Characterization and Comparison of Genetic Variation in Cotesia flavipes (Hymenoptera: Braconidae) Mass Reared for Biological Pest Control Using Microsatellite Markers.

The larval parasitoid Cotesia flavipes (Cameron) (Hymenoptera: Braconidae) is an important biological control agent of the sugarcane borer, Diatraea saccharalis (Fabricius) (Lepidoptera: Crambidae). This exotic parasitoid has been mass reared for field release since its introduction in the 1970s. Insects are exchanged between labs or introduced from the field, but without basic studies or criteria. Thus, the objective of this study was to evaluate the genetic variability of insects from six Brazilian States (São Paulo, Minas Gerais, Paraná, Goiás, Maranhão, and Alagoas) using microsatellites. Analysis of five loci using at least 22 females from each location was performed. The molecular analysis made possible to verify that four out of the five loci analyzed were polymorphic. The allele frequencies of three loci were in agreement with the Hardy-Weinberg Equilibrium for all insects of all regions. It was also observed that five alleles were exclusively presented in only two loci. The variation among and within populations was 24.65 and 75.34%, respectively. The estimated shared genotypes between the C. flavipes individuals showed that K = 2 was the most likely number of genetic groups causing the current variation, as well as high shared genotypes from these groups of the individuals. Considering all the analyzed loci, the genetic differentiation was at a moderate level. We suggest a possible mixture of biological materials based on genetic distances and the degree of structuring displayed.

Efficacy of Soybean's Event DAS-81419-2 Expressing Cry1F and Cry1Ac to Manage Key Tropical Lepidopteran Pests Under Field Conditions in Brazil.

Bacillus thuringiensis (Bt) event DAS-81419-2 (Conkesta technology) in soybean, Glycine max (L.) Merrill, expresses Cry1F and Cry1Ac proteins to provide protection from feeding by several lepidopteran pests. A total of 27 field experiments across nine locations were conducted from 2011 to 2015 in southern and central Brazil to characterize the efficacy of DAS-81419-2 soybean infested with Anticarsia gemmatalis Hübner (Lepidoptera: Erebidae), Chrysodeixis includens (Walker) (Lepidoptera: Noctuidae), Heliothis virescens (F.) (Lepidoptera: Noctuidae), and Spodoptera cosmioides (Walker) (Lepidoptera: Noctuidae) during vegetative (V4) and reproductive (R2 and R4) crop developmental stages. The efficacy of DAS-81419-2 was compared to that of a non-Bt isogenic variety managed with or without applications of commercial foliar insecticides for lepidopteran control. DAS-81419-2 soybean consistently experienced defoliation levels of 0.5% or less (compared with 20.05-56.74% in the non-Bt, nonsprayed treatment) and larval survival of < 0.1% in all four species across the vegetative and reproductive plant stages evaluated. The efficacy of DAS-81419-2 was significantly higher than commercial foliar insecticides applied to the non-Bt variety. DAS-81419-2 soybeans containing two highly effective Bt proteins are expected to be a more robust IRM tool compared to single-trait Bt technologies. The consistent efficacy of DAS-81419-2 soybeans across years, locations, and crop stages suggests that it will be a valuable product for management of hard-to-control key lepidopteran pests in South American soybean production.

Fisetin as a promising antifungal agent against Cryptocococcus neoformans species complex.

AIMS: The aim of this study was to investigate the mechanisms of action of fisetin, a flavonol with antifungal activity previously evaluated against the Cryptococcus neoformans species complex. METHODS AND RESULTS: Ergosterol content and flow cytometry analysis were determined for the C. neoformans species complex in the presence of fisetin and ultrastructural analysis of morphology was performed on Cryptococcus gattii and C. neoformans. Decrease in the total cellular ergosterol content after exposure to fisetin ranged from 25·4% after exposure to 128 µg ml(-1) to 21·6% after exposure to 64 µg ml(-1) of fisetin compared with the control (without fisetin). The fisetin effects obtained with flow cytometry showed metabolic impairment, and alterations in its normal morphology caused by fisetin in C. neoformans cells were verified using scanning electron microscopy. CONCLUSIONS: Fisetin is a compound that acts in the biosynthesis of ergosterol. Flow cytometry showed that fisetin reduced viability of the metabolically active cells of C. gattii, while morphological changes explain the action of fisetin in inhibiting growth of these fungi. SIGNIFICANCE AND IMPACT OF THE STUDY: This study supports the idea that fisetin may represent a good starting point for the development of future therapeutic substances for cryptococcosis.

Molecular and phylogenetic characterization based on the complete genome of a virulent pathotype of Newcastle disease virus isolated in the 1970s in Brazil.

Newcastle disease (ND) is caused by the avian paramyxovirus type 1 (APMV-1) or Newcastle disease virus (NDV) that comprises a diverse group of viruses with a single-stranded, negative-sense RNA genome. ND is one of the most important diseases of chickens, because it severely affects poultry production worldwide. In the 1970s, outbreaks of virulent ND were recorded in Brazil, and the strain APMV-1/Chicken/Brazil/SJM/75 (SJM) of NDV was isolated. This strain was characterized as highly pathogenic for chickens but not pathogenic for other bird species. Here we present the complete genome of NDV strain SJM and investigate the phylogenetic relationships of this virus with other NDV strains in terms of genome and proteins composition, as well as characterizing its evolution process. The NDV strain SJM is categorized as a velogenic virus and the complete genome is 15,192 nucleotides in length, consisting of six genes in the order 3'-NP-P-M-F-HN-L-5'. The presence of the major pathogenic determinant of NDV strains ((112)R-R-Q-K-R↓F(117)) was identified in the Fusion protein of the NDV strain SJM. In addition, phylogenetic analysis classified the NDV strain SJM as a member of class II, genotype V, and indicates that this virus help us in the understanding of the evolutionary process of strains belonging to this genotype. This study contributes to the growing interest involving the characterization of NDV isolates to improve our current understanding about the epidemiology, surveillance and evolution of the pathogenic strains.

Alcohol intake in lactating women assisted in a University Hospital.

OBJECTIVE: To determine the prevalence of alcohol intake and the degree of alcohol-related risk among nursing mothers attended at the Child Care Service of Hospital das Clínicas of Universidade Federal de Pernambuco, Brazil. METHODS: A cross-sectional study was carried out with 157 nursing mothers enrolled in the Child Care Program of the university hospital. A questionnaire was administered addressing demographic and socioeconomic variables, type and duration of breastfeeding, smoking habits and consumption of foods considered as appetizers. The Alcohol Use Disorders Identification Test (AUDIT C) was applied for assessing alcohol consumption in the previous 12 months. Pearson's chi-square test and Fisher's exact test were used for statistical analysis. RESULTS: Twelve percent of the nursing mothers reported consuming alcoholic beverages, 100% of whom were classified as being at low risk for alcohol use disorders. The frequency of nursing mothers who consumed appetizers during alcohol consumption was 100%, the most common of which was cheese - 18 (95%). CONCLUSIONS: The prevalence of alcohol intake was low in the nursing mothers analyzed. The users exhibited a low risk for alcohol disorders and a high frequency of the consumption of appetizers during alcohol consumption.

Ingestão de bebidas alcoólicas em lactantes atendidas em Hospital Universitário / Alcohol intake in lactating women assisted in a University Hospital

OBJETIVOS: Verificar a frequência do consumo de bebidas alcoólicas e o grau de risco do hábito de etilismo em lactantes atendidas no Serviço de Puericultura do Hospital das Clínicas da Universidade Federal de Pernambuco. MÉTODOS: Estudo transversal com 157 mães lactantes matriculadas no Programa de Puericultura do hospital. Utilizou-se questionário com informações sobre variáveis demográficas, socioeconômicas, tipo e tempo de aleitamento materno, hábito de tabagismo e consumo de alimentos considerados petiscos. A avaliação do consumo de álcool nos últimos 12 meses foi realizada pelo questionário "teste de identificação de distúrbios causados pelo uso de álcool" (AUDIT C - Alcohol Use Disorders Identification Test). Para análise estatística, aplicaram-se os testes do qui-quadrado de Pearson e exato de Fisher. RESULTADOS: Verificou-se consumo de bebidas alcoólicas em 12% das lactantes; dentre estas, 100% apresentaram baixo risco para transtornos causados pelo uso do álcool. A frequência de lactantes que consumiam petiscos durante a ingestão alcoólica foi 100%, sendo os queijos os mais consumidos - 18 (95%). CONCLUSÕES: A prevalência de consumo de álcool foi baixa nas lactantes estudadas. As usuárias exibiram um consumo considerado de baixo risco e uma frequência elevada de consumo de petiscos durante a ingestão alcoólica.

OBJECTIVE: To determine the prevalence of alcohol intake and the degree of alcohol-related risk among nursing mothers attended at the Child Care Service of Hospital das Clínicas of Universidade Federal de Pernambuco, Brazil. METHODS: A cross-sectional study was carried out with 157 nursing mothers enrolled in the Child Care Program of the university hospital. A questionnaire was administered addressing demographic and socioeconomic variables, type and duration of breastfeeding, smoking habits and consumption of foods considered as appetizers. The Alcohol Use Disorders Identification Test (AUDIT C) was applied for assessing alcohol consumption in the previous 12 months. Pearson's chi-square test and Fisher's exact test were used for statistical analysis. RESULTS: Twelve percent of the nursing mothers reported consuming alcoholic beverages, 100% of whom were classified as being at low risk for alcohol use disorders. The frequency of nursing mothers who consumed appetizers during alcohol consumption was 100%, the most common of which was cheese - 18 (95%). CONCLUSIONS: The prevalence of alcohol intake was low in the nursing mothers analyzed. The users exhibited a low risk for alcohol disorders and a high frequency of the consumption of appetizers during alcohol consumption.

OBJETIVOS: Verificar la frecuencia del consumo de bebidas alcohólicas y el grado de riesgo del hábito de alcoholismo en lactantes atendidas en el Servicio de Cuidado del Niño del Hospital de Clínicas de la Universidad Federal de Pernambuco (HC/UFPE). MÉTODO: Estudio transversal con 157 madres lactantes matriculadas en el Programa de Cuidado del Niño del HC. Se utilizó cuestionario con informaciones sobre variables demográficas, socioeconómicas, tipo y tiempo de lactancia materna, hábito de tabaquismo y consumo de alimentos considerados bocados. La evaluación del consumo de alcohol en los últimos 12 meses fue realizada por el cuestionario AUDIT C (alcohol use disorders identification test). Para el análisis estadístico se utilizaron las pruebas de Chi-cuadrado de Pearson y Exacto de Fisher. RESULTADOS: El consumo de bebidas alcohólicas fue verificado en 12% de las lactantes, siendo en 100% de ellas clasificado de bajo riesgo para trastornos causados por el uso del alcohol. La frecuencia de lactantes que consumían bocados durante la ingestión alcohólica fue del 100%, siendo los quesos los más consumidos 18 (94,7%). CONCLUSIONES: La prevalencia de consumo de alcohol fue baja en las lactantes estudiadas. Las usuarias mostraron un consumo considerado de bajo riesgo y una frecuencia elevada de consumo de bocados durante la ingestión alcohólica.

Bacillus thuringiensis Cry1Ia10 and Vip3Aa protein interactions and their toxicity in Spodoptera spp. (Lepidoptera).

The polyphagous pests belonging to the genus Spodoptera are considered to be among the most important causes of damage and are widely distributed throughout the Americas'. Due to the extensive use of genetically modified plants containing Bacillus thuringiensis genes that code for insecticidal proteins, resistant insects may arise. To prevent the development of resistance, pyramided plants, which express multiple insecticidal proteins that act through distinct mode of actions, can be used. This study analyzed the mechanisms of action for the proteins Cry1Ia10 and Vip3Aa on neonatal Spodoptera frugiperda, Spodoptera albula, Spodoptera eridania and Spodoptera cosmioides larvae. The interactions of these toxins with receptors on the intestinal epithelial membrane were also analyzed by binding biotinylated toxins to brush border membrane vesicles (BBMVs) from the intestines of these insects. A putative receptor of approximately 65 kDa was found by ligand blotting in all of these species. In vitro competition assays using biotinylated proteins have indicated that Vip3Aa and Cry1Ia10 do not compete for the same receptor for S. frugiperda, S. albula and S. cosmioides and that Vip3Aa was more efficient than Cry1Ia10 when tested individually, by bioassays. A synergistic effect of the toxins in S. frugiperda, S. albula and S. cosmioides was observed when they were combined. However, in S. eridania, Cry1Ia10 and Vip3Aa might compete for the same receptor and through bioassays Cry1Ia10 was more efficient than Vip3Aa and showed an antagonistic effect when the proteins were combined. These results suggest that using these genes to develop pyramided plants may not prove effective in preventing the development of resistance in S. eridiana.

Antifungal susceptibility patterns of yeasts and filamentous fungi isolated from nail infection.

BACKGROUND: Onychomycosis is the nail infection caused by a wide spectrum of fungi species, including yeasts, dermatophytes and filamentous fungi non-dermatophytes (FFND). This fungal infection represents an important medical problem because it involves the patient's life quality. OBJECTIVE: The aim was to isolate and identify the fungal agents of onychomycosis, and to determine the in vitro susceptibility to antifungal agents. METHODS: During the period of March 2008 to March 2009, 114 patients clinically suspected of having onychomycosis were examined. Demographic data, mainly age and gender were obtained from each patient. The nail samples collected (136) were submitted to direct examination with potassium hydroxide 20% and grown on Sabouraud dextrose agar. The in vitro antifungal susceptibility testing was performed according to the method of broth microdilution, recommended by the Clinical Laboratory Standards Institute (CLSI). RESULTS: Onychomycosis was observed in 95 (83.3%) patients, including 16 men (16.8%) and 79 women (83.2%), with mean age of 48.1 years. Candida parapsilosis, Trichophyton rubrum and Fusarium spp were the fungi most frequently isolated. The most of the isolated yeasts showed susceptibility to antifungal agents studied. Among filamentous fungi, high MIC values to itraconazole were found for T. rubrum and T. mentagrophytes, while Fusarium spp showed decreased susceptibility to itraconazole and voriconazole. CONCLUSION: C. parapsilosis was the most common fungal species isolated from patients with onychomycosis. The different response obtained by in vitro susceptibility testing to drugs shows the importance of these methods to assist clinicians in choosing the best therapeutic option.

Interação interespecífica entre o parasitoide Telenomus remus Nixon (Hymenoptera: Scelionidae) e o predador Doru luteipes Scudder (Dermaptera: Forficulidae) sobre ovos de Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae) / Interaction between Telenomus remus Nixon (Hymenoptera: Scelionidae) and Doru luteipes Scudder (Dermaptera: Forficulidae) on Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae) eggs

Este trabalho teve por objetivo verificar a interação entre o parasitoide Telenomus remus e o predador Doru luteipes. Para tanto, posturas deS. frugiperda parasitadas porT. remus foram ofertadas a D. luteipes e ovos de D. luteipes foram ofertados a T. remus. Essa oferta levou em consideração diferentes fases do desenvolvimento embrionário do parasitoide e do predador. Constatou-se que quando as posturas de S. frugiperda foram expostas primariamente a T. remus. D. luteipes as consumiu, mas somente até o terceiro dia após o parasitismo.T. remus parasitou ovos deD. luteipes com até 48h de desenvolvimento embrionário, mas apenas quando as fêmeas do predador estavam ausentes. Diante disso, a interação intraguilda parece não ser um fator importante de regulação populacional destas espécies.

This work aimed at evaluating the interaction between Telenomus remus andD. luteipes. S. frugiperda eggs previously parasitized by T. remus were offered to D. luteipes and D. luteipes eggs were offered to T. remus. To study the influence of different embryonic development stages of the parasitoid, we offered S. frugiperda previously parasitized eggs at different embryonic development stages to the predator. And we offered predator eggs at different embryonic development stages to the parasitoid. The behavior of insects was observed. When S. frugiperda eggs were exposed to T. remus previously, D. luteipes eat the eggs until three days after parasitism. T. remus parasitized D. luteipes eggs until 48h they were laid, but only when predator females were absent.

Epidemiological and mycological data of onychomycosis in Goiania, Brazil.

Onychomycosis defined as fungal infection of the nail represents more than 50% of all onychopathies. Epidemiological studies have shown that this mycosis is worldwide in occurrence, but with geographical variation in distribution. The direct microscopy and culture of the nail samples were performed to identify the causative agent. Out of 2273 patients with nail infection examined between January 2000 and December 2004 in Goiania, state of Goias, Brazil, diagnosis of onychomycosis was confirmed in 1282 cases, with dermatophytes and Candida species being the most common aetiological agents isolated. Dermatophyte onychomycosis was more common in toenails than in fingernails, while onychomycosis caused by yeast had a similar frequency in both toenails and fingernails. Among the species identified, Candida albicans was responsible for 492 cases (38.4%) of onychomycosis, Trichophyton rubrum was found in 327 cases (25.6%) and Trichophyton mentagrophytes in 258 cases (20.1%). Other fungi isolated from nail infections included Aspergillus sp., Trichosporon sp., Geotrichum sp. and Fusarium sp. In our study, yeast of the genus Candida were the dominant cause of onychomycosis in women and dermatophytes were the principal cause of this condition in men.

INTERAÇÃO INTERESPECÍFICA ENTRE O PARASITOIDE TELENOMUS REMUS NIXON (HYMENOPTERA: SCELIONIDAE) E O PREDADOR DORU LUTEIPES SCUDDER (DERMAPTERA: FORFICULIDAE) SOBRE OVOS DE SPODOPTERA FRUGIPERDA (J.E. SMITH) (LEPIDOPTERA: NOCTUIDAE)

ABSTRACT This work aimed at evaluating the interaction between Telenomus remus andD. luteipes. S. frugiperda eggs previously parasitized by T. remus were offered to D. luteipes and D. luteipes eggs were offered to T. remus. To study the influence of different embryonic development stages of the parasitoid, we offered S. frugiperda previously parasitized eggs at different embryonic development stages to the predator. And we offered predator eggs at different embryonic development stages to the parasitoid. The behavior of insects was observed. When S. frugiperda eggs were exposed to T. remus previously, D. luteipes eat the eggs until three days after parasitism. T. remus parasitized D. luteipes eggs until 48h they were laid, but only when predator females were absent.

RESUMO Este trabalho teve por objetivo verificar a interação entre o parasitoide Telenomus remus e o predador Doru luteipes. Para tanto, posturas deS. frugiperda parasitadas porT. remus foram ofertadas a D. luteipes e ovos de D. luteipes foram ofertados a T. remus. Essa oferta levou em consideração diferentes fases do desenvolvimento embrionário do parasitoide e do predador. Constatou-se que quando as posturas de S. frugiperda foram expostas primariamente a T. remus. D. luteipes as consumiu, mas somente até o terceiro dia após o parasitismo.T. remus parasitou ovos deD. luteipes com até 48h de desenvolvimento embrionário, mas apenas quando as fêmeas do predador estavam ausentes. Diante disso, a interação intraguilda parece não ser um fator importante de regulação populacional destas espécies.

A new consensus for Trypanosoma cruzi intraspecific nomenclature: second revision meeting recommends TcI to TcVI.

In an effort to unify the nomenclature of Trypanosoma cruzi, the causative agent of Chagas disease, an updated system was agreed upon at the Second Satellite Meeting. A consensus was reached that T. cruzi strains should be referred to by six discrete typing units (T. cruzi I-VI). The goal of a unified nomenclature is to improve communication within the scientific community involved in T. cruzi research. The justification and implications will be presented in a subsequent detailed report.

A new consensus for Trypanosoma cruzi intraspecific nomenclature: second revision meeting recommends TcI to TcVI

In an effort to unify the nomenclature of Trypanosoma cruzi, the causative agent of Chagas disease, an updated system was agreed upon at the Second Satellite Meeting. A consensus was reached that T. cruzi strains should be referred to by six discrete typing units (T. cruzi I-VI). The goal of a unified nomenclature is to improve communication within the scientific community involved in T. cruzi research. The justification and implications will be presented in a subsequent detailed report.

Acute acquired toxoplasmosis: clinical-laboratorial aspects and ophthalmologic evaluation in a cohort of immunocompetent patients.

Most cases of acute acquired toxoplasmosis (AAT) are oligosymptomatic and self-limited. Therefore, these infections rarely indicate treatment. Prospective studies of AAT patients are rare in the medical literature. The frequency of systemic manifestations has not been sufficiently studied. In order to search for risks factors for systemic and ocular involvement, 37 patients were submitted to a diagnostic investigative protocol. The most frequent findings were lymph node enlargement (94.6%), asthenia (86.5%), headache (70.3%), fever (67.6%) and weight loss (62.2%). Hepatomegaly and/or splenomegaly were present in 21.6% of cases (8/37). Liver transaminases were elevated in 11 patients (29.7%) and lactic dehydrogenase in 17 patients (45.9%). Anaemia was found in four patients (10.8%), leucopoenia in six patients (16.2%), lymphocytosis in 14 patients (37.8%) and thrombocytopenia in one patient (2.7%). Fundoscopic examination revealed retinochoroiditis in four patients (10.8%). No statistical association was found between any one morbidity and retinochoroiditis. Nevertheless, a significant association was found between the presence of more than eight morbidity features at evaluation and long-lasting disease. An ideal diagnostic protocol for AAT would include evidence of systemic involvement. Such a protocol could be used when planning treatment.

Acute acquired toxoplasmosis: clinical-laboratorial aspects and ophthalmologic evaluation in a cohort of immunocompetent patients

Most cases of acute acquired toxoplasmosis (AAT) are oligosymptomatic and self-limited. Therefore, these infections rarely indicate treatment. Prospective studies of AAT patients are rare in the medical literature. The frequency of systemic manifestations has not been sufficiently studied. In order to search for risks factors for systemic and ocular involvement, 37 patients were submitted to a diagnostic investigative protocol. The most frequent findings were lymph node enlargement (94.6 percent), asthenia (86.5 percent), headache (70.3 percent), fever (67.6 percent) and weight loss (62.2 percent). Hepatomegaly and/or splenomegaly were present in 21.6 percent of cases (8/37). Liver transaminases were elevated in 11 patients (29.7 percent) and lactic dehydrogenase in 17 patients (45.9 percent). Anaemia was found in four patients (10.8 percent), leucopoenia in six patients (16.2 percent), lymphocytosis in 14 patients (37.8 percent) and thrombocytopenia in one patient (2.7 percent). Fundoscopic examination revealed retinochoroiditis in four patients (10.8 percent). No statistical association was found between any one morbidity and retinochoroiditis. Nevertheless, a significant association was found between the presence of more than eight morbidity features at evaluation and long-lasting disease. An ideal diagnostic protocol for AAT would include evidence of systemic involvement. Such a protocol could be used when planning treatment.

Genotyping of Giardia duodenalis from Southern Brown Howler Monkeys (Alouatta clamitans) from Brazil.

Giardia duodenalis is a widespread intestinal protozoan that can infect humans and animals, both domestic and wild. Independent of host, infections present with the same symptoms. However, based on host specificity, Giardia isolates have been grouped into genotypes A to G. Parasites of assemblage A and B are known to infect humans, in addition to primates and a wide variety of mammals. In Brazil, hitherto Giardia genotypes were defined only for humans and domestic animals. To evaluate the genotypes of different Giardia present among other animals, fecal samples from 28 Southern Brown Howler Monkeys (Alouatta clamitans) kept in captivity from South Brazil were screened for G. duodenalis using parasitological methods. All of them were asymptomatic, but positive for Giardia. The genotype of the G. duodenalis circulating among these animals was ascertained by molecular typing, performed using amplification and sequencing of the beta-giardin gene. Sixteen of 28 samples were successfully amplified by PCR and sequencing of this gene s revealed that all of them were of the genotype A1. These findings suggest that A. clamitans represent a potential risk of environmental contamination of a G. duodenalis genotype that also infect humans, and therefore can be considered a potential reservoir for G. duodenalis of a genotype that can also infects humans. Therefore, these results highlight a potential public health problem due to the epidemiological and molecular evidence for anthropozoonotic transmission.

Genotyping of Giardia duodenalis from human and animal samples from Brazil using beta-giardin gene: a phylogenetic analysis.

Giardia duodenalis is one of the major diarrhea agents in human and animals distributed worldwide, and present high levels of genetic diversity, showing seven genotypes: A, B, C, D, E, F, and G. Only Assemblages A and B have been detected in humans and in a wide range of other mammalians hosts, whereas the remaining Assemblages (C-G) are host-specific. Molecular characterization of cysts of human and animal origin are useful to address the co-circulate isolates between these host, and represents an objective means to evaluate zoonotic infection hypothesis. In the present work the G. duodenalis genotypes were characterized by restriction fragment length polymorphisms and DNA sequencing analysis of PCR products of the beta-giardin gene. The cysts were collected in the city of Rio de Janeiro, in Brazil, from a population composed by humans (n=366, 310 children and 56 adults), domestic animals (n=11) from a municipal daycare center in the surroundings of a slum and neighborhood medium-high class domestic animals (n=18). Parasitological exams were developed in human fecal samples. Parasites were found in 60% (186/310) and 66% (37/56) of the samples from children and adults, respectively. Among children's samples, 27.7% (86/310) were positive for G. duodenalis. Only 1.7% (1/56) of the adults was positive for this parasite. In general a total of 87 fecal samples (86 from children and 1 from adult) from all population studied were positive for G. duodenalis, and 62 of these were subjected to molecular analysis using a PCR that amplified a fragment of the beta-giardin gene. Sixty samples were typed as genotype A1, two as genotype A2 and genotype B was not encountered. Among domestic animals samples (n=29), eight (seven dogs and one cat) from the slum community were identified as genotype A1, and all control samples (n=18) were negative in the molecular assay. The host-specific genotypes C, D and, F were not found. In this study we described single case of G. duodenalis infection associated with a child and her dog and both isolates characterized as genotype A1. Despite the low incidence, this data suggest the putative existence of a zoonotic cycle of G. duodenalis in the studied population.

Mapping of the distribution of Trypanosoma cruzi infection among small wild mammals in a conservation unit and its surroundings (Northeast-Brazil).

Maps are a useful tool that permits correlation of landscapes with hotspots of parasite transmission. Here, they were used as a tool for geovisualization to evaluate variables involved in the transmission of Trypanosoma cruzi among small wild mammals in an area endemic for Chagas disease, the "Serra da Capivara" National Park (PARNA) and its surroundings in Piauí State, Northeast Brazil. The implementation of a Geographical Information System (GIS) allowed the observation that a previously noted aggregated distribution of Triatoma sordida and Triatoma brasiliensis, T. cruzi prevalence and infection pattern of small wild mammals was directly or indirectly influenced by the local relief and human action. Small mammalian species diversity was higher in mesic refugia inside the park and in its buffer zone and lower in the disturbed area by anthropic activities. Didelphis albiventris was more abundant in the areas affected by human action. Thrichomys laurentius demonstrated to be an eclectic species and a competent reservoir of T. cruzi, being infected in all study areas. Small wild mammals infected with the TCII genotype of T. cruzi were localized only in the buffer zone of PARNA while TCI infected specimens were found in both areas, inside the PARNA and its buffer zone. The impact of biodiversity loss on the transmission cycle of T. cruzi in the wild environment was discussed.

Stable infection of primates with Trypanosoma cruzi I and II.

In order to better comprehend the putative association between genotype Trypanosoma cruzi II and primates, an evaluation of the infection in free ranging primates and specimens born in captivity from different geographical areas, the Amazon and the Atlantic forest, was carried out. Seroprevalences of the T. cruzi infection among the primates was similar in both biomes (45.5% and 46%). The parasites were isolated from 8 and 4 different species of primates, respectively from the Amazon and Atlantic forest. Multi-locus enzyme electrophoresis (MLEE) typed the isolates from Amazon as zymodeme 1. Mini-exon gene analysis characterized all these isolates as T. cruzi I, the main genotype circulating in the region. In the Atlantic forest, primates infected with TCI and TCII, as well as a mixed infection (TCI and TCII), were detected. These findings prove that primates may maintain stable infections by both genotypes. Moreover, data show that T. cruzi can occur in a wide range of primate genera, independent of their social behaviour, niches or habitats. Considering the high seroprevalence and stability of T. cruzi infection among the primates, these animals play an important role in the maintenance of the parasite in nature.

A zymographic study of metalloprotease activities in extracts and extracellular secretions of Leishmania (Viannia) braziliensis strains.

Proteolytic activities of 5 strains of Leishmania (Viannia) braziliensis isolated from Brazilian and Colombian patients, presenting distinct clinical manifestations, were characterized and compared using whole-promastigote extracts and extracellular secretions. Zymographic assays concerning whole-cell extracts and supernatants resulted in the detection of high molecular weight bands, ranging from 50 to 125 kDa. Proteolytic activities from both whole-cell extracts and supernatants were optimal in a pH range 5.5 to 9.0 for all analysed strains. Such protease activities were inhibited when 10 mM 1,10-phenanthroline was assayed, strongly suggesting that the enzymes responsible for hydrolysis of the substrate belong to the metalloproteases class. Distinct profiles of metalloproteases were observed among the studied L. (V.) braziliensis strains. Differences among the microorganisms might be related to the geographical origin of the strains and/or to the clinical presentation.