[The antibiotic sensitivity of Corynebacterium diphtheriaе gravis tox+ in composition of mixed biofilms.]

The purpose of study is to determine sensibility of strains of C. diphtheriae gravis tox+ in the composition of mixed biofilms to antibacterial medications. The object of study was strains of Corynebacterium diphtheriae gravis tox+ № 665, Corynebacterium pseudodiphtheriticum (poly-resistant), Corynebacterium pseudodiphtheriticum (antibiotic-resistant). Two mixed biofilms (120- and 720-hours) of strain of Corynebacterium diphtheriae gravis tox+ № 665 with strains of Corynebacterium pseudodiphtheriticum (poly-resistant), Corynebacterium pseudodiphtheriticum (antibiotic-resistant) were formed. The sensibility of typical and biofilm cultures of Corynebacteria to antibiotic medications was established using technique of serial dilution in fluid growth medium on the basis of MPC (mg/l). The typical culture of museum strain C. diphtheriae gravis tox+ № 665, modeling mixed biofilms had a sensitivity to all used antibacterial medications. The analysis of antibiotic resistance of strain of C. diphtheriae gravis tox+ № 665 in content of biofilm with strain С. pseudodiphtheriticum, poly-resistant to four antibacterial medications (cefazolin, kanamycin, azithromycin, ciprofloxacin, IPC from 1.25±0,00 to >5±0,00 mkg/ml) set reduction (p≤0,05) sensitivity 120-hour biofilm cultures of this strain to cefazolin, azithromycin, ciprofloxacin (MIC of 0, 234 and up to ±0,11>5±0,00 ug / ml) established decreasing (p≤0,05) of sensitivity of 120-hours biofilm culture of this strain to cefazolin, azithromycin, ciprofloxacin (MIC from 0, 234±0,11 to >5±0,00 mkg/ml) and 720-hours culture - to all mentioned medications (MIC from 0,312±0,00 to >5±0,00 mkg/ml). The comparison of antibiotics resistance of biofilm cultures of strain C. diphtheriae gravis tox+ № 665 established that indices of MIC of most of antibacterial medications and especially to those that poly-resistant and forming biofilm strain С. рseudodiphtheriticum was resistant, were higher as related to biofilm cultures of diphtheritic bacteria separated from biofilm with antibiotic resistant strain С. рseudodiphtheriticum. The trend was established related to decreasing of antibiotic sensitivity of Corynebacteria at cultivation in content of mixed biofilms with representatives of opportunistic Corynebacteria having polyresistance to antibacterial medications.

[THE IMMUNOLOGIC INDICATORS IN PATIENTS WITH CARIES OF CONTACT SURFACES OF LATERAL TEETH].

The samplings of patients aged 18-45 years with caries of contact surfaces of lateral teeth (n=18) and healthy adults aged 18-20 years with intact teeth (n=18) were examined The saliva taken in rotary vial on empty stomach in amount of 3-4 mi served as assay for analysis. To identify secretory immunoglobulin A, interleukin lß, interleukin-4 and interferon y saliva was centrifuged during 15 min under 1500 rpm. The supernatant fluid was analyzed using enzymoimmunoassay (test-systems "Vector-Best", Novosibirsk). The registration of reaction was implemented using multiscan Labsystem under wavelength 450 nm. The content of secretory immunoglobulin A was expressed in mg/l, cytokines - in pg/ml. It is demonstrated that in patients with caries average level of interleukin lß was almost two times higher (p<0.05) than analogous indicator in healthy examined patients. In healthy patients average level of interferon γ significantly (more than in 10 times) exceeded upper limit of allowable standard and was higher (p<0.05) in comparison with such in patients with caries of contact surfaces. The analysis of content of secretory immunoglobulin A in saliva established that in healthy patients average values of the given indicator were higher (p<0.05) than in patients with caries of contact surfaces of lateral teeth. The lower content of secretory immunoglobulin A and interferon y against the background of increased level of interleukin lß was detected in saliva of patients with caries of contact surfaces of lateral teeth. This occurrence can be considered as factor predisposing to development of caries process.

[THE DIAGNOSTIC APPROACHES TO VERIFICATION OF STREPTOCOCCUS INFECTION IN PATIENTS WITH INFECTIOUS MONONUCLEOSIS].

The Rostovskii state medical university of Minzdrav of Russia, 344022 Rostov-on-Don, Russia The analysis is applied concerning significance of laboratory techniques of verification of streptococcus infection (bacteriological analysis, detection of anti-streptolysin O in pair serums) in 148 patients with infectious mononucleosis aged from 3 to 15 years. The content of anti-streptolysin O exceeded standard in 41 ± 4.8% of patients with concomitant in acute period and in 49.5 ± 4.9% during period of re-convalescence. This data differed from analogous indicator in patients with negative result of examination on streptococcus infection independently of period of disease (9.3 ± 2.8%). The exceeding of standard of anti-streptolysin O was detected more frequently (t ≥ 2, P ≥ 95%) in patients with isolation of Streptococcus pyogenes (56.9 ± 5.8%) than in patients with Streptococcus viridans (31.2 ± 6.5%). The concentration of anti-streptolysin 0 in patients with concomitant streptococcus infection varied within limits 200-1800 IE/ml. The minimal level of anti-streptolysin O (C = 200 IE/mI) was detected independently of type of isolated Streptococcus and period of disease. The high levels of anti-streptolysin O were observed exclusively in patients with isolation of Streptococcus pyogenes. In blood serum ofpatient with concomitant streptococcus infection (Streptococcus pyogenes + Streptococcus viridans) increasing of level of anti-streptolysin O was detected in dynamics of diseases from minimal (C = 200 IE/ ml) to moderately high (200 < C < 400 IE/mI). It is demonstrated that to identify streptococcus infection in patients with infectious mononucleosis the anamnesis data is to be considered. The complex bacteriological and serological examination ofpatients is to be implemented This is necessary for early detection ofpatients with streptococcus infection and decreasing risk of formation of streptococcus carrier state.

[Mechanisms of formation of post-vaccinal immune response in children immunized with APDT and ADT-M preparations].

AIM: Study the mechanisms of formation of cell and humoral immunity against pertussis, diphtheria and tetanus in children immunized with immunobiological preparations (APDT vaccine and ADT anatoxin). MATERIALS AND METHODS: 30 practically healthy children (6 - 9 years of age) immunized with APDT and ADT-M preparations had TLR2, TLR4 expression determined in mononuclear cells (MNC). Vaccine preparations (APDT, ADT-M, AD-M, AT) and Corynebacterium diphtheriae gravis tox+, C. diphtheriae mitis tox- and Bordetella pertussis 345 were used as ligands. Cytokine production was determined in EIA. Content of anti-diphtheria, anti-tetanus and anti-pertussis antibodies--by PHA reaction and EIA. RESULTS: During stimulation with vaccines and B. pertussis 345 strain MNC were characterized by an increase (p < 0.05) of expression level of TLR2 and TLR4 and did not respond to stimulation with C. diphtheriae gravis tox+ and C. diphtheriae mitis tox- strains. Similar results were obtained during study of cytokine production (TNFalpha, IL-1, IL-6). A direct correlation between levels of antitoxic antibodies against diphtheria and tetanus (R = 0.486), antibacterial antibodies against pertussis and diphtheria was detected (R = 0.529). CONCLUSION: Analysis of cytokine production profile and determination of surface TLR expression can be used during evaluation of functional status of innate immunity cells and intensity of post-vaccinal immunity.

[The ability of diphtheria causative agent to form biofilm].

The article deals with results of studying diphtheria causative agent capacities to form biofilm as one of mechanisms of persistence in human organism. The study object was strain of C.diphtheriae gravis tox+ obtained from nasopharynx of patient aged 19 in municipal hospital No 1 of town of Gukovo of Rostov oblast in 2011. The patient had diagnosis of "diphtheria of nasopharynx, typical filmy, localized, mild severity, even course". The control was implemented using the museum strain C.diphtheriae gravis tox+ No 665 from the L.A. Tarasevitch state research institute of standardization and biologic preparations control. It is established that diphtheria causative agent as an ability to form biofilm. The intensity of process of formation of exopolysaccharide is higher on glass that on plastic surfaces. The differences in degree of intensity of formation of biofilm are revealed between the strain circulation in population and museum strains C.diphtheriae gravis tox+. The vital capacity of biofilm forming microorganisms is related with adaptation possibilities of strains.

[State of immunity to diphtheria and tetanus in women in early postpartum period].

AIM: Study of anti-diphtheria and anti-tetanus immunity in women in early postpartum period depending on age. MATERIALS AND METHODS: Women in early postpartum period (n =139) with unknown vaccine anamnesis aged 17 to 44 years and under the supervision of Rostov-on-Don maternity hospital No. 2 were examined for the evaluation of the anti-diphtheria and anti-tetanus immunity state. RESULTS: All the women had high level of protection form these infections. The level of anti-tetanus immunity intensity in the examined was higher than anti-diphtheria. CONCLUSION: Monitoring of anti-diphtheria and anti-tetanus immunity in women of childbearing age is necessary to resolve the issue of vaccine administration in this group. High level of maternal immunity intensity will allow to form a sufficient protection from infectious agents in neonates.

[Immunoenzyme technique for detection of salivary anti-diphtheria antibacterial antibodies].

Enzyme immunoassay for determining salivary anti-diphtheria antibacterial IgA was developed to evaluate local anti-diphtheria immunity. One hundred and thirty-one apparently healthy preschool and school children aged 2 to 16 years, inoculated with diphtheria and tetanus toxoids and pertussis and attenuated diphtheria and tetanus toxoid vaccines by the immunization schedules. The apparently healthy children's saliva were found to have anti-diphtheria antibacterial sIgA, the mean content of which in the examinees aged 2-16 years was 49.4 +/- 3.0 units/ml and depended on age. The level of antibacterial anti-diphtheria sIgA significantly increased in children by age 7-9 years and unchanged by 16 years of age. Comparison of sIgA levels in children with diphtheria morbidity could identify 2-6-year-old children as a diphtheria infection risk group.