A new analytical method based on capillary zone electrophoresis-tandem mass spectrometry (CZE-MS/MS) is proposed and validated for the identification and simultaneous quantification of eight quinolones for veterinary use in bovine raw milk. The studied quinolones include danofloxacin, sarafloxacin, ciprofloxacin, marbofloxacin, enrofloxacin, difloxacin, oxolinic acid, and flumequine, whose contents are regulated by the EU Council Regulation no. 2377/90 in animal edible tissues. Different parameters (i.e., separation buffer composition and electrospray conditions) were optimized in order to obtain both an adequate CE separation and a high sensitivity, using experimental design methodology to consider the interactions among the studied variables. MS/MS experiments using an ion trap as analyzer operating in the multiple reaction monitoring mode were carried out to achieve the minimum number of identification points according to the 2002/657/EC European Decision. For the quantification in bovine raw milk samples, a two-step solid-phase extraction procedure was developed using Oasis MAX and HLB cartridges without protein precipitation. Satisfactory results were obtained in terms of linearity (r2 between 0.989 and 0.992) and precision (RSD below 18%). The limits of detection and quantification (below 6 and 24 ppb, respectively) were in all cases lower than the maximum residues limits tolerated for these compounds in milk, the recoveries ranging from 81 to 110%, indicating the potential of the CZE-MS/MS for the analysis of regulated quinolone antibiotics in the food quality and safety control areas.

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The polytene chromosomes of Rhynchosciara americana and R. hollaenderi, a pair of sibling species in the americana-like group of Rhynchosciara, were compared using a number of techniques, including in situ hybridization. With classical cytological techniques, the only differences observed were in the morphology of centromeric and telomeric heterochromatin, in the size of a DNA and RNA puff, and in the presence of an inversion polymorphism in R. hollaenderi. However, after in situ hybridization with rDNA and poly-r(A) probes, differences between the two species appeared at a number of sites. Differences in poly-r(A) sites were especially informative in establishing phylogenetic relationships between these two species and a third species currently being examined from this group. Chromosomal evolution between these species appears to have occurred mainly through differential amplification and transposition of repetitive sequence DNA, of which dA:dT tracts are an important component. The R. hollaenderi karyotype is tentatively considered more ancestral than that of R. americana because it has features present in the third Rhynchosciara species. Explanations for the monomorphisms observed in Rhynchosciara species and mechanisms of speciation in the group are considered within the context of the species' complex behavior.

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