Prevalence of spotted fever group rickettsiae in Ixodes ricinus (Acari: Ixodidae) in southern Germany.

Host-seeking Ixodes ricinus (L.) (Acari: Ixodidae) ticks were collected systematically, from May to September 2006, at selected sites in southern Germany, including a large city park in Munich. Polymerase chain reactions for amplification of genes of the rickettsial citrate synthase (gltA), the outer membrane proteins A and B (ompA and ompB), and the 16S rDNA were used to investigate 2,861 specimens (adults and nymphs). GltA sequences of spotted fever group rickettsiae were detected in 151 of all samples (5.3%; 95% CI = 4.3-6.2%). Sequencing revealed Rickettsia helvetica in 91.4% of the samples and R. monacensis in 8.6%. Amplification of ompA was not possible for R. helvetica, but in all except one of the R. monacensis. The results were analyzed statistically to test the effects of season, location, developmental stage, and gender of the tick on prevalence of Rickettsia spp. Although rickettsial DNA was detected in all investigated sites, sites in natural forest areas had significantly higher prevalences than sites in landscaped city parks. Adult female and male ticks had a similar prevalence and were significantly more often infected than nymphs. Monthly differences were not statistically significant. These results clearly show that R. helvetica is widespread throughout the study region and could result in a threat to public health in areas of high prevalence.

Rickettsia felis in fleas, Germany.

Among 310 fleas collected from dogs and cats in Germany, Rickettsia felis was detected in all specimens (34) of Archaeopsylla erinacei (hedgehog flea) and in 9% (24/226) of Ctenocephalides felis felis (cat flea). R. helvetica was detected in 1 Ceratophyllus gallinae (hen flea).

The human combinatorial antibody library HuCAL GOLD combines diversification of all six CDRs according to the natural immune system with a novel display method for efficient selection of high-affinity antibodies.

This article describes the generation of the Human Combinatorial Antibody Library HuCAL GOLD. HuCAL GOLD is a synthetic human Fab library based on the HuCAL concept with all six complementarity-determining regions (CDRs) diversified according to the sequence and length variability of naturally rearranged human antibodies. The human antibody repertoire was analyzed in-depth, and individual CDR libraries were designed and generated for each CDR and each antibody family. Trinucleotide mixtures were used to synthesize the CDR libraries in order to ensure a high quality within HuCAL GOLD, and a beta-lactamase selection system was employed to eliminate frame-shifted clones after successive cloning of the CDR libraries. With these methods, a large, high-quality library with more than 10 billion functional Fab fragments was achieved. By using CysDisplay, the antibody fragments are displayed on the tip of the phage via a disulfide bridge between the phage coat protein pIII and the heavy chain of the antibody fragment. Efficient elution of specific phages is possible by adding reducing agents. HuCAL GOLD was challenged with a variety of different antigens and proved to be a reliable source of high-affinity human antibodies with best affinities in the picomolar range, thus functioning as an excellent source of antibodies for research, diagnostic, and therapeutic applications. Furthermore, the data presented in this article demonstrate that CysDisplay is a robust and broadly applicable display technology even for high-throughput applications.