Анти-адгезивная стратегия в разработке комплексных противодифтерийных вакцин как перспективная мера снижения циркуляции Corynebacterium diphtheriae среди населения

A study is devoted to the development of new bacterial diphtheria vaccines aimed at prevention adhesion of microbial cells to host’s pharyngeal epithelial cells and thus limit colonization of mucous membranes, bacteria carrier state formation and promoted to C. diphtheriaе circulation decrease among the population. As a base of the candidate-vaccine we suggest the diphtheria bacterial antigenic preparation obtained by use of electro-magnetic radiation of ultrahigh frequency from a culture of C. diphtheriaе, var. gravis, tox +.Исследование посвящено разработке новых бактериальных дифтерийных вакцин, направленных на предупреждение колонизации слизистых оболочек, формирование бактерионосительства и способствующих снижению циркуляции C. diphtheriaе среди населения. В качестве основы кандидат-вакцины предлагается дифтерийный бактериальный антигенный препарат, полученный с помощью электро-магнитного излучения сверхвысокой частоты из культуры C. diphtheriaе, var. gravis, tox +

Theoretical confirmation of prospectivity of application of metabolitic complexes of lactobacilli and saccharomycetes in the fight antibiotic resistance of bacteria

Introduction. The global problem is the increase in the number of infectious diseases caused by antibiotic–resistant pathogens. Measures to control these microorganisms should be aimed at preventing the formation of antibiotic–resistant populations of microorganisms and at inhibiting already established resistant populations. The aim of the work is to substantiate the perspective of using metabolic complexes of lactobacilli and saccharomycetes to control antibiotic resistance of bacteria. Material & methods. Cellular structures of lactobacilli and saccharomycetes (L / S) were received by irradiation with low–frequency ultrasonic waves (generator G3–109) of suspensions of Lactobacillus rhamnosus GG (from symbiotic PREEMA®, Schonen, Switzerland) and Saccharomyces boulardii (from probiotic drug BULARDI®, Schonen, Switzerland). The metabolites of L. rhamnosus GG (ML) and S. boulardii (MS) were received in their own cellular structures. The combination of lactobacilli with saccharomycetes (MLS) and metabolites of saccharomycetes (LS) in the cellular structures of lactobacilli. Suspensions of microorganisms (resistant to antibiotics) gram–negative Pseudomonas aeruginosa PR, Acinetobacter baumannii PR, Klebsiella pneumoniae PR, Lelliottia amnigena (Enterobacter amnigenus) PR, gram–positive Staphylococcus aureus PR, Staphylococcus haemoliticus PR, Enterococcus faecalis PR, Corynebacterium xerosis PR, the cultures Corynebacterium spp. tox +, Streptococcus viridans, Streptococcus pneumoniaе with an optical density of 5,0 units on the McFarland scale (Densi–La–Meter (PLIVA–Lachema Diagnostika, (Czech Republic)) was added to L / S / ML / MLS / MS / LS (experimental samples) or to 0.9% sodium chloride solution (control samples) in a ratio of 1: 1. All samples were incubated for 1 hour at a temperature of + 35 ± 1 ° C, then the optical density was adjusted to 0.5 McFarland. Sowing was carried out on Mueller–Hinton medium. After disks with antibiotics (imipenem, vancomycin, cefotaxime, gentamicin, erythromycin, ciprofloxacin, amicil, chloramphenicol, ampicillin, ceftazidime , ceftriaxone, tetracycline, levofloxacin, amoxiclav, azithromycin), incubated (35 ± 1 ° C, 24 hours), measured the zones of growth retardation of microorganisms around the discs with antibiotics. Results & discussion. Potentiation of antimicrobial activity in the combined use of experimental samples with antibacterial drugs occurred in 88% of combinations with ML, 83 % – with MLS, 85 % – with MS, 73 % – with LS. Without increase in activity was substances with gentamicin, amicil, ampicillin, ceftazidime  were administered agains A. baumannii PR and with levofloxacin against S. aureus. Samples of ML over L (P = 0,005) and MLS over L (P = 0,008) had the advantage of a general increase in the diameters of the zones of growth inhibition of all tested pathogens. These results indicate a statistically significantly greater inhibition of growth of selected strains when combining antibiotics with metabolic complexes than with cellular structures. Excellent enhancement was observed when combining different antibacterial drugs with ML (on 5,5 ± 0,7, P <0,05), MLS (on 4,95 ± 0,6, P = 0,01) and MS (on 3,96±0,6, Р=0,001) relative to control. More inhibition of growth was observed of antibiotics with MLS than with MS (P = 0,02). A difference between the efficacies of the metabolic complexes ML and MLS was not found (P = 0,09). The presence of a large number of combinations of metabolic complex – antibiotic with the ability to therapeutically significant indicators to increase the antibacterial activity testifies the effectiveness of the combined use of metabolites L. rhamnosus GG and S. boulardii with different drugs. Conclusion. Theoretically confirmed of perspectivity of application of metabolic complexes of lactobacilli and saccharomycetes in the fight against antibiotic resistance of bacteria. Synergistic combinations of lactobacilli and saccharomycetes with antibiotics have been established. A therapeutically significant increase in their combined antimicrobial activity has been proven. This efficacy for different antibiotic–resistant strains indicates the perspectives of using metabolic complexes of lactobacilli and saccharomycetes to develop multifunctional antimicrobials preparations with consequence the possibility of inhibiting antibiotic resistance to already formed bacterial populations and at preventing the formation of antibiotic–resistant populations of microorganisms.DOI: 10.5281/zenodo.438221

Anti-adhesive Strategy in Development of Combined Antidiphtheritic Vaccine as a Perspective Preventive Measure of Corynebacteria diphtheriae Circulation Among the Population

A study is devoted to the development of new bacterial diphtheria vaccines aimed at prevention adhesion of microbial cells to host’s pharyngeal epithelial cells and thus limit colonization of mucous membranes, bacteria carrier state formation and promoted to C. diphtheriaе circulation decrease among the population. As a base of the candidate-vaccine we suggest the diphtheria bacterial antigenic preparation obtained by use of electro-magnetic radiation of ultrahigh frequency from a culture of C. diphtheriaе, var. gravis, tox +

METHODS OF CONTROL DIPHTHERIA VACCINE SAFETY

Vaccination success depends not only on the timely coverage of threatened contingents, but also on the quality of vaccines. Every day, the requirements for security guarantees vaccines and their use guarantees of security increases. For the fast, reliable and independent scientific assessment of vaccine safety issues, WHO in 1999 created the Global Advisory Committee on Vaccine Safety. To enhance the capacity of pharmaceutical supervision in relation to vaccines in 2012 it was developed the Global Vaccine Safety Initiative. The main directions of the Global Vaccine Safety programs are considered in this review. It’s noted more strict requirements of Ukrainian pharmaceutical industry to produce public immunization drugs regulated Supplements to the State Pharmacopoeia of Ukraine, in comparison with other countries. This review considered diphtheria vaccine safety monitoring in the process of production according to the recommendations of the World Health Organization (WHO), described a subcutaneous method for determining the specific toxicity of the combined purified toxoid, characterized an intracutaneous method of determining of the presence of diphtheria toxin in each sample of the combined purified toxoid, that additionally used by some manufacturers. The definition of diphtheria toxin in dilutions of purified toxoid is presented. This review considered diphtheria vaccine safety monitoring in the process of production according to the recommendations of the World Health Organization (WHO), described a subcutaneous method for determining the specific toxicity of the combined purified toxoid, characterized an intracutaneous method of determining of the presence of diphtheria toxin in each sample of the combined purified toxoid, that additionally used by some manufacturers. The definition of diphtheria toxin in dilutions of purified toxoid is presented. As methods for determination of diphtheria toxin must be able to detect even a small amount of toxin it's examined the WHO's proposal to use an intradermal method on guinea pigs and tests on cell cultures. Attention is drawn to the fact that the determination of specific toxicity in cell culture can be carried out at presence of the approval of this method of a national control authority and sensitivity rates no less than in experiments on guinea pigs. The determining of specific toxicity of ready vaccine by subcutaneous method is described. The publication gave a test for elevated toxicity of the final product by intraperitoneal infection of mice and guinea pigs. It’s cited the WHO recommendations aimed at removing the possibility of recovery of the refined toxin toxicity. Checking vaccines toxicity, pyrogenicity, sterility, allergenicity, teratogenicity, mutagenicity and immunogenicity mainly performed on laboratory animals. The review examined the unreliability of animal experiments and the need to find alternative methods for determining the toxicity without their use particularly in light of the “3R”concept. Methods for determining diphtherial toxin using cell cultures is considered, namely, colony overlay test (COT), tests using a monolayer of HeLa cell culture, a culture of Vero cells (kidney cells of african green monkeys ), a culture of CHO cells (cells of Chinese hamster ovary), which are based on the toxin cytopathic effect on sensitive cell culture. Their advantages and disadvantages are listed. An alternative method for the quantitative detection of C. diphtheriae toxin using the polystyrene plate coated with monoclonal antibody to the part of the diphtheria toxin which defines its binding to the cell, is described. It’s regarded the cytotoxic effect of diphtheria toxin on cells of the immune system of mice and guinea pigs: splenocytes, adhesive phagocytes i B- lymphocytes

Анти-адгезивная стратегия в разработке комплексных противодифтерийных вакцин как перспективная мера снижения циркуляции Corynebacterium diphtheriae среди населения

A study is devoted to the development of new bacterial diphtheria vaccines aimed at prevention adhesion of microbial cells to host’s pharyngeal epithelial cells and thus limit colonization of mucous membranes, bacteria carrier state formation and promoted to C. diphtheriaе circulation decrease among the population. As a base of the candidate-vaccine we suggest the diphtheria bacterial antigenic preparation obtained by use of electro-magnetic radiation of ultrahigh frequency from a culture of C. diphtheriaе, var. gravis, tox +.Исследование посвящено разработке новых бактериальных дифтерийных вакцин, направленных на предупреждение колонизации слизистых оболочек, формирование бактерионосительства и способствующих снижению циркуляции C. diphtheriaе среди населения. В качестве основы кандидат-вакцины предлагается дифтерийный бактериальный антигенный препарат, полученный с помощью электро-магнитного излучения сверхвысокой частоты из культуры C. diphtheriaе, var. gravis, tox +.</p