Highly synergistic antimicrobial activity of magainin 2 and PGLa peptides is rooted in the formation of supramolecular complexes with lipids

Magainin 2 and PGLa are cationic, amphipathic antimicrobial peptides which when added as equimolar mixture exhibit a pronounced synergism in both their antibacterial and pore-forming activities. Here we show for the first time that the peptides assemble into defined supramolecular structures along the membrane interface. The resulting mesophases are quantitatively described by state-of-the art fluorescence self-quenching and correlation spectroscopies. Notably, the synergistic behavior of magainin 2 and PGLa correlates with the formation of hetero-domains and an order-of-magnitude increased membrane affinity of both peptides. Enhanced membrane association of the peptide mixture is only observed in the presence of phophatidylethanolamines but not of phosphatidylcholines, lipids that dominate bacterial and eukaryotic membranes, respectively. Thereby the increased membrane-affinity of the peptide mixtures not only explains their synergistic antimicrobial activity, but at the same time provides a new concept to increase the therapeutic window of combinatorial drugs

Detection of Mycoplasma agalactiae by Polymerase Chain Reaction in Jordanian Sheep and Goat Herds

The aim of the study was to ascertain whether sheep and goats from selected Jordanian herds were infected with Mycoplasma agalactiae, the most common aetiological agent of contagious agalactia of sheep and goats. All examined animals showed clinical signs of disease at the time of sample collection. The group included 35 animals, 15 sheep and 20 goats. For microbiological examination, a total of 107 swabs were taken from conjunctival, nasal, vaginal or preputial mucosae and from the external auditory canal. Identification of the species isolated was carried out by a polymerase chain reaction. Of the 35 animals, 21 (4 sheep and 17 goats) tested positive for Mycoplasma agalactiae. These results confirmed our assumption that this mycoplasma species is present in Jordanian herds and, for the first time, provided evidence that contagious agalactia of sheep and goats occurs in Jordan

Light-Induced Nanosecond Relaxation Dynamics of Rhenium-Labeled Pseudomonas aeruginosa Azurins.

Time-resolved phosphorescence spectra of Re(CO)3(dmp)+ and Re(CO)3(phen)+ chromophores (dmp = 4,7-dimethyl-1,10-phenanthroline, phen = 1,10-phenanthroline) bound to surface histidines (H83, H124, and H126) of Pseudomonas aeruginosa azurin mutants exhibit dynamic band maxima shifts to lower wavenumbers following 3-exponential kinetics with 1-5 and 20-100 ns major phases and a 1.1-2.5 μs minor (5-16%) phase. Observation of slow relaxation components was made possible by using an organometallic Re chromophore as a probe whose long phosphorescence lifetime extends the observation window up to ∼3 μs. Integrated emission-band areas also decay with 2- or 3-exponential kinetics; the faster decay phase(s) is relaxation-related, whereas the slowest one [360-680 ns (dmp); 90-140 ns (phen)] arises mainly from population decay. As a result of shifting bands, the emission intensity decay kinetics depend on the detection wavelength. Detailed kinetics analyses and comparisons with band-shift dynamics are needed to disentangle relaxation and population decay kinetics if they occur on comparable timescales. The dynamic phosphorescence Stokes shift in Re-azurins is caused by relaxation motions of the solvent, the protein, and solvated amino acid side chains at the Re binding site in response to chromophore electronic excitation. Comparing relaxation and decay kinetics of Re(dmp)124K122Cu II and Re(dmp)124W122Cu II suggests that electron transfer (ET) and relaxation motions in the W122 mutant are coupled. It follows that nanosecond and faster photo-induced ET steps in azurins (and likely other redox proteins) occur from unrelaxed systems; importantly, these reactions can be driven (or hindered) by structural and solvational dynamics

Resistance to Antibiotics in Strains of Staphylococcus spp., Enterococcus spp. and Escherichia coli Isolated from Rectal Swabs of Pigs

The study aimed at determining the level of resistance of selected bacterial species (Staphylococcus spp., Enterococcus spp., Escherichia coli) isolated from rectal swabs of pigs to antimicrobial agents. The tested strains were isolated from piglets aged 7 to 30 days. Bacterial species were identified by standard microbiological techniques and susceptibility to antibiotics was determined quantitatively by the standard microdilution method. Resistance of the Staphylococcus aureus strain to oxacillin was confirmed by detection of the mecA gene and PBP2a. A total of 115 Staphylococcus spp. isolates were collected. In the case of Staphylococcus aureus, the methicillin-resistant strain (MRSA) was identified. Moreover, higher frequency of coagulase-negative staphylococci with minimum inhibitory concentration of oxacillin ≥ 0.5 mg/l was noticed. Inducible resistance to clindamycin in the Staphylococcus hominis strain was also detected. The strains of Enterococcus spp. (61 isolates) exhibited high resistance to tetracycline (98.5%), erythromycin (86.8%) and chloramphenicol (54.4%). Vancomycin-resistant enterococci were not isolated. In the case of Escherichia coli strains (111 isolates), higher frequency of resistant strains to tetracycline (81.1%) and ampicillin (62.2%) was documented. Resistance to fluoroquinolones and production of broad-spectrum β-lactamases was not noticed. The presented study may be considered as a pilot project assessing the prevalence of resistant bacteria in piglets kept on a single farm. It demonstrated the presence of resistant strains of Staphylococcus spp., including one MRSA strain, Enterococcus spp. and Escherichia coli. These strains may be present as a result of postnatal colonization with both bacterial microflora of dams and environmental microflora

COMPARISON OF METHODS TO MAP SELECTED TRAFFIC MARKINGS ON FIRST CLASS ROADS IN THE CZECH REPUBLIC

The article presents conclusions of a comprehensive analysis of pilot data collection using four mapping methods. To validate mapping methods and procedures, we selected three ten-kilometer sections of the first class roads with different geomorphological, vegetative and transportation properties. All sections were measured by aerial photogrammetry using GSD = 4 cm, mobile laser scanning equipment linked with cameras, by geodetic surveying methods, and one section was also measured by UAV. The tested methods mapped selected features of vertical and horizontal traffic markings on the first class roads. The traffic marking measuring sets were analyzed from the perspectives of personnel, time, data, costs, and technological and organizational aspects. All the mapping methods were verified as mentioned above starting from work preparation phase, its terrain realization, captured data processing and detailed analysis, concluding with stating the advantages and disadvantages for each mapping method. One of the analysis outputs was proposals to change and refine road administrator’s regulations. The mapping methods were compared with geodetic measurements. Analyses were also carried out in the context of creating digital data in 3D for the realization of BIM (Building Information Modeling) digital data in connection with the concept of the European Parliament and Council Directive 2014/24 / EU on Public Procurement, and Czech Government Decree 682 on the Concept of Implementation of the BIM Method in the Czech Republic of 25 September 2017 and Decree 958 of the Government of the Czech Republic of 2 November 2016 on the importance of BIM for the construction engineering and proposal of further steps to introduce it in the Czech Republic