Multiresistance in Pasteurella multocida is mediated by coexistence of small plasmids

In most gram-negative bacteria, acquired multiresistance is conferred by large plasmids compiling numerous antimicrobial resistance genes. Here, we show an evolutionary alternative strategy used by Pasteurella multocida to become resistant to multiple clinically relevant antibiotics. Thirteen beta-lactam-resistant clinical isolates, concomitantly resistant to tetracyclines and/or streptomycin as well as to sulfonamides, were studied. Pulsed-field gel electrophoresis analysis revealed different profiles among the isolates, showing that clonal dissemination was not the sole event responsible for the spread of multiresistance. Each P. multocida strain carried two or three small plasmids between 4 and 6 kb in size. A direct association between resistance profile and plasmid content was found. Complete nucleotide sequencing of all plasmids revealed seven different replicons, six of them belonging to the ColE1 superfamily. All plasmids carried one, or a maximum of two, antimicrobial resistance determinants. Plasmids pB1000 and pB1002 bore bla(ROB-1), pB1001 carried tet(B), pB1003 and pB1005 carried sul2 and strA, pB1006 harbored tet(O), and p9956 bore the tet(H) gene. All plasmids except pB1002 and pB1006 were successfully transformed into Escherichia coli. pB1000, also involved in beta-lactam resistance in Haemophilus parasuis (A. San Millan et al., Antimicrob. Agents Chemother. 51:2260-2264, 2007), was mobilized in E. coli using the conjugation machinery of an IncP plasmid. Stability experiments proved that pB1000 was stable in P. multocida but highly unstable in E. coli. In conclusion, bla(ROB-1) is responsible for beta-lactam resistance in P. multocida in Spain. Coexistence and the spread of small plasmids are used by P. multocida to become multiresistant.

Quantifying the contamination by old main-sequence stars in young moving groups: the case of the Local Association

Context. The associations and moving groups of young stars are excellent laboratories for investigating stellar formation in the solar neighborhood. Previous results have confirmed that a non-negligible fraction of old main-sequence stars is present in the lists of possible members of young stellar kinematic groups. A detailed study of the properties of these samples is needed to separate the young stars from old main-sequence stars with similar space motion, and identify the origin of these structures. Aims. Our intention is to characterize members of the young moving groups, determine their age distribution, and quantify the contamination by old main-sequence stars, in particular, for the Local Association. Methods. We used stars possible members of the young (~10-650 Myr) moving groups from the literature. To determine the age of the stars, we used several suitable age indicators for young main sequence stars, i.e., X-ray fluxes from the Rosat All-sky Survey database, photometric data from the Tycho-2, Hipparcos, and 2MASS database. We also used spectroscopic data, in particular the equivalent width of the lithium line Li I λ6707.8 Å and H_α, to constrain the range of ages of the stars. Results. By combining photometric and spectroscopic data, we were able to separate the young stars (10-650 Myr) from the old (> 1 Gyr) field ones. We found, in particular, that the Local Association is contaminated by old field stars at the level of ~30%. This value must be considered as the contamination for our particular sample, and not of the entire Local Association. For other young moving groups, it is more difficult to estimate the fraction of old stars among possible members. However, the level of X-ray emission can, at least, help to separate two age populations: stars with <200 Myr and stars older than this. Conclusions. Among the candidate members of the classical moving groups, there is a non-negligible fraction of old field stars that should be taken into account when studying the stellar birthrate in the solar neighborhood. Our results are consistent with a scenario in which the moving groups contain both groups of young stars formed in a recent star-formation episode and old field stars with similar space motion. Only by combining X-ray and optical spectroscopic data is it possible to distinguish between these two age populations.

Evolution of specific antibodies and proviral DNA in milk of small ruminants infected by small ruminant lentivirus

The diagnosis of Small Ruminant Lentivirus (SRLV) is based on clinical signs, pathological lesions and laboratory testing. No standard reference test for the diagnosis of maedi visna has been validated up to the present, and it is puzzling that tests which detect antibodies against the virus and tests which detect the proviral genome may render opposite results. The aim of this study was to evaluate the presence in milk throughout a lactation period of specific antibodies by ELISA and of SRLV proviral DNA by a PCR of the highly conserved pol region. A six-month study was conducted with the milk of 28 ewes and 31 goats intensively reared. The percentage of animals with antibodies against SRLV increased throughout the study period. Seroprevalence in sheep was 28% at the beginning of the study and by the end it had increased up to 52.4%. In goats, initial seroprevalence of 5.6% increased to 16%. The percentage of PCR positive ewes was stable throughout the study period. Of the positive sheep, 21.4% were PCR-positive before antibodies could be detected and most of them became PCR-negative shortly after the first detection of antibodies. This might suggest that antibodies have a neutralizing effect. In addition, an equal percentage of sheep were always PCR-negative but either became ELISA-positive or was always ELISA-positive, which might support this hypothesis. On the other hand, the PCR results in goats did not follow any pattern and oscillated between 35.3% and 55.6% depending on the month. Most goats positive by PCR failed to develop antibodies in the 6 months tested. We may conclude that the infection and the antibody response to it follow a different trend in sheep and goats.