Vol 91, No 1 (2014)

Aim. Determination of penicillin resistance features in tularemia causative agents of the mediasiatica subspecies, stability evaluation of differences in strains of various taxa and development of a rapid method of F. tularensis intraspecies differentiation. Materials and methods. β-Lactamase activity was determined in 30 strains of Francisella genus bacteria by quantitative iodometric method. Results. All the strains regardless of subspecies membership were characterized by high resistance to β-lactam antibiotics, and bacteria of the mediasiatica subspecies in contrast to other francisella did not synthesize β-lactamase. Our attempts to induce β-lactamase activity in vitro and in vivo in strains of this subspecies did not succeed. A method of intraspecies differentiation of F. tularensis by nitorcefin disks is proposed based on the distinctive feature. Conclusion. A high level of F. tularensis subsp. mediasiatica resistance to β-lactams in vitro and their inefficiency during therapy of experimental tularemia due to a β-lactamase negative strain suggests that F. tularensis β-lactamase is not the leading factor in formation of native penicillin resistance of tularemia causative agent.

Aim. Study of Bordetella pertussis lipopolysaccharide (LPS) immunobiological properties in the acellular pertussis vaccine. Materials and methods. Experimental series of acellular pertussis vaccines (APV), lyophilized LPS were used. Antibody titers against LPS in mice sera were evaluated by using EIA with peroxidase conjugate of anti-species antibodies against mice IgG. LPS activity in B. pertussis antigen complex preparations was determined in quantitative chromogenic LAL-test by end point. APV protective activity was determined in mice test during intracerebral infection by B. pertussis strain No. 18323 virulent culture. APV safety was determined in the mice body weight change test. Results. The presence of LPS in APV was shown in immune electrophoresis with purified B. pertussis LPS preparation as a control. Formalin treatment changes immunochemical properties of APV LPS that lead to the shift of precipitation bands with pertussis agglutinating sera from the start zone into cathode. The quantity of LPS in pertussis culture supernatants was on average 49050+6774 endotoxin units per ml (EU/ml). In APV preparations the quantity of LPS was on average 906+90 EU/ml, i.e. decreased by more than 50 times. An increase of antibody titers against B. pertussis LPS in mice sera after the APV immunization was shown in EIA, which gives evidence of its presence in immunogenic form in the complex preparations. The preclinical studies carried out show protective activity and specific safety of the experimental APV series. Conclusion. Formalin-neutralized APV preparation is a complex of protein antigens in association with LPS. Formalin treatment results in modification of LPS molecule that retains antigenic properties but is significantly less toxic.

Aim. Production, study ofproperties and evaluation ofa possibility to use monoclonal antibodies against Pseudomonas aeruginosa exotoxin A (ETA) for the detection ofmolecules ofrecombinant exotoxin A and anatoxin. Materials and methods. Producer hybridomas for monoclonal antibodies against ETA were generated. Recombinant exotoxin A, its atoxic forms, P.aeruginosa anatoxin and P. aeruginosa PA-103 live culture were used for immunization of mice. Results. The experiment of fusion of malignant cell line with immune lymphocytes obtained from a mouse immunized with recombinant ETA turned out to be the most productive. Intensity of interaction of monoclonal antibodies with recombinant atoxic forms ofETA was evaluated in enzyme immunoassay. Protective (toxin-neutralizing) activity of antibodies in cell culture and the ability to detect exotoxin A in latex-agglutination reaction were studied. Antibodies of hybrid culture No. 21 are able to detect molecules of recombinant ETA and anatoxin in the latex-agglutination reaction. Conclusion. Use of the antibodies produced for testing toxigenicity of P. aeruginosa strains as well as detection of anatoxin in the technological process of production of prophylaxis preparations based on exotoxin A is expected.

Aim. Evaluation of photodynamic therapy (PDT) in vitro antiviral effect directly on the herpes simplex virus (HSV) as well as on virus infected cells. Materials and methods. HSV infected \fero cell culture; HSV-1 (VR-3 strain) and HSV-2 (MS strain) were used. Fotoditazin preparation (chlorin E6 derivative) was used as a photosensitizer. AFS physiotherapy apparatus was used for laser irradiation. Experiment variants: study of direct and mediated photodynamic effect on virus-containing fluid (HSV-1 and HSV-2) and on non-infected cell culture; evaluation of anti-herpetic effect of PDT and study of the effect on the newly produced HSV-1 progeny were carried out within the developed experiment scheme. Results. A scheme of experiment conduction for study of mediated photodynamic effect of HSV was developed. A significant reduction of virus titers for more than 2 orders of magnitude (100-1000 times) was detected. Optimal concentration of photosensitizer and laser irradiation dose that results in reduction of HSV-1 titers by 1000 and more times (1.5-2.5 orders of magnitude) and reduction of HSV-2 titers by 10 and more times (1.5 order of magnitude) were determined. Conclusion. The data obtained will help selection of adequate scheme of PDT conduction for patients with recurrent herpes virus infection including patients with recurrent miscarriage.

Aim. Study the influence of antibiotic therapy on the formation of main biotope microflora (nasopharynx, large intestine, urinary system) of the premature child organism weighing less than 1500 g at birth in hospital. Materials and methods. Bacteriological study of upper respiratory tract discharge, urine, large intestine contents in 58 premature children during admission and discharge from newborn and premature children pathology department was carried out. Factor analysis method was applied to construct factor models of antibiotics influence on main biotope micro-biocenosis formation. Results. Deficiency of obligatory normal flora members in all the 3 biotopes was noted in all the children during admission to the department. Colonization of all the biotopes by obligatory members was noted by discharge, however the parameters do not reach age norm. Antibiotics therapy is the main factor, in all the biotopes opportunistic and pathogenic microorganisms prevail. Conclusion. The formation of main biotope microbiocenoses does not occur and carriage of pathogenic and opportunistic microorganisms is noted in premature children receiving massive antibiotic therapy.

Experience of Olympic Games host-countries in procuring protection from a biological threat is examined. Based on analysis of measures directed at improvement of systems of epidemiologic control and bioterrorism counteraction at London Olympics (2012), recommendations are given on the main directions of organizational, prophylactic and anti-epidemic activities during mass events.

Contemporary experience of organization of atmospheric air monitoring for the presence of pathogenic biological agents (PBA) is generalized in the article. Proposals for the use of posts of atmospheric air control during mass events are presented. Issues of equipping the instrument base for air sampling and PBA detection are examined.

The review is dedicated to immunologic adjuvants - various natural and synthetics substances that are added to vaccines for stimulation of specific immune response, but they do not induce specific response themselves. Critically important is the selection of the correct adjuvants, for which mechanisms of effect on immune system are studied the most. The majority of these mechanisms as well as physical-chemical and biological features of modern adjuvants are analyzed in the review. The problem of safety of adjuvants, types of immune response induced by adjuvants of various nature, excipients that are being verified or already in use in modern medicine and veterinary are also examined.