A Retrospective Overview of Enterovirus Infection Diagnosis and Molecular Epidemiology in the Public Hospitals of Marseille, France (1985–2005)

Human enteroviruses (HEV) are frequent human pathogens and, associated in particular with large outbreaks of aseptic meningitis. Here, we have compiled a database of clinical HEV isolates from the Public Hospitals of Marseille, from 1985 to 2005. Amongst 654 isolates that could be characterized by complete sequencing of the VP1 gene, 98% belonged to species HEV-B; the most frequently isolated serotypes were Echovirus E30, E11, E7, E6 and E4. The high incidence of E30 and the recent emergence of E13 are consistent with reports worldwide and peak HEV isolation occurred mostly in the late spring and summer months. The proportion of echoviruses has decreased across the years, while that of coxsackieviruses has increased. Stool (the most frequent sample type) allowed detection of all identified serotypes. MRC5 (Human lung fibroblasts) cell line was the most conducive cell line for HEV isolation (84.9% of 10 most common serotype isolates, 96.3% in association with BGM (Buffalo green monkey kidney cells)). Previous seroneutralization-based serotype identification demonstrated 55.4% accuracy when compared with molecular VP1 analysis. Our analysis of a large number of clinical strains over 20 years reinforced the validity of VP1 serotyping and showed that comparative p-distance scores can be coupled with phylogenetic analysis to provide non-ambiguous serotype identification. Phylogenetic analysis in the VP1, 2C and 3D regions also provided evidence for recombination events amongst clinical isolates. In particular, it identified isolates with dissimilar VP1 but almost identical nonstructural regions

Leishmaniose tegumentar americana canina em municípios do norte do Estado do Paraná, Brasil Canine American cutaneous leishmaniasis in municipalities of northern Paraná State, Brazil

A leishmaniose tegumentar americana (LTA) é endêmica no norte do Estado do Paraná, Brasil, afetando tanto o homem como os cães. O objetivo deste trabalho foi estudar a ocorrência da doença na população canina em áreas endêmicas de LTA. No período de setembro de 1999 a julho de 2002, foram investigados cães em sete localidades rurais de cinco municípios onde ocorreram casos humanos autóctones da doença, utilizando métodos parasitológicos e sorológicos. Dos 67 cães estudados, 14 (20,9%) tinham lesões sugestivas de LTA, dos quais 3 (21,4%) estavam infectados por Leishmania sp. Trinta e sete (55,2%) cães apresentaram imunofluorescência indireta positiva para Leishmania (Viannia) braziliensis. Os resultados mostram a ocorrência simultânea de leishmaniose humana e canina e indicam a necessidade de estudos adicionais para esclarecer o papel do cão no ciclo de transmissão do parasito nessas áreas.<br>American cutaneous leishmaniasis (ACL) is an endemic disease in northern Paraná State, where it affects humans and dogs. This study aimed to verify the occurrence of the canine disease in ACL endemic areas. From September 1999 to July 2002, dogs were investigated in 7 rural areas of 5 municipalities where autochthonous human cases of ACL had been reported. Parasitological and serological methods were used. Fourteen of 67 dogs studied (20.9%) had lesions suggestive of ACL, of which 3 (21.4%) were infected with Leishmania sp. Indirect immunofluorescence for Leishmania (Viannia) braziliensis was positive in 37 (55.2%) of 67 dogs. The results showed that human and canine ACL occur simultaneously and indicate the need for additional studies to elucidate the role of dogs in the ACL transmission cycle in the area studied

Echovirus 30 associated with cases of aseptic meningitis in state of Pará, Northern Brazil

Investigation of the aetiology of viral meningitis in Brazil is most often restricted to cases that occur in the Southern and Southeastern Regions; therefore, the purpose of this study is to describe the viral meningitis cases that occurred in state of Pará, Northern Brazil, from January 2005-December 2006. The detection of enterovirus (EV) in cerebrospinal fluid was performed using cell culture techniques, RT-PCR, nested PCR and nucleotide sequencing. The ages of the 91 patients ranged from < one year old to > 60 years old (median age 15.90 years). Fever (87.1%), headache (77.0%), vomiting (61.5%) and stiffness (61.5%) were the most frequent symptoms. Of 91 samples analyzed, 18 (19.8%) were positive for EV. Twelve were detected only by RT- PCR followed by nested PCR, whereas six were found by both cell culture and RT-PCR. From the last group, five were sequenced and classified as echovirus 30 (Echo 30). Phylogenetic analyses revealed that Echo 30 detected in Northern Brazil clustered within a unique group with a bootstrap value of 100% and could constitute a new subgroup (4c) according to the phylogenetic tree described by Oberste et al. (1999). This study described the first molecular characterization of Echo 30 in Brazil and this will certainly contribute to future molecular analyses involving strains detected in other regions of Brazil