Summary A live attenuated cereal‐based classical swine fever (CSF) vaccine for oral application, a form of vaccine that farmers can use themselves, demonstrated the improvement of CSF control in backyard production systems in endemic areas. Due to the dependency on very low storage temperature (−20°C) of the cereal‐based oral CSF vaccine, a lyophilized cereal‐based oral vaccine has been developed and tested. Although some studies showed total protection against a virulent virus strain, the production procedure is still considered complex. In this study, the lyophilized oral CSF virus, which is easy to produce and could be kept in the form of a bread‐based vaccine at 4°C for an extended time, was tested for the ability to induce a humoral immune response in pigs. Among the materials tested as a base for the CSF virus vaccine, plain sliced bread was considered the most appropriate base because of its absorbing ability and virus titre maintenance. Titres of bread‐based lyophilized CSF virus vaccine were stable at around 3.67 log10 TCID50 per ml for 7 months at 4°C. Pigs aged 5 and 8 weeks that orally received five bread‐based lyophilized CSF virus vaccine showed seroconversion of over 90% at 14 dpv. At 28 dpv, both age groups showed 100% seroconversion. In conclusion, the bread‐based lyophilized CSF virus vaccine can be an alternative choice for oral vaccination of pigs. Due to the simple process of production and the need for less virus titre, vaccine prices could be lowered. However, vaccine thermostability has to be improved to allow the vaccine delivery to be less dependent on functioning cool chains.

Dihydroflavonol 4-reductase (DFR) gene is a key gene of anthocyanins biosynthesis pathway, which represent an importance pathway for orchid flower. In this study, cloning and expression analysis of DFR gene in Ascocenda spp. were carried out. Nucleotide analysis revealed that the Ascocenda DFR gene was 1,056 bp in length, and encoded a protein of 351 amino acid residues. A typical translation initiation codon (ATG) and translation termination codon (TGA), the most frequently found codon in plant were identified, indicating a full-length coding sequence of the DFR gene. The calculated molecular mass of the deduced polypeptide was 39.8 kDa and the predicted isoelectric point was 5.58. Homology analysis revealed that the amino acid sequence of the Ascocenda DFR gene product was 80 to 87% identity to amino acid sequences of DFR gene products of other orchids such as Bromheadia, Dendrobium, Cymbidium and Oncidium. It also showed a high degree of identity to the DFR gene products of other flowers such as Lilium, Tilipa, Allium, Gentiana and Chrysanthenum. Southern blot analysis indicate that DFR is presented as a single copy in the Ascocenda spp. genome. The AscoDFR gene was highly expressed in the flower stages 2 and 3 of development as well as in the sepal and petal of the orchid flower.