Host feeding patterns of Culicoides species (Diptera: Ceratopogonidae) within the Picos de Europa National Park in northern Spain.

Blood meal identification can provide information about the natural host-feeding patterns or preferences of Culicoides species. Such information could indirectly provide data indicating which reservoirs are significant in associated vector-borne diseases. We positively identified the host species through DNA sequencing of the cytochrome b gene in 144 of the 170 (84.7%) blood meal specimens tested. In the remaining samples, identification of the blood-meal source was unsuccessful, possibly due to the post-ingestion time prior to sampling or the availability of the species-specific cytochrome b gene sequences in the database. The majority of identified blood meals were derived from mammalian blood (95.8%), and only six contained chicken blood. We identified five species as mammalian hosts for Culicoides spp.: sheep (87.7%), human (6.5%), cattle (3.7%) and Savi's Pine Vole (Micrototus savii) (2.1%). The results suggested that large mammals, specifically ruminants, were most frequently fed upon by biting midges (Culicoides spp.), but evidence of opportunistic feeding behaviour was also found. Host feeding behaviour of Culicoides species may also be influenced by the relative abundance of a particular host species in the area being studied. In this sense, Savi's Pine Vole, a wild species, was found to be a locally relevant host and a putative reservoir for viruses transmitted by species of biting midges belonging to the Culicoides genus. Finally, feeding on multiple potential host species was observed. One midge acquired blood meals from human and chicken hosts, while four other midges fed on two different sheep.

Glyceraldehyde-3-phosphate dehydrogenase-encoding gene as a useful taxonomic tool for Staphylococcus spp.

The gap gene of Staphylococcus aureus, encoding glyceraldehyde-3-phosphate dehydrogenase, was used as a target to amplify a 933-bp DNA fragment by PCR with a pair of primers 26 and 25 nucleotides in length. PCR products, detected by agarose gel electrophoresis, were also amplified from 12 Staphylococcus spp. analyzed previously. Hybridization with an internal 279-bp DNA fragment probe was positive in all PCR-positive samples. No PCR products were amplified when other gram-positive and gram-negative bacterial genera were analyzed using the same pair of primers. AluI digestion of PCR-generated products gave 12 different restriction fragment length polymorphism (RFLP) patterns, one for each species analyzed. However, we could detect two intraspecies RFLP patterns in Staphylococcus epidermidis, Staphylococcus hominis, and Staphylococcus simulans which were different from the other species. An identical RFLP pattern was observed for 112 S. aureus isolates from humans, cows, and sheep. The sensitivity of the PCR assays was very high, with a detection limit for S. aureus cells of 20 CFU when cells were suspended in saline. PCR amplification of the gap gene has the potential for rapid identification of at least 12 species belonging to the genus Staphylococcus, as it is highly specific.