pep27 and lytA in Vancomycin-Tolerant Pneumococci

Vancomycin therapy failure due to the emergence of tolerance in pneumococci is increasing. The molecular mechanism of tolerance is not clear, but lytA and pep27 are known to be involved. Our aim was to evaluate the expression of both genes in vancomycin-tolerant Streptococcus pneumoniae (VTSP) strains. Eleven VTSP strains from a total of 309 clinical isolates of S. pneumoniae from 1997 to 2006 were classified according to the criteria of Liu and Tomasz. All VTSP strains were evaluated for susceptibility according to CLSI criteria, serotype by the Quellung test, and clonality by PFGE. The expressions of lytA and pep27 were analyzed in different growth phases by RT-PCR with and without vancomycin. Eighty-two percent of VTSP strains showed resistance to penicillin, and 100% were sensitive to vancomycin and cefotaxime. The most frequent serotypes of VTSP strains were 23F (4/11) and 6B (3/11). Clonal relationship was observed in only two strains. No significant changes were observed in pep27 expression in the three phases of growth in VTSP strains with and without vancomycin. Interestingly, pep27 expression in the stationary phase in the non-tolerant reference strain R6 was significantly higher. However, no significant differences in lytA expression were observed between VTSP and R6 strains during the phases of growth analyzed. The absence of changes in pep27 expression in VTSP strains in the stationary phase may be related to their ability to tolerate high antibiotic concentrations, and thus, they survive and remain in the host under the antibiotic selective pressure reflected in therapeutic failure.     

Vegetation patterns,regeneration rates and divergence in an old-field succession of the high tropical Andes

Vegetation restoration during old-field succession was studied in an alpine Andean ecosystem (paramo). 123 plots with different fallow times (1 to 12 years) and 8 plots under natural vegetation were sampled. The results indicate that secondary succession in the paramo, like in other extreme environments, can be interpreted as an autosuccession: there are mainly changes in species relative abundance and little floristic relay (i.e. species turnover). Only a few herbaceous species, mostly introduced (e.g. Rumex acetosella), act as strict pioneers and strongly dominate the early stages. Then, they undergo a progressive decline, while native forbs (e.g. Lupinus meridanus) and grasses (e.g. Vulpia myuros) have their peak abundance in intermediate stages. The characteristic paramo life forms, sclerophilous shrubs (e.g. Baccharis prunifolia, Hypericum laricifolium) and giant rosettes (e.g. Espeletia schultzii), appear very early and gradually increase in abundance during succession, becoming dominant in the late stages and showing a dual behaviour, both as ruderal and stress tolerant species. The 1st axis of a Detrended Correspondence Analysis arranges the sites according to their fallow time. The 2nd and 3rd axes, associated with diverging pathways of regeneration, are correlated with topographic factors and physio-chemical soil characteristics. Hence, structural divergence between plots increases along succession as community composition starts to reflect the conditions of each site. We found evidence of a constant rate of succession during the first 12 years, contradicting the generally accepted hypothesis in the succession literature of a continous slow down up to the climax. Regeneration of vegetation physiognomy is relatively fast, questioning the prevailing idea of slow restoration in alpine ecosystems. However, 12 years of fallow are insufficient to attain the species richness of the natural paramo. Under the current trend of fallow length reduction observed in traditional potato cultivation in the Andes, our results raise doubts about the conservationist value of this management strategy.     

Population genetics of the endangered South American freshwater turtle, Podocnemis unifilis, inferred from microsatellite DNA data

We studied the population genetics of Podocnemis unifilis turtles within and among basins in the Orinoco and Amazon drainages using microsatellites. We detected high levels of genetic diversity in all sampled localities. However, ‘M-ratio’ tests revealed a substantial recent population decline in ten localities, in accord with current widespread exploitation. Our results reveal a consistent pattern across multiple analyses, showing a clear subdivision between the populations inhabiting the Amazon and Orinoco drainages despite a direct connection via the Casiquiare corridor, and suggesting the existence of two biogeographically independent and widely divergent lineages. Genetic differentiation followed an isolation-by-distance model concordant with hypotheses about migration. It appears that migration occurs via the flooded forest in some drainages, and via river channels in those where geographic barriers preclude dispersal between basins or even among nearby tributaries of the same basin. These observations caution against making generalizations based on geographically restricted data, and indicate that geographically proximate populations may be demographically separate units requiring independent management.     

Neotropical frogs and mating songs: The evolution of advertisement calls in glassfrogs

Anurans emit advertisement calls with the purpose of attracting mates and repelling conspecific competitors. The evolution of call traits is expected to be associated with the evolution of anatomical and behavioural traits due to the physics of call emission and transmission. The evolution of vocalizations might imply trade‐offs with other energetically costly behaviours, such as parental care. Here, we investigated the association between body size, calling site, parental care and call properties (call duration, number of notes, peak frequency, frequency bandwidth and call structure) of the advertisement calls of glassfrogs (Centrolenidae)—a family of Neotropical, leaf‐dwelling anurans—using phylogenetic comparative methods. We also explored the tempo and mode of evolution of these traits and compared them with those of three morphological traits associated with body size, locomotion and feeding. We generated and compiled acoustic data for 72 glassfrog species (46% of total species richness), including representatives of all genera. We found that almost all acoustic traits have significant, but generally modest, phylogenetic signal. Peak frequency of calls is significantly associated with body size, whereas call structure is significantly associated with calling site and paternal care. Thus, the evolution of body size, calling site and paternal care could constrain call evolution. The estimated disparity of acoustic traits was larger than that of morphological traits and the peak in disparity of acoustic traits generally occurred later in the evolution of glassfrogs, indicating a historically recent outset of the acoustic divergence in this clade.     

Pineapple (Ananas comosus L. Merr) micropropagation in temporary immersion systems

A procedure for the mass propagation of pineapple plants (Ananas comosus L. Merr) using a temporary immersion technique is described. This procedure involved three distinct phases in the automated temporary immersion system: shooting, bud differentiation and elongation. To establish this protocol, we used in vitro shoots obtained from established liquid culture as starting materials. Three culture methods (solid, liquid and temporary immersion) were compared. Temporary immersion increased the multiplication rate and fresh and dry weight after 42 days. Conventional micropropagation (liquid medium) and temporary immersion were compared in combination with paclobutrazol. Paclobutrazol promoted the formation of compact bud clusters with limited leaf development. The highest multiplication rate (106) was found when ex-plants were cultured in shooting medium (MS+2.1 mg/l BA+0.3 mg/l NAA) supplemented with 1 mg/l PB for 7 weeks. A 10-l temporary immersion bioreactor was used to test two approaches during elongation stage: reduction of the shoot-formation period or decrease of the initial number of explants. The highest number of competent and uniform plants (191.8 plant/l) was achieved when shoots were cultured for 4 weeks in shooting medium supplemented with PB. Received: 4 February 1998 / Revision received: 22 June 1998 / Accepted: 14 August 1998     

Depolarization and potentiation of responses to acetycholine elicited by ATP on frog muscle

Buchtal and collaborators reported thirty years ago an excitatory action of low (10⁻³M) concentrations of ATP on frog muscle, as well as in increase in the sensitivity of the muscle to ACh. These effects have been re-investigated employing both intra-and extra-cellular recording and the technique of iontophoretic drug application. ATP at a concentration of 10⁻⁴M decreases the recorded resting potential by about 35%. The depolarizing action of ATP is more pronounced in the tibial end of the frog sartorius muscle than in the nerve free pelvic end. In addition ATP, added to the bath and electro-osmotically applied, increases the depolarizing action of ACh. This potentiating effect is particularly marked in denervated muscles.     

The physiology of ex vitro pineapple (<Emphasis Type="Italic">Ananas comosus</Emphasis> L. Merr. var MD-2) as CAM or C3 is regulated by the environmental conditions

Many plant species grown under in vitro controlled conditions can be used as models for the study of physiological processes. Adult pineapple can display CAM physiology while in vitro it functions as a C3 plant. Ex vitro Ananas comosus has plastic morphology and physiology, both easy to modify from C3 to CAM by changing the environmental conditions. The yield of survival for a rentable propagation protocol of pineapple is closely related with the C3/CAM shift and the associated physiological characteristics. In the present work, ex vitro pineapple plants were divided in two sets and subjected to C3 and CAM-inducing environmental conditions, determined by light intensity and relative humidity, respectively, 40 μmol m⁻² s⁻¹/85% and 260 μmol m⁻² s⁻¹/50%. The results demonstrated that the stress imposed by the environmental conditions switched pineapple plants from C3 to CAM behavior. Comparing to CAM induced, C3-induced pineapple plants showed substandard growth parameters and morphological leaf characteristics but a better rooting process and a higher ABA production, a phenotype closer to adult plants, which are expected to produce fruits in a normal production cycle. We conclude that the upholding of these characteristics is conditioned by low light intensity plus high relative humidity, especially during the first 8 weeks of ex vitro growth. It is expected that the better understanding of pineapple acclimatization will contribute to the design of a protocol to apply as a rentable tool in the pineapple agronomic industry.