Genetic variability of porcine parvovirus isolates revealed by analysis of partial sequences of the structural coding gene VP2

The 3'-terminal 853 nt (and the putative 283 aa) sequence of the VP2-encoding gene from 29 field strains of porcine parvovirus (PPV) were determined and compared both to each other and with other published sequences. Sequences were examined using maximum-parsimony and statistical analyses for nucleotide diversity and sequence variability. Among the nucleotide sequences of the PPV field strains, 26 polymorphic sites were encountered; 22 polymorphic sites were detected in the putative amino acid sequence. Mapping polymorphic sites of protein data onto the three-dimensional (3D) structure of PPV VP2 revealed that almost all substitutions were located on the external surface of the viral capsid. Mapping amino acid substitutions to the alignment between PPV VP2 sequences and the 3D structure of canine parvovirus (CPV) capsid, many PPV substitutions were observed to map to regions of recognized antigenicity and/or to contain phenotypically important residues for CPV and other parvoviruses. In spite of the high sequence similarity, genetic analysis has shown the existence of at least two virus lineages among the samples. In conclusion, these results highlight the need for close surveillance on PPV genetic drift, with an assessment of its potential ability to modify the antigenic make-up of the virus.

Desempenho de pontas de pulverização em Brachiaria brizantha cv. MG-4 para controle de ninfas de cigarrinhas das pastagens

O trabalho teve como objetivo estudar o desempenho de pontas de pulverização na deposição da calda inseticida para o controle de ninfas de cigarrinhas das pastagens em Brachiaria brizantha cv. MG-4. Doze tratamentos foram estudados em esquema fatorial 6x2, constituídos pelo contraste de seis pontas de pulverização e pressões de 196 e 392 kPa: TF-VP2 (336 L ha-1 e 467 L ha-1); AI11002-VS (184 L ha-1 e 200 L ha-1); XR11002-VS (200 L ha-1 e 280 L ha-1); TT11002-VP (200 L ha-1 e 280 L ha-1); TJ60-11002VS (208 L ha-1 e 280 L ha-1) e TX-VK4 (72 L ha-1 e 97 L ha-1). Para monitorar a deposição das caldas de pulverização, utilizaram-se os traçadores Azul Brilhante FD&C-1 (0,3% p/v) e Amarelo de Tartrasina FD&C-5 (0,6% p/v). Alvos artificiais, constituídos de lâminas de vidro, foram posicionados na base das plantas, próximos à superfície do solo, e os depósitos por unidade de área das soluções pulverizadas foram quantificados por espectrofotometria. As pontas TF-VP2, XR11002-VS e AI11002-VS, nas pressões de 196 e 392 kPa, proporcionam as maiores deposições da calda de pulverização na região das espumas das cigarrinhas das pastagens, apesar de apresentarem menor uniformidade na distribuição dos depósitos em relação a TX-VK4, XR110.02-VS e TJ110.02-VS. O aumento da pressão de 196 para 392 kPa promoveu aumento na deposição da calda de pulverização sobre a Brachiaria brizantha e na região onde se encontram as espumas das cigarrinhas para todos os tipos de pontas estudadas.

Replication of classical infectious bursal disease virus in the chicken embryo related cell line

Infectious bursal disease (IBD) is an acute, highly contagious viral disease. The diagnosis of IBD depends on time-consuming and costly procedures, like virus isolation on chick embryos and histopathological examination, A double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA), immunoperoxidase and reverse transcription polymerase chain reaction (RT-PCR) were applied in this study to detect classical IBD virus (IBDV) after three blind passages of the Lukert strain on chicken embryo related (CER) cell monolayer after different periods of infection: 6, 12, 24 and 48 h, Cytophatic effects were most evident 12 h post-infection (p.i.) but were observed at 6 h p.i. The maximum discrimination between IBDV-infected and uninfected cell suspensions obtained by the use of DAS-ELISA for virus detection corresponded to 0.597+/-0.02 and 0.010+/-0.01 after 12h p.i., respectively. The RT-PCR was performed using the set of primers A3.1 and A3.2 to amplify the VP2 region of the IBDV genome, This molecular technique demonstrated that from 6 h p.i., it was possible to detect the viral RNA. The results show that the CER cell line can be used for classical IBDV propagation, confirmed by the DAS-ELISA, immunoperoxidase and RT-PCR assay.

Frequência de anticorpos contra o vírus da língua azul em ovinos do estado do Ceará, Brasil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Substituição do uso de soro fetal bovino na manutenção do cultivo de células CER infectadas pelo vírus da doença infecciosa da bursa de Fabrícius

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Freqüência do parvovírus suíno em fetos mumificados, abortados e natimortos detectado através da reação em cadeia pela polimerase (PCR)

Porcine parvovirus (PPV) is associated with reproductive failure and it has been found worldwide, including Brazil. Many diagnostic procedures are used for its detection, for example, immunofluorescence, HA, HI and PCR and this is an important technique because it is very specific and sensitive. In this work, the presence of PPV in fetuses from swine farms with reproductive problems was detected by PCR. All of 170 samples from aborted fetuses, mummies or stillborns were sampled by PCR with primers designed to VP2 region of PPV and c-myc (endogenous control). Only 142 samples (83,53%) were positive for c-myc and among them six samples (4,22%) were positive for PPV which were tested in HA. In this test, erythrocytes suspension 1% was used and three samples (50%) agglutinated but they presented low titer (4). For this work, porcine parvovirus was detected in the samples analyzed by PCR and HA. It is important to emphasize the use of endogenous control when material with elevated degree of autolysis is examined