Comparative performance evaluation of hepatitis C virus genotyping based on the 5' untranslated region versus partial sequencing of the NS5B region of brazilian patients with chronic hepatitis C

Background Genotyping of hepatitis C virus (HCV) has become an essential tool for prognosis and prediction of treatment duration. The aim of this study was to compare two HCV genotyping methods: reverse hybridization line probe assay (LiPA v.1) and partial sequencing of the NS5B region. Methods Plasma of 171 patients with chronic hepatitis C were screened using both a commercial method (LiPA HCV Versant, Siemens, Tarrytown, NY, USA) and different primers targeting the NS5B region for PCR amplification and sequencing analysis. Results Comparison of the HCV genotyping methods showed no difference in the classification at the genotype level. However, a total of 82/171 samples (47.9%) including misclassification, non-subtypable, discrepant and inconclusive results were not classified by LiPA at the subtype level but could be discriminated by NS5B sequencing. Of these samples, 34 samples of genotype 1a and 6 samples of genotype 1b were classified at the subtype level using sequencing of NS5B. Conclusions Sequence analysis of NS5B for genotyping HCV provides precise genotype and subtype identification and an accurate epidemiological representation of circulating viral strains.

Identification of SL addition trans-splicing acceptor sites in the internal transcribed spacer I region of pre-rRNA in Leishmania (Leishmania) amazonensis

Trypanosomatidae is a family of early branching eukaryotes harbouring a distinctive repertoire of gene expression strategies. Functional mature messenger RNA is generated via the trans-splicing and polyadenylation processing of constitutively transcribed polycistronic units. Recently, trans-splicing of pre-small subunit ribosomal RNA in the 5' external transcribed spacer region and of precursor tRNAsec have been described. Here, we used a previously validated semi-nested reverse transcription-polymerase chain reaction strategy to investigate internal transcribed spacer (ITS) I acceptor sites in total RNA from Leishmania (Leishmania) amazonensis. Two distinct spliced leader-containing RNAs were detected indicating that trans-splicing reactions occur at two AG acceptor sites mapped in this ITS region. These data provide further evidence of the wide spectrum of RNA molecules that act as trans-splicing acceptors in trypanosomatids.

Relations between Depression, Alcohol and Gender in the Metropolitan Region of São Paulo, Brazil

As part of the GENACIS project, this paper sought to assess the prevalence of depression in an urban sample in the city of São Paulo, Brazil, as well as the association between depression and alcohol abuse according to gender. To achieve this, an epidemiological survey was conducted using a stratified probability sample, including 2,083 adults. CIDI SF was used to identify depression. The Rao Scott test and multivariate logistic regression were used for statistical analysis. The response rate was 74.9%. Females (58.3%) under 40 years of age (52%) were predominant. The prevalence of depression was 28.3% for women and 12.7% for men. Most men declared being drinkers (61.1%) in the last year. Depression was associated with an alcohol drinking pattern, mostly binge drinking, in addition to the occurrence of problems derived from alcohol use. Most women declared being abstainers (69.5%). Depression was associated with cohabiting with spouses with alcohol-related problems. Results reveal that the association between depression and alcohol consumption is distinct between genders.

Molecular epidemiology of animal rabies in the semiarid region of Paraíba, Northeastern Brazil

In the semiarid of the state of Paraíba, the anti-rabies vaccination is not common, most of the local inhabitants who deal with the animals do not know the incidence of the disease in the region. In this study, samples of foxes (Pseudalopex vetulus), insectivorous bats (Molossus molossus), raccoons (Procyon cancrivorous) and domestic animals brains were submitted to the diagnosis of rabies, by using the direct fluorescent antibody technique (d-FAT) and mouse inoculation test (MIT). Of the 581 examined materials, 50 (8.60 %) were positive for d-FAT and 47 (8.09 %) for MIT. From the positive samples for rabies, RNAs were extracted and transformed to cDNA, at the Laboratory of Rabies/Faculdade de Medicina Veterinária e Zootecnia/USP, SP. The phylogenetic characterization of the N gene was performed at the Universidade de São Paulo, Faculdade de Medicina Veterinária e Zootecnia, Departamento de Medicina Veterinária Preventiva e Saúde Animal, Universidade Nihon, Faculdade de Ciências Bioresource, Fujisawa, Kanagawa, Japão. Based on the results of genotyping and phylogenetic analyzes, it is concluded that the epidemiology of rabies is complex in the semiarid of Paraíba, with different viral variants being maintained in domestic dogs, foxes, insectivorous bats and vampire bats. All the isolates examined belong to the genotype I of the genus Lyssavirus and it is possible to state that in the region, foxes are important sylvatic reservoirs of the rabies virus.

Serological survey on Ehrlichia sp. among dogs in the central region of Rio Grande do Sul

A serological survey on Ehrlichia canis was conducted among dogs in the central area of the state of Rio Grande do Sul, where the tick Rhipicephalus sanguineus is a common parasite of dogs. Out of a total of 316 dogs attended at the veterinary teaching hospital in the municipality of Santa Maria, only 14 (4.43%) reacted positively to E. canis antigens in the indirect immunofluorescence assay, with the following endpoint titers: 80 (three dogs), 160 (five), 320 (four), 640 (one) and 1280 (one). Like in previous studies in other regions of the state of Rio Grande do Sul, only a very small portion of the dogs in Santa Maria presented antibodies reactive to E. canis, even though canine infestations due to R. sanguineus are very common in this study region. These results contrast with other regions of Brazil, where E. canis is endemic among canine populations, with seropositivity values generally higher than 30%. Genetic differences among the R. sanguineus populations in South America might be implicated in these contrasting results.

Mycobacterium bovis infection in goats from the Northeast Region of Brazil

A total of 8,058 male and female mixed-breed goats and 1-4 years of age were slaughtered over a period of 7 months at the public slaughterhouse of Patos city, Paraíba state, in the Northeast region of Brazil; 822 animals were inspected for gross lesions of tuberculosis, and 12 (1.46%) had lesions suggestive of tuberculosis in the mammary gland, lungs, liver and mediastinal, mesenteric, submandibular, parotid and prescapular lymph nodes. Presence of granulomatous lesions was confirmed in the submandibular lymph node of one (8.3%) goat at the histopathological examination and at the mycobacterium culture the same sample was confirmed positive. Isolate was confirmed as belonging to the M. tuberculosis complex by PCR restriction enzyme analysis (PRA). Spoligotyping identified the isolate into spoligotype SB0295 on the M. bovis Spoligotype Database website (www.mbovis.org), and it was classified as M. bovis. The occurrence of M. bovis in goats in this study suggests that this species may be a potential source of infection for humans and should be regarded as a possible problem in the advancement of control and eradication program for bovine tuberculosis in Brazil.

Paleoenvironmental reconstruction of a Late Quaternary lagoon system in southern Brazil (Jaguaruna region, Santa Catarina state) based on multi-proxy analysis

This paper provides a paleoenvironmental reconstruction of a Late Quaternary lagoon system in the Jaguaruna region of Santa Catarina state, southern Brazil. Integrated results of bulk sedimentary organic matter characterization (delta C-13, delta N-15 and C/N), microfossil (pollen and diatom) and grain-size analysis from three shallow cores (similar to 2.5m depth) allowed us to propose an evolving paleogeographic scenario in this coastal region for the last ca. 5500 cal a BP. The lagoonal system in this area was more extensive during the mid-Holocene than today, with a gradual and continuous lagoon-sea disconnection until the present. We add to the debate regarding relative sea-level (RSL) variations for the Brazilian coast during the Holocene and discuss the importance of sedimentary dynamics for interpreting changes in coastal ecosystems. The multi-proxy analysis suggests that changes in coastal ecosystems could be directly related to local sedimentary processes, which are not necessarily linked to RSL fluctuations and/or to climatic variations. Copyright (c) 2011 John Wiley & Sons, Ltd.

Sero-epidemiological evaluation of changes in Plasmodium falciparum and Plasmodium vivax transmission patterns over the rainy season in Cambodia

Background: In Cambodia, malaria transmission is low and most cases occur in forested areas. Seroepidemiological techniques can be used to identify both areas of ongoing transmission and high-risk groups to be targeted by control interventions. This study utilizes repeated cross-sectional data to assess the risk of being malaria sero-positive at two consecutive time points during the rainy season and investigates who is most likely to sero-convert over the transmission season. Methods: In 2005, two cross-sectional surveys, one in the middle and the other at the end of the malaria transmission season, were carried out in two ecologically distinct regions in Cambodia. Parasitological and serological data were collected in four districts. Antibodies to Plasmodium falciparum Glutamate Rich Protein (GLURP) and Plasmodium vivax Merozoite Surface Protein-119 (MSP-119) were detected using Enzyme Linked Immunosorbent Assay (ELISA). The force of infection was estimated using a simple catalytic model fitted using maximum likelihood methods. Risks for sero-converting during the rainy season were analysed using the Classification and Regression Tree (CART) method. Results: A total of 804 individuals participating in both surveys were analysed. The overall parasite prevalence was low (4.6% and 2.0% for P. falciparum and 7.9% and 6.0% for P. vivax in August and November respectively). P. falciparum force of infection was higher in the eastern region and increased between August and November, whilst P. vivax force of infection was higher in the western region and remained similar in both surveys. In the western region, malaria transmission changed very little across the season (for both species). CART analysis for P. falciparum in the east highlighted age, ethnicity, village of residence and forest work as important predictors for malaria exposure during the rainy season. Adults were more likely to increase their antibody responses to P. falciparum during the transmission season than children, whilst members of the Charay ethnic group demonstrated the largest increases. Discussion: In areas of low transmission intensity, such as in Cambodia, the analysis of longitudinal serological data enables a sensitive evaluation of transmission dynamics. Consecutive serological surveys allow an insight into spatio-temporal patterns of malaria transmission. The use of CART enabled multiple interactions to be accounted for simultaneously and permitted risk factors for exposure to be clearly identified.

Survey of Bancroftian filariasis infection in humans and Culex mosquitoes in the western Brazilian Amazon region: implications for transmission and control

Introduction: The aim of this work was to identify possible lymphatic filariasis foci in the western Brazilian Amazonian that could be established from the reports of Rachou in the 1950s. The study was conducted in three cities of the western Brazilian Amazon region - Porto Velho and Guajará-Mirim (State of Rondônia) and Humaitá (State of Amazonas). Methods: For human infection evaluation thick blood smear stained with Giemsa was used to analyze samples collected from 10pm to 1am. Polymerase chain reaction (PCR) was used to examine mosquito vectors for the presence of Wuchereria bancrofti DNA. Humans were randomly sampled from night schools students and from inhabitants in neighborhoods lacking sanitation. Mosquitoes were collected from residences only. Results: A total 2,709 night students enrolled in the Program for Education of Young Adults (EJA), and 935 people registered in the residences near the schools were examined, being 641 from Porto Velho, 214 from Guajará-Mirim and 80 from Humaitá. No individual examined was positive for the presence of microfilariae in the blood stream. A total of 7,860 female Culex quinquefasciatus specimens examined were negative by PCR. Conclusions: This survey including human and mosquito examinations indicates that the western Amazon region of Brazil is not a focus of Bancroftian filariasis infection or transmission. Therefore, there is no need to be included in the Brazilian lymphatic filariasis control program.