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restored, and in 4 patients infiltration preserved in 2 patients were followed epidural sequesters and 2 months 31 patients activated with hard corset.
Conclusion:
Complex treatment of patients with inflammatory diseases of the spine complicated with epidural abscess through the use mik-
roirrigation tube on the site of the lesion prevents possible inflammatory complications in the postoperative period, allow recovery and active rehabilitation of patients and provides favorable conditions for seam and autograft reconstruction without resorption in the early postoperative period and significantly increasing their adaptation opportunities and quality of life.
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11. Peretsmanas E. O. Errors and difficulties in the diagnosis of tuberculous spondylitis./Peretsmanas E.O, Lavrov V.N, M. E. Saltykov//Sci. tr. Proc. scientific. Pract. Conf. - SPb. - 2006; 151.
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DOI: http://dx.doi.org/10.20534/ESR-16-9.10-49-51
Abdukhalilova Gulnara, Research Institute of Epidemiology, Microbiology and Infectious Diseases, kandidat of medical sciences Tuhtamurodov Hojiakbar, Tashkent Medical Academy, magistr Ibragimov Adil,
Research Institute of Epidemiology, Microbiology and Infectious Diseases, kandidat of medical sciences
Ahmedov Ildar,
Research Institute of Epidemiology, Microbiology and Infectious Diseases, kandidat of medical sciences Gulyamov Nariman Gylamovich, Research Institute of Epidemiology, Microbiology and Infectious Diseases, doctor of medical sciences professor
E-mail: saodat.us@mail.ru
Phenotypic and genotypic resistance to antimicrobial performance in stains of Salmonella Typhimurium
Abstract: We studied 39 strains of Salmonella Typhimurium, which were allocated from patients in 2012-2013 by bacteriological and molecular genetic methods, strains sensitivity to antimicrobial agents. It revealed an extremely high proportion of strains resistant to the antibiotic medicines, including — ^-lactams (84.6%) and fluoroquinolones (92.3%). Strains of S. Typhimurium characterized by multiple resistances to different classes of the antibiotics. About half of the isolates (43.6%) have the same resistance phenotype A-AC-T-C-CO-CTX-CA-NA-CPM, which may indicate the clonal propagation of the pathogen. Resistant strains of S. Typhimurium to ^-lactams (including cephalosporins of 3-4 generations) is due to produce ^-lactamase
Sectiom 6. Medical science
genetic families TEM and CTX-M, with 66,7% of the strains are characterized by the same genotype and resistance genes have four ^-lactamases: TEM, CTX-M1, CTX-M2 and CTX-M25, it can also indicate the clonal propagation of the pathogen. Keywords: antibiotic, resistance, gen, phenotype, genotype, S. Typhimurium.Introduction.
Antimicrobial resistance (the term "antibiotic" and "antimicrobial agents" in this document interchangeably) — a resistance to the action of a microorganism of the antimicrobial substance to which microorganism was previously sensitive. Resistance is not a new phenomenon and has been studied for the past 50-60 years. World Health Assembly in 1998 adopted the first resolution, directed at rational and efficient use of antimicrobial agents. In 2001 was published by the WHO strategy to contain antimicrobial resistance [1]. In 2011, WHO dedicated World Health Day the problem of antimicrobial resistance.
Excessive use of antibiotics in farm animals has serious consequences for public health, as it promotes the appearance of antibiotic resistant bacteria and resistance genes that can be transmitted to humans [7]. Resistance food related infections of zoonotic bacterial types Salmonella certainly associated with the use of antibiotics in farm animals; foodborne infection caused by such resistant bacteria repeatedly documented in humans. Of particular concern is the stability of the so-called "critically important antibiotics" used in medicine. For example, the use of fluoroquinolones in farm animals led to the appropriate antibiotic resistance in Salmonella which cause infectious diseases in humans. In diseases caused by multire-sistant strains of Salmonella typhimurium specific phage type (DT) 104 with resistance to quinolones, we observed treatment failure, a higher frequency of hospitalization and greater risk of death [5; 8].
The aim of the research are study of the sensitivity of antibiotic strains of Salmonella Typhimurium isolated from patients with acute enteric infections in Tashkent in 2012-2013.
Materials and methods.
The study included 39 strains of Salmonella Typhimurium isolated in 2012-2013 from patients with acute intestinal infection in the bacteriological laboratory of Scientific Research Institute Epidemiology, Microbiology and Infectious Diseases Ministry of Health of Uzbekistan, Tashkent, and sent for further examination to the "national collection of microbial infections of human" of Scientific Research Institute Epidemiology, Microbiology and Infectious Diseases Ministry of Health of Uzbekistan.
The study of morphological, biochemical and tinctorial properties of the strains of Salmonella spp. was performed according to WHO protocols [9], the determination of sensitivity to antimicrobial agents perfomed by disc — diffusion method and serial dilutions as recommended by CLSI M02-A11 and M07-A10 [10].Полученные результаты интерпретировали согласно рекомендациям CLSI M100-S22 [11]. The results are interpreted according to the recommendations CLSI M100-S22 [11]. We used Mueller-Hinton agar and disks with antibiotics production of Hi-Media, India, as well as chemically pure substance of antimicrobials (HIMEDIA, India). We studied the sensitivity of Salmonella to the following AMD: p-lactams (ampicillin, amoxicillin/clavulanic acid, cefotaxime, cefotaxime/clavulanic acid, ceftazidime, ceftazi-dime/clavulanic acid, cefepime, imipenem, meropenem), quinolones (nalidixic acid, ciprofloxacin, oflaksatsin), aminoglycosides (gentamicin) and other groups of drugs (tetracycline, chloramphenicol, trimethoprim-sulfamethoxazole). For internal quality control of the studies we have used E.coli ATCC 25922 and control strains K.pneumoniae ATCC 700603.
Mechanisms of resistance strains ^-lactam ILA studied by molecular genetic methods: PCR, electrophoretic detec-
tion and PCR in real time. Performed the detection of genes encoding products of ^-lactamase wide and spread spectrum genetic families SHV, TEM, CTX-M (genetic groups CTX-M1, CTX-M9 CTX-M8/25 CTX-M2), and various carbapenemases — OXA -48, KPC and metallo p-lactamase NDM, VIM, IMP. Isolation ofDNAby the method of Shevchenko O. V. et al. (3). For PCR amplification was used sets of reagents of production «Evrogen» (Moscow, Russia). The final reaction volume was 25 ul with the addition of template DNA. Reaction components were added according to the manufacturer's instructions. Amplification was performed according to the protocols [12; 2; 6; 3; 4]. The PCR products were visualized by electrophoresis (in 0.5X TBE buffer) in a 2% agarose gel containing ethidium bromide (electric field intensity at 6 V/cm for 30 min), and then view a UV transilluminator.
Results and discussion
Included in the research Salmonella strains isolated from patients with acute intestinal infections in the city of Tashkent in 2012 -. 2013, according to the culture-morphological, biochemical and antigenic properties consistent with S. Typhimurium. All studied strains were resistant to one or more classes of the AMD. Moreover, the proportion of strains resistant to critically important for medicine classes AMD (beta-lactams and fluoroquinolones) is extremely high. Thus, resistant to various p-lactam antibiotics (both unprotected and protected by clavulanic acid) of 84.6% (the spread spectrum cephalosporins) to 89.7% (ampicillin), the strains were characterized by very high stability — and MIC50 ampicillin and cefotaxime MIC 90 amounted to 512 mcg/ml ceftazidime 64 mg/ml and 128 mg/ml, respectively. In the group of p-lactam antibiotics have retained their activity against S. Typhimurium only carbapenems (imipenem, meropenem). Currently, the use offluoroquinolones for the treatment ofSalmonella infection is limited to the prevalence of strains that are insensitive (resistant or moderately resistant) to this group of drugs. Thus, 92.3% of subjects S. Typhimurium strains were resistant to nalidixic acid, while MIC50 and MIC90 amounted to 512 mcg/mL. Insensitive to fluoroquinolones (ciprofloxacin) strains accounted for 74.3% (moderately resistant — 69.2%, stable — 5.1%), the sensitivity retained 25.7%. It is noteworthy that the disc-diffusion method can not significantly determine the sensitivity of Salmonella strains to ciprofloxacin. When determining the sensitivity to this drug is necessary to use methods for determining MIC. According to the current international guidelines treatment with high probability fluoroquinolones Salmonella infection is ineffective in case the MIC for ciprofloxacin-pathogen strain greater than 0.06 mg/l. Our research revealed that MIC50 and ciprofloxacin MIC90 was 1 mg/ml, indicating a high probability of clinical inefficiency use of fluoroquinolones for the treatment of Salmonella infections in patients Tashkent.
Our research showed that among the studied strains S. Ty-phimurium 84.6% resistant to chloramphenicol, and MIK50 and MIC90 amounted to 512 mkg/mL. Also with the high frequency observed resistance to antibiotics tetracycline (97,4% resistant, MIC50 and MIC90-128 mg/ml), gentamicin (76.9% MIC50 = 64 mi-crograms/ml and MIC90 = 128 mcg/ml) co -trimocsasol (64.1% of strains MIC90 = 512 mcg/ml).
Research has shown that most of the investigated strains were characterized by combined multiple resistance to AMP.
Among dominated Salmonella strains tested multiple pheno-types with AMP resistance: 79.5% of the strains were characterized
by resistance 8 and more AMP, and combine these strains resistant to ^-lactams, products ^-lactamase a spread spectrum (PBLSS), with resistance to drugs other groups. The most frequently met phe-notype resistance to ampicillin, amoxicillin/clavulanic acid, cefotaxime, ceftazidime, cefepime, tetracycline, chloramphenicol, co-tri-moxazole, nalidixic acid (A/AC/CTX/CA/CPM/T/C/CO/NA), such strains accounted for 43.6%.
In S. Typhimurium strains resistant to ^-lactams, detection genes was performed, responsible for the production of ^-lactamases and extended broad spectrum of different genetic families.
The research did not reveal the genes encoding ^-lactamase production of a wide range of genetic and extended family the SHV, and carbapenemases genetic families OXA-48, RNC, VIM, NDM, IMP. Almost all studied strains of S. Typhimurium identified genes ^-lactamase TEM of genetic family (97.4% of the strains). Moreover, more than 80.0% of the strains had the ^-lactamase gene of genetic family CTX-M (92.3%), related to the genetic groups CTX-M 1 (87.2%), CTX-M 25 (87.2%) and CTX-M 2 (76.9%). The studied strains were characterized by a combination of.beta.-lactamase genes TEM and CTX-M of different genetic groups.
In 36 of the 39 investigated strains (92.3%) at the same time identified several genes ^-lactamases. Thus, more than 60% of the strains were predominantly genotype resistance genes including four p-lactamases: TEM, CTX-M1, CTX-M2 and CTX-M25
(27 strains, 69.2%). Eight strains (20.5%) had three genes simultaneously p-lactamase in various combinations, one strain — genes of two p-lactamase (TEM and CTX-M1). Two strains identified a gene p-lactamase (TEM). In one of strain S. Typhimurium ^-lactamases are not identified from above mentioned genetic families.
Thus, must be noted that in determining the sensitivity to fto-hinolonam necessary to use methods for determining the MIC of ciprofloxacin as a disco-diffusion method is not reliable enough.
Conclusions:
1. The population S. Typhimurium of strains isolated from patients with acute intestinal infections in Tashkent in 2012-2013, characterized by an extremely high percentage of strains resistant to the AMD, including critical to medicine — p-lactams (84.6%) and fluoroquinolones (92.3%).
2. Strains S. Typhimurium characterized by multiple resistance to different classes of the AMD. About half of the isolates (43.6%) have the same resistance phenotype A-AC-T-C-CO-CTX-CA-NA-CPM, which may indicate the clonal propagation of the pathogen.
3. Resistance of strains S. Typhimurium to beta-lactams (including cephalosporins 3-4 generations) is due to produce beta-lactamase genetic families TEM and CTX-M, 66.7% of the strains are characterized by the same genotype resistance and have four beta-lactamases genes: TEM, CTX-M1, CTX-M2 and CTX-M25, it can also indicate the clonal propagation of the pathogen.
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