Spread of poxviruses in livestock in Brazil associated with cases of double and triple infection.

The objective of this work is to describe the distribution of outbreaks of vaccinia virus (VACV), pseudocowpox virus (PCPV), and bovine papular stomatitis virus (BSPV) in Brazil. The Official Laboratory of the Brazilian Ministry of Agriculture received 89 samples from different locations in Brazil in 2015 and 2016 for diagnosis of vesicular and exanthematous disease. Poxvirus coinfections occurred in 11 out of 33 outbreaks, including the first reported triple infection by BPSV, PCPV, and VACV. This occurrence may be associated with the circulation of these viruses in Brazilian cattle.

Detection of pseudocowpox virus in water buffalo (Bubalus bubalis) with vesicular disease in the state of São Paulo, Brazil, in 2016.

BACKGROUND: Parapoxviruses are zoonotic viruses that infect cattle, goats and sheep; there have also been reports of infections in camels, domestic cats and seals. OBJECTIVE: The objective of this report was to describe a case of vesicular disease caused by pseudocowpox virus (PCPV) in water buffalo (Bubalus bubalis) in Brazil. ANIMALS: Sixty buffalo less than 6 months old exhibited ulcers and widespread peeling of the tongue epithelium. There were no cases of vesicular disease in pigs or horses on the same property. METHODS: Samples were analysed by PCR and sequencing. Phylogenetic analysis in MEGA 7.01 was reconstructed using major envelope protein (B2L) by the Tamura three-parameter nucleotide substitution model and the maximum likelihood and neighbor joining models, both with 1000 bootstrap replicates. The genetic distance between the groups was analysed in MEGA using the maximum composite likelihood model. The rate variation among sites was modeled using gamma distribution. RESULTS: The presence of PCPV in the buffalo herd could be demonstrated in epithelium and serum. The minimum genetic distance between the isolated PCPV strain (262-2016) and orf virus and bovine papular stomatitis virus was 6.7% and 18.4%, respectively. The maximum genetic distance calculated was 4.6% when compared with a PCPV detected in a camel. Conclusions/Clinical Importance: The peculiar position of the isolated strain in the phylogenetic trees does not necessarily indicate a different kind of PCPV that infects buffalo. More samples from cattle and buffalo in Brazil must be sequenced and compared to verify if PCPV from buffalo are genetically different from samples derived from cattle.

Comparison of nine DNA extraction methods for the diagnosis of bovine tuberculosis by real time PCR / Comparação de nove métodos de extração de DNA para diagnóstico de tuberculose bovina por PCR em tempo real

ABSTRACT: Bovine tuberculosis is an infectious disease with a high impact on the cattle industry, particularly in developing countries. PCR is a very sensitive method for detection of infectious agents, but the sensitivity of molecular diagnosis is largely dependent on the efficiency of the DNA extraction methods. The objective of this study was to evaluate DNA extraction methods for direct detection of Mycobacterium bovis in bovine tissue. Nine commercial kits for DNA extraction were evaluated when combined with two real time PCRs. The DNeasy Blood & Tissue Kit from QIAGEN showed better performance and sensitivity followed by the DNA Mini Kit RBC and FTA Elute Micro Card. Results suggested that, even when the analytical sensitivity of the qPCR is very high, the extraction method can influence the diagnostic sensitivity.

RESUMO: A tuberculose bovina é uma doença infecciosa com um alto impacto na pecuária, particularmente em países em desenvolvimento. A PCR é um método muito sensível para a detecção de agentes infecciosos, mas a sensibilidade do diagnóstico molecular é em grande parte dependente da eficiência dos métodos de extração de DNA. O objetivo deste estudo foi avaliar métodos de extração de DNA para detecção direta de Mycobacterium bovisem tecido bovino. Nove kits comerciais para extração de DNA foram avaliados, quando combinados com duas PCRs em tempo real. O Kit Dneasy Blood & Tissue da Qiagen apresentou melhor desempenho e sensibilidade, seguido dos kits DNA Mini RBC e FTA Elute Micro Card (protocolo modificado com digestão enzimática prévia). Os resultados sugerem que, mesmo quando a sensibilidade analítica do qPCR é muito elevada, o método de extração pode influenciar na sensibilidade de diagnóstico.

Avaliação de contaminantes inorgânicos (As, Cd, Cr, Hg e Pb) em espécies de peixes / Inorganic contaminants (As, Cd, Cr, Hg and Pb) in fish species

O teor dos contaminantes arsênio total, cádmio, crômio, chumbo e mercúrio total, foi avaliado em 240 amostras das espécies de peixes pescada (Macrodon ancylodon), tainha (Mugil liza), corvina (Micropogoniasfurnieri) e sardinha (Sardinella brasiliensis), adquiridas no comércio atacadista da Companhia de Entrepostos e Armazéns Gerais de São Paulo (CEAGESP), no inverno de 2009 e no verão de 2010. As determinações dos contaminantes inorgânicos foram realizadas por espectrometria de emissão óptica em plasma com acoplamento indutivo (ICP OES). Os intervalos de concentração obtidos, em mg.kg-1(peso úmido), foram de As total (<0,1-8,63); Cd (<0,01-0,287); Cr (<0,02-0,44); Pb (<0,02-2,92) e Hg total(<0,002-0,285). As espécies mais contaminadas foram a sardinha e a corvina, sendo encontrados As e Cr em níveis acima dos limites máximos permitidos pela legislação brasileira (1 e 0,1 mg kg-1), em 67 por cento e 7 por cento das amostras analisadas, respectivamente. Este estudo demonstra a ocorrência de contaminação por arsênio, crômio e chumbo nas espécies de pescada, tainha, corvina e sardinha. O maior índice de arsênio foi detectado na espécie sardinha. Na corvina, os contaminantes As, Cr e Pb foram encontrados em teores mais elevados no período de inverno do que na época de verão.

Lipopolysaccharide enhances glutaric acid-induced seizure susceptibility in rat pups: behavioral and electroencephalographic approach.

Glutaric acidemia type I (GA-I) is an inherited metabolic disease characterized by accumulation of glutaric acid (GA) and seizures. Considering that seizures are precipitated by common infections in children with GA-I, we investigated whether lipopolysaccharide (LPS) modifies GA-induced electrographic and neurochemical alterations in 21 days-old rats. The effect of LPS on convulsive behavior and electroencephalographic (EEG) alterations induced by GA (0.13; 0.4; 1.3 µmol/striatum) was determined in freely moving rats. After EEG recordings, we measured the levels of interleukin 1ß (IL-1ß) in GA-injected striatum. The injection of LPS (2mg/kg; i.p.) 6h before of GA administration, reduced the latency and increased the duration of seizures induced by GA (1.3 µmol/site). In addition, LPS administration increased IL-1ß striatal levels, which positively correlated with total time in seizures. The intrastriatal injection of an IL-1ß antibody (200 ng/2 µl) prevented the facilitation of GA-induced seizures by LPS. These data suggest that inflammatory processes during critical periods of development may decrease GA-induced seizure threshold.