parasitemia on day 0 than those who remained asymptomatic, of 1,881.5 ± 3,759 versus 79 ± 106.9 (p = 0.008). PCR detected 50% more infections than the thick smears. The presence of asymptomatic Plasmodium spp. infection highlights the importance of carrying out active searches amongst asymptomatic populations residing in endemic areas.

An evaluation of the dot-ELISA procedure as a diagnostic test in an area with a high prevalence of human Toxocara canis infection

The aim of this work was to evaluate a dot-enzyme-linked immunosorbent assay (dot-ELISA) using excretory-secretory antigens from the larval stages of Toxocara canis for the diagnosis of toxocariasis. A secondary aim was to establish the optimal conditions for its use in an area with a high prevalence of human T. canis infection. The dot-ELISA test was standardised using different concentrations of the antigen fixed on nitrocellulose paper strips and increasing dilutions of the serum and conjugate. Both the dot-ELISA and standard ELISA methods were tested in parallel with the same batch of sera from controls and from individuals living in the problem area. The best results were obtained with 1.33 µg/mL of antigen, dilutions of 1/80 for the samples and controls and a dilution of 1/5,000 for the anti-human IgG-peroxidase conjugate. All steps of the procedure were performed at room temperature. The coincidence between ELISA and dot-ELISA was 85% and the kappa index was 0.72. The dot-ELISA test described here is rapid, easy to perform and does not require expensive equipment. Thus, this test is suitable for the serological diagnosis of human T. canis infection in field surveys and in the primary health care centres of endemic regions.

Identification of blood meal sources of Lutzomyia longipalpis using polymerase chain reaction-restriction fragment length polymorphism analysis of the cytochrome B gene

An analysis of the dietary content of haematophagous insects can provide important information about the transmission networks of certain zoonoses. The present study evaluated the potential of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis of the mitochondrial cytochrome B (cytb) gene to differentiate between vertebrate species that were identified as possible sources of sandfly meals. The complete cytb gene sequences of 11 vertebrate species available in the National Center for Biotechnology Information database were digested with Aci I, Alu I, Hae III and Rsa I restriction enzymes in silico using Restriction Mapper software. The cytb gene fragment (358 bp) was amplified from tissue samples of vertebrate species and the dietary contents of sandflies and digested with restriction enzymes. Vertebrate species presented a restriction fragment profile that differed from that of other species, with the exception of Canis familiaris and Cerdocyon thous. The 358 bp fragment was identified in 76 sandflies. Of these, 10 were evaluated using the restriction enzymes and the food sources were predicted for four: Homo sapiens (1), Bos taurus (1) and Equus caballus (2). Thus, the PCR-RFLP technique could be a potential method for identifying the food sources of arthropods. However, some points must be clarified regarding the applicability of the method, such as the extent of DNA degradation through intestinal digestion, the potential for multiple sources of blood meals and the need for greater knowledge regarding intraspecific variations in mtDNA.

Getting ready for malaria elimination: a check list of critical issues to consider

In recent years, a renewed interest in malaria elimination and eradication has emerged and seems to be rooting in the minds of the scientific community, public health specialists, funding bodies, policy makers and politicians. Malaria eradication will certainly benefit from improved and innovative tools; notwithstanding novel knowledge in fields ranging from basic science to mathematical modelling and health systems research. However, the elimination of malaria also encompasses a broad range of essential aspects that countries and other actors need to consider when thinking of embarking on such an adventure, including the implementation of innovative strategies, the ability to incorporate the most up-to-date evidence into policy, the integration of malaria into the broader health agenda, the strengthening of surveillance and health systems, capacity building, funding, advocacy and, very importantly, research. While in some cases this enthusiasm is clearly justified, some countries are still a long way from realistically advancing towards elimination. This paper attempts to provide guidance on all the necessary issues that should be considered when initiating a malaria elimination program.

Construction of three new Gateway® expression plasmids for Trypanosoma cruzi

We present here three expression plasmids for Trypanosoma cruzi adapted to the Gateway® recombination cloning system. Two of these plasmids were designed to express trypanosomal proteins fused to a double tag for tandem affinity purification (TAPtag). The TAPtag and Gateway® cassette were introduced into an episomal (pTEX) and an integrative (pTREX) plasmid. Both plasmids were assayed by introducing green fluorescent protein (GFP) by recombination and the integrity of the double-tagged protein was determined by western blotting and immunofluorescence microscopy. The third Gateway adapted vector assayed was the inducible pTcINDEX. When tested with GFP, pTcINDEX-GW showed a good response to tetracycline, being less leaky than its precursor (pTcINDEX).

Explicações para a divulgação voluntária ambiental no Brasil utilizando a análise de regressão em painel

O objetivo no estudo aqui apresentado foi identificar os fatores que determinam a divulgação voluntária ambiental pelas empresas brasileiras potencialmente poluidoras. Para tanto, foram analisa- das as Demonstrações Financeiras Padronizadas (DFPs) e os Relató- rios de Sustentabilidade (RS) do período de 2005 a 2007 das empresas abertas com ações listadas na Bolsa de Valores de São Paulo (Bovespa) e pertencentes a setores de alto impacto ambiental, que compreendem extração e tratamento de minerais; metalúrgico; químico; papel e celulose; indústria de couros e peles; transporte, terminais, depósitos e comércio (de combustíveis, derivados de petróleo e produtos químicos). Com o intuito de explicar a evidenciação ambiental divulgada pelas empresas investigadas, foram formuladas sete hipóteses testadas a partir de instrumentos de análise estatística. Essas hipóteses referem-se a fatores individuais das empresas, que englobam tamanho, rentabilidade, endividamento, empresa de auditoria, sustentabilidade, internacionalização e publicação do RS. Os resultados mostram que, nos três anos analisados, as 57 empresas que compõem a amostra do estudo evidenciaram um total de 6.182 sentenças ambientais, 73% delas divulgadas nos RS e 27% nas DFPs. A análise de regressão em painel demonstrou que as variáveis tamanho da empresa, empresa de auditoria, sustentabilidade e publicação do RS são relevantes a um nível de significância de 5% para a explicação do disclosure voluntário de informações ambientais. Concluiu-se que os achados da pesquisa corroboram a teoria positiva da contabilidade, e parcialmente a teoria da legitimidade.

Atenção integral na saúde do idoso no Programa Saúde da Família: visão dos profissionais de saúde

O Brasil, até o ano de 2025, será o sexto país do mundo em número de idosos. É função das políticas públicas contribuir para que as pessoas alcancem idades avançadas com melhor saúde. O objetivo deste estudo foi conhecer o que pensam os profissionais sobre necessidades de saúde e atenção integral. Os sujeitos foram dezessete, que compõem duas equipes do Programa Saúde da Família, no Município de Santos. A abordagem foi qualitativa, os discursos foram analisados a partir da Representação Social, referencial teórico do Discurso do Sujeito Coletivo. A análise revelou que os profissionais se esforçam para atender as necessidades de saúde dos idosos através de ações de prevenção e promoção da saúde. No entanto, suas necessidades sociais e psicológicas não estão sendo atendidas. Concluímos que os idosos necessitam de maior integralidade e agilidade no sistema de saúde, porque possuem dificuldades no acesso e deslocamento nos serviços de saúde.

Actitud ante el consumo y no consumo de alcohol en estudiantes de preparatoria – México

El objetivo del estudio fue identificar diferencias de actitud entre estudiantes de la educación secundaria consumidores y no de alcohol a través de la Teoría de la Conducta Planeada. Estudio descriptivo transversal con abordaje cuantitativo, con una muestra de 131 adolescentes. Se verifi có que 74% ya habían consumido alcohol y 18.3% tienen consumo dañino. Se constató que las creencias conductuales hacia el consumo eran más altas en los consumidores de alcohol (X=29.32, Mdn=27.50) que en los no consumidores, así con la evaluación de las creencias en sentido positivo son más altas en los consumidores de alcohol (X=17.72, Mdn=9.52) que en los no consumidores, haciéndose necesaria la implementación de programas preventivos que fortalezcan las creencias de los adolescentes, así como estimular las estrategias vinculadas con el fortalecimiento de factores protectores y estilos de vida saludables.

Fruit flies (Diptera, Tephritidae) and their parasitoids on cultivated and wild hosts in the Cerrado-Pantanal ecotone in Mato Grosso do Sul, Brazil

Fruit flies (Diptera, Tephritidae) and their parasitoids on cultivated and wild hosts in the Cerrado-Pantanal ecotone in Mato Grosso do Sul, Brazil. Information on frugivorous flies in cultivated or wild host plants and their parasitoids in the Cerrado-Pantanal ecotone in Aquidauana, Mato Grosso do Sul is presented and discussed. Fruit fly samples were collected weekly in specific fruit trees, and McPhail® traps were installed in the same trees for a period of two years. The fruit flies infested ripe and unripe fruits of Averrhoa carambola L., Schoepfia sp., Psidium guajava L. and Pouteria torta (Mart.) Radlk and mature fruits of Anacardium occidentale L. and Inga laurina (Sw.) Willd. Nineteen fruit fly species were obtained with the combination of sampling methods (collecting fruits and trapping), nine of them obtained with both methods, five found only in fruits and five only in traps. This is the first record of Anastrepha striata Schiner in a species of Sapotaceae, as well as for A. castanea Norrbom and A. daciformes Bezzi in Schoepfia sp. (Olacaceae), and for A. distincta Greene in fruits of P. guajava in the state of Mato Grosso do Sul. Fruit collections simultaneously associated with capture of fruit flies by McPhail traps in the same host plants are essential to understand the diversity of fruit flies and their relationship with hosts and parasitoids. Species of Braconidae and Pteromalidae were recovered, where Doryctobracon areolatus (Szépligeti) was the most abundant parasitoid in larvae of tephritids infesting both cultivated and wild host fruits.