Identification of a phylogenetically distinct orthobunyavirus from group C.

Apeu virus (APEUV) (family Bunyaviridae, genus Orthobunyavirus) was plaque purified and characterised by serological and molecular analysis. Neutralising assays confirmed cross-reactivity between purified APEUV clones and the Caraparu virus complex of group C orthobunyaviruses. Partial sequencing of the L, M and S segments of one APEUV clone (APEUV-CL5) was carried out. A phylogenetic tree constructed with the L amino acid sequences clustered APEUV-CL5 within the genus Orthobunyavirus, confirming its serological classification. Analysis of M segment sequences clustered APEUV-CL5 in the Caraparu virus complex (Group C), in agreement with serological tests and previous molecular characterisation. However, the sequence of the nucleocapsid gene (N) gave low identity values when compared to those of the group C viruses. The phylogenetic tree based on N nucleotide sequences clustered APEUV-CL5 next to the California and Bwamba groups. This remarkable S nucleotide variability suggests that APEUV-CL5 could be a genetic reassortant and that this evolutionary mechanism is present in the history of the group C viruses.

A-type inclusion bodies: a factor influencing cowpox virus lesion pathogenesis.

The family Poxviridae comprises the most complex animal DNA viruses. During some poxvirus infections, A-type inclusion bodies (ATIs), codified by the ati gene, are produced. Although some studies have compared poxviruses that encode these inclusion bodies with those that do not, the biological function of ATIs is poorly understood. A recombinant ati-deleted cowpox virus was constructed and compared with the wild-type virus in in vitro experiments including electron microscopy and plaque and viral growth assays. No significant differences were observed in vitro. This reinforces the conclusion that the inclusion body is not essential for in vitro viral replication and morphogenesis. Additionally, different lesion progressions in vivo were observed by macroscopic and histological analysis, suggesting that the presence or absence of ATIs could result in different healing dynamics. This is the first time that the role of ATIs during viral replication has been studied based solely on one variable, the presence or absence of ATIs.

Natural human infections with Vaccinia virus during bovine vaccinia outbreaks.

BACKGROUND: Bovine vaccinia is an exanthematic disease caused by Vaccinia virus (VACV). This zoonosis has been associated with several cases of bovine infection, particularly in milk herds. Farmers, milkers and their close contacts developed lesions on the hands, forearms, legs and face accompanied by fever, headache, malaise, myalgia and axillary, inguinal and cervical lymphadenopathy. VACV infections have a significant public health impact due to their occupational character, high frequency of transmission and the improper medical treatment often applied. OBJECTIVES: To study natural human infection by VACV and to analyze clinical and epidemiological aspects, emphasizing the patients' immunological status. STUDY DESIGN: Ninety-eight individuals from rural properties with bovine vaccinia (BV) outbreaks who were at risk due to contact were submitted to epidemiological and clinical studies. From these individuals, 54 sera were analyzed by serological and molecular procedures. This study was conducted in Rio de Janeiro State from September 2002 to October 2006. RESULTS: The clinical frequency of infection was 52.0%, with 57.4% ELISA and 43.0% PRNT-positive reactions. DNAemia was detected in 18.5% of the analyzed sera, and 50% of smallpox-vaccinated individuals developed symptoms. CONCLUSIONS: This study confirms the high clinical frequency of human VACV infection, even among vaccinated individuals. The infection was related to detection of IgG- or IgM-specific antibodies that correlates in most of the cases with positive PRNT. The DNAemia suggests viremia during VACV natural infections. Our data indicate that patients vaccinated against smallpox may no longer be protected.

Sequence and phylogenetic analysis of the large (L) segment of the Tahyna virus genome.

The Tahyna virus (TAHV) is an important human pathogen in the Bunyaviridae family. To date, only the S and M segments of this virus have been sequenced, but the sequence of the L segment hasn't been established yet. In this study, we sequenced 963 nucleotides of the L segment of TAHV, comprising pre-motif A and motif A in region 3 of the RNA polymerase gene.

Caraparu virus (group C Orthobunyavirus): sequencing and phylogenetic analysis based on the conserved region 3 of the RNA polymerase gene.

Here, for the first time, we report the nucleotide sequence of Caraparu virus (CARV) L segment and the analysis of the RNA polymerase region 3 encoded by this segment. The 1,404 bp nucleotide sequence shares the highest identity with Bunyamwera, La Crosse, Oropouche, and Akabane virus sequences. The amino acid sequence was deduced and aligned with sequences from members of the Bunyaviridae family and used for phylogenetic analysis. The CARV clustered in the Orthobunyavirus genus. The premotif A and motifs A-E are present in the region 3 of the Bunyaviridae family, were also conserved in CARV L protein, as well as other conserved regions among Orthobunyavirus genus.