The use of spa and phage typing for characterization of a MRSA population in a Belgian hospital: comparison between 2002 and 2007.

UNLABELLED: TARGET OF THE STUDY: Strain typing of pathogens is essential to pinpoint the sources and routes of transmission and to forecast future trends. In a general hospital, we studied possible changes in the MRSA population. PATIENTS AND METHODS: MRSA isolates received from a Belgian general hospital, during 2002 (n=150) and the second half of 2007 (n=105), were compared by phage and spa typing. RESULTS: In 2002, [J]* phage types characterized 45% of the MRSA isolates, 13% belonged to the [O]* phage types, 12% to a local phage type 29/42E/54/D11* and 28% were not assigned to a defined group. Thirteen different spa types were found among the isolates: 39% belonged to t038, 27% to t121, 14% to t041, 5% to t740, and 4% to t002 and t024 each. Two spa types were found respectively in two and three isolates, five were unique. In 2007, 35% belonged to [J]*, 23% to [O]* and 39% could not be put in a defined group. Eighteen different spa types were found: 30% belonged to t740, 29% to t121, 13% to t038 and 10% to t002. Three spa types were represented in two isolates, eleven were unique. The t041 spa type was specific for the 29/42E/54/D11* and the majority of the t121 isolates were related to [J]*. CONCLUSION: [J]* remained the dominant phage types group but decreased whereas [O]*, the second phage types group, increased. As to the spa types, t740 became dominant while t121 remained second. Phage and spa typing point to some quantitative changes among the Belgian MRSA population.

Transformation of Brassica napus and Brassica oleracea Using Agrobacterium tumefaciens and the Expression of the bar and neo Genes in the Transgenic Plants.

An efficient and largely genotype-independent transformation method for Brassica napus and Brassica oleracea was established based on neo or bar as selectable marker genes. Hypocotyl explants of Brassica napus and Brassica oleracea cultivars were infected with Agrobacterium strains containing chimeric neo and bar genes. The use of AgNO(3) was a prerequisite for efficient shoot regeneration under selective conditions. Vitrification was avoided by decreasing the water potential of the medium, by decreasing the relative humidity in the tissue culture vessel, and by lowering the cytokinin concentration. In this way, rooted transformed shoots were obtained with a 30% efficiency in 9 to 12 weeks. Southern blottings and genetic analysis of S1-progeny showed that the transformants contained on average between one and three copies of the chimeric genes. A wide range of expression levels of the chimeric genes was observed among independent transformants. Up to 25% of the transformants showed no detectable phosphinotricin acetyltransferase or neomycin phosphotransferase II enzyme activities although Southern blottings demonstrated that these plants were indeed transformed.

Fusion of plant protoplasts: a study using auxotrophic mutants of Nicotiana plumbaginifolia, Viviani.

Protoplast fusion studies between various auxotrophic mutants of Nicotiana plumbaginifolia were performed to optimize conditions for PEG-mediated fusion and to identify factors influencing the plant protoplast fusion process. Numerous parameters in the isolation, culture, and fusion of protoplasts were tested, and established fusion protocols were compared. Fusion rates, calculated on the basis of colony growth on selection medium (genetic complementation), ranged from 10(-4) to 10(-2). Conditions that allow rapid and reproducible fusions at the highest rates were established. Particular emphasis was given to fusion of mesophyll-derived protoplasts, for which the ability to regenerate fertile plants from fusion products was shown to be particularly high. Preliminary experiments using electric-field mediated fusion suggest that electrofusion may offer significant advantages over the traditional chemical fusion.