Caracterization of Aujeszky's disease virus isolated from South Brazil in the last twenty years by restriction enzyme analysis

Abstract

Aujeszky's disease virus (ADV) belongs to Herpesviridae family and is an important etiological agent which infects pigs causing economic losses in swine producing countries worldwide and international trade restrictions to products of swine origin. An eradication program for ADV was established in Santa Catarina State since 2001. The last outbreak was reported in July 2004 and since then none has been reported. The disease has been controlled with the use of a genetic modified vaccine and elimination of seropositives. Aiming the characterization of ADV isolated in the South of Brazil in the last twenty years , a retrospective study based on the genomic analysis of the isolates through a digestion of viral genomic DNA with restriction enzyme Bam HI was done. Thirty-seven ADV samples isolated from swine from the States of Santa Catarina, Parana and Rio Grande do Sul were analyzed. These isolates were compared to the reference strains NIA-4, Bartha and Begonia. The most predominant genomic arrangement was type II found in 33 samples isolated in Santa Catarina State and in one isolate from Rio Grande do Sul State. Genomic arrangement type I, characteristic of vaccine strains was identified in 2 isolates from Parana State and in 1 isolate from Rio Grande do Sul State.

Principais ameaças sanitárias endêmicas da cadeia produtiva de suínos no Brasil

“…A doença de Aujeszky tem sido relatada no rebanho brasileiro desde 1912, e, em determinadas regiões, ocorre de forma esporádica (Schaefer et al, 2006;Ciacci-Zanella et al, 2008). No Brasil, a vacina contra essa doença é permitida, porém seu uso é controlado pelo Ministério da Agricultura, Pecuária e Abastecimento (Mapa).…”

Cloning and expression of Aujeszky's disease virus glycoprotein E (gE) in a baculovirus system

“…Genomic DNA viral samples, 0261/83 (Embrapa), were digested with BamHI restriction enzyme and presented the genotype II pattern (19). The digestion revealed the presence of fragment BamHI 7, which corresponds to a part of US region (unique short) of the ADV genome fragment 7, in which the gE gene is located (9,10).…”

“…After 18 hours, the culture supernatant was harvested and centrifuged on a sucrose cushion 25% at 100,000 x g for 3 hours at 4ºC. The viral pellet was re-suspended in TNE (Tris-Cl 10 mM, pH 7,4; NaCl 150 mM; EDTA 1 mM, pH 8.0) and treated with 1% sodium dodecyl sulfate (SDS) and 20 µg/µL proteinase K for 1 h at 37ºC (19). After digestion, the viral DNA was extracted with phenol, precipitated with cold ethanol, re-suspended in TE pH 8.0 and maintained at 4ºC (18).…”

“…The gE is a non essential protein for viral replication (5,(19)(20)(21) and the deletion of the gE gene has been used for the production of marker vaccines (4,13). The immunization strategies are important to control the disease by reducing signs and viral spread, and for differentiation of vaccinated from naturally infected animals.…”

Diagnóstico e genotipagem do vírus da pseudoraiva por nested-PCR e análise de restrição enzimática

“…Análises de RFLP permitiram detectar que este era o mesmo tipo genômico presente no Estado de Santa Catarina, que ainda não havia completado seu programa de erradicação na época do surto no RS (SILVA et al, 2005). O mesmo método identificou amostras do genótipo I no Estado do Paraná, um tipo incomum no Brasil, mas presente na Argentina, o que indica a possível introdução desse genótipo por animais com o vírus latente daquele país (SCHAEFER et al, 2006). O método de análise por restrição enzimática de genoma completo com BamHI é útil na diferenciação do PrV, mas muito complexo, pois requer isolamento do vírus, processos de ultracentrifugação para purificação do DNA e eletroforese de temperatura controlada que pode durar até 20 horas.…”