The mechanisms of complement activation in normal bovine serum and normal horse serum against Yersinia enterocolitica O:9 strains with different outer membrane proteins content.

Yersinia enterocolitica is a common zoonotic pathogen and facultative intracellular bacterium which can survive within blood cells. Cattle and horses are considered a reservoir of Y. enterocolitica which often causes several serious syndromes associated with yersiniosis such as abortions, premature births or infertility. The aim of our investigation was to determine the vitality of Y. enterocolitica O:9 strains (Ye9) in bovine and horse sera (NBS and NHrS) and explain the role of outer membrane proteins (OMPs) in serum resistance of these bacteria. Our previous studies demonstrated moderate human serum (NHS) resistance of the wild type Ye9 strain, whereas mutants lacking YadA, Ail or OmpC remained sensitive to the bactericidal activity of NHS. The present study showed that the wild type of Ye9 strain was resistant to the bactericidal activity of both NHrS and NBS, while Ye9 mutants lacking the YadA, Ail and OmpC proteins were sensitive to NHrS and NBS as well as to NHS. The mechanisms of complement activation against Ye9 strains lacking Ail and YadA were distinguished, i.e. activation of the classical/lectin pathways decisive in the bactericidal mechanism of complement activation of NBS, parallel activation of the classical/lectin and alternative pathways of NHrS. In this research the mechanism of independent activation of the classical/lectin or the alternative pathway of NBS and NHrS against Ye9 lacking OmpC porin was also established. The results indicate that serum resistance of Ye9 is multifactorial, in which extracellular structures, i.e. outer membrane proteins (OMPs) such as Ail, OmpC or YadA, play the main role.

Silver Nanoforms as a Therapeutic Agent for Killing Escherichia coli and Certain ESKAPE Pathogens.

The scope of this study included the preparation of silver nanoforms with high antimicrobial efficacy, low cost, and ease of application. The term 'silver nanoforms' refers to silver located on the amorphous or crystalline titanium dioxide (TiO2). Silver nanoforms may be used as an alternative to antibiotics in killing bacteria. Pure and silver-incorporated titanium (used as a carrier) was prepared using the sol-gel-modified method. Physical and chemical properties of the samples were described, and the antibacterial activity was indicated using the following strains of bacteria: Staphylococcus aureus, Klebsiella pneumoniae (ESKAPE pathogens), and Escherichia coli. The results have shown that the antibacterial activity of silver nanoforms with amorphous TiO2 is much better than that in the samples based on anatase (crystalline TiO2). The sensitivity of the tested bacteria to silver nanoforms depends on physical and chemical properties of the nanoforms and individual characteristics of the bacteria. For the first time, significant participation of amorphous TiO2 in antibacterial compounds has been described through this study.

New photosensitive nanometric graphite oxide composites as antimicrobial material with prolonged action.

A new conjugate material based on partially reduced graphite oxide (rGO), silver nanoparticles (Ag), and bis(lysinato)zirconium(IV) phthalocyanine complex (ZrPc) was obtained. Its optical properties (absorption and photoluminescence) after dispersion in solvents were examined. The antimicrobial properties were tested to determine the effect of the composite on the following bacterial strains: Staphylococcus aureus, Pseudomonas aeruginosa, and Escherichia coli, which are responsible for many infections and are one of the pathogens the most difficult to treatment. The results obtained for rGO-ZrPc-Ag composite were compared with the properties of GO, GO-ZrPc, and rGO-Ag structures. The influence of the near-infrared irradiation on the antimicrobial activity of ZrPc- and Ag-doped materials against bacteria was observed for very low concentration (0.32mg/mL) of GO-ZrPc to stop the growth of P. aeruginosa in comparison to the nonirradiated sample (41mg/mL). The usefulness of this material in therapy, such as wound infection treatment or endodontic treatment, as antibacterial agent with sustained action was discussed.

Selection and electrophoretic characterization of Salmonella enterica subsp. enterica biocide variants resistant to antibiotics.

The proposed research outlines a serious common concern of Salmonella resistance to antimicrobials following prolonged exposure to the disinfectants (biocides). These phenotypes of bacteria could potentially result in hard to treat infections. Typical for avian sources, biocide sensitive S. enterica subsp. enterica serovars: Typhimurium, Enteritidis, Virchow and Zanzibar and their isogenic biocide-tolerant variants were studied in order to investigate bacteriostatic effect of two commercially available biocide formulations: potassium peroxymonosulfate (P) and dodecylamine based structure (triamine, D). We found that cultivating of the bacteria in the medium supplemented with a blend containing P did not influence their antibiotic susceptibility pattern. In contrast, tolerance of bacteria to D compound resulted in resistance to co-trimoxazole, cefotaxime and ciprofloxacin of which two cefotaxime and ciprofloxacin are used commonly for the treatment of invasive Salmonella infections in humans. The dependency between OMP patterns and the level of Salmonella survival in media containing the biocides was observed merely in serovar Typhimurium. In conclusion, these results suggest that Salmonella strains challenged by prolonged treatment with the disinfectants become resistant to antibiotics, however it depends on Salmonella serovar and the chemical used. This paper also highlights the loop-mediated isothermal amplification (LAMP) as a technique that offers great benefits to microbiological detecting of Salmonella species in the samples.

Application of zwitterionic detergent to the solubilization of Klebsiella pneumoniae outer membrane proteins for two-dimensional gel electrophoresis.

Klebsiella pneumoniae is a frequent cause of nosocomial respiratory, urinary and gastrointestinal tract infections and septicemia with the multidrug-resistant K. pneumoniae being a major public health concern. Outer membrane proteins (OMPs) are important virulence factors responsible for the appropriate adaptation to the host environment. They constitute of the antigens being the first in contact with infected organism. However, K. pneumoniae strains are heavily capsulated and it is important to establish the OMPs isolation procedure prior to proteomics extensive studies. In this study we used Zwittergent Z 3-14® as a detergent to isolate the OMPs from K. pneumoniae cells and resolve them using two-dimensional electrophoresis (2-DE). As a result we identified 134 protein spots. The OMPs identified in this study are possible candidates for the development of a protein-based vaccine against K. pneumoniae infections.

The role of complement activity in the sensitivity of Salmonella O48 strains with sialic acid-containing lipopolysaccharides to the bactericidal action of normal bovine serum.

Sialic acids are important constituents of animal tissue glycoconjugates and are also present in the antigens of some bacterial strains. Capsular polysaccharides with sialic acid (NeuAc) have been extensively studied with regard to sensitivity to the bactericidal action of serum, whereas little is known in this regard about lipopolysaccharides (LPS) which contain NeuAc. Strains of Salmonella O48, able to infect animals and containing the same structures of LPS with NeuAc, were examined for their susceptibility to the bactericidal action of normal bovine serum (NBS). The strains showed varied sensitivity to the bactericidal action of NBS, which indicates that the expression of LPS containing NeuAc residues is not critical for the strains' resistance to the serum's activity. In this study the mechanisms of complement activation responsible for killing serum-sensitive Salmonella O48 rods by NBS were also established. Three such mechanisms were distinguished: activation of the classical/lectin pathways, important (decisive) in the bactericidal mechanism of complement activation, parallel activation of the classical/lectin and alternative pathways, and independent activation of the classical and lectin or the alternative pathway.

Killing of Gram-negative bacteria with normal human serum and normal bovine serum: use of lysozyme and complement proteins in the death of Salmonella strains O48.

Serum is an environment in which bacterial cells should not exist. The serum complement system provides innate defense against microbial infections. It consists of at least 35 proteins, mostly in pre-activated enzymatic forms. The activation of complement is achieved through three major pathways: the classical, alternative, and lectin. Lysozyme, widely present in body fluids, catalyzes the hydrolysis of beta 1,4 linkage between N-acetyloglucosamine and N-acetylmuramic acid in the bacterial cell wall and cooperates with the complement system in the bactericidal action of serum. In this study, ten strains of serotype O48 Salmonella, mainly associated with warm-blooded vertebrates and clinically important causing diarrhea in infants and children, were tested. The results demonstrated that the most efficient killing of Salmonella O48 occurred when all the components of normal bovine serum (NBS) and normal human serum (NHS) cooperated. To prove the role of lysozyme in the bactericidal activity of bovine and human serum, the method of serum adsorption onto bentonite (montmorillonite, MMT) was used. In order to investigate structural transitions accompanying the adsorption of serum components, we applied X-ray diffraction methods. The results of this investigation suggested that apart from lysozyme, other proteins (as, e.g., C3 protein or IgG immunoglobulin) were adsorbed on MMT particles. It was also shown that Ca(2+) cations can be adsorbed on bentonite. This may explain the different sensitivities of the serovars belonging to the same O48 Salmonella serotype to NBS and NHS devoid of lysozyme.

The mechanisms of activation of normal human serum complement by Escherichia coli strains with K1 surface antigen.

Ten E. coli K1 strains isolated from the urine of children with urinary tract infections were sensitive to the bactericidal action of normal human serum (NHS). The role of the particular mechanisms of complement activation was determined in the process of killing these strains, showing variable sensitivity to the bactericidal action of NHS; three mechanisms of activation of human complement were observed. Important role of alternative pathway activation in the bactericidal action of NHS against E. coli K1 strains independent of the classical and lectin pathways was not established.