[THE CHROMOGENIC SYNTHETIC MEDIUM "KLEBSIELLA 5-ASK CHROM-C" FOR ISOLATION AND IDENTIFICATION OF KLEBSIELLAE].

The chromogenic synthetic medium "Klebsiella 5-ASK CHROM-C was developedfor isolation and identification of klebsiellae of species of K. pneumoniae subsp. pneumoniae K. oxytoca, K. mobilis according chromogenic reaction to enzyme 5-aminosalycilate decarboxylase as a unique marker of genus Klebsiella. The L-proline and L-calcium glutamate are used as a source of nitrogen and carbon in medium. The consistency of composition of growth medium that ensure its regularity. The diagnostic sensitivity of chromogenic medium is 95.3 ± 1.7%; diagnostic specificity is 100%; analytical sensitivity is 1-2 colony-forming units per ml-1. The identification of Klebsiella is achieved simultaneously with their isolation during 24 = 48 hours. The test of 5-ASK decarboxylase using two chromogenic mediums "Klebsiella 5-ASK CHROM-C" permits identifying additionally K. pneumoniae subsp, ozaenae, K .pneumoniae subsp. Rhinoscleromatis.

[The identification of clinical strains Pseudomonas fulva using techniques of MALDI-TOF mass spectrometry and common analysis].

The technique MALDI-TOF mass spectrometry was applied using device Microflex with database MALDI Biotyper (Bruckeer Daltonics Inc.) to identify with high level of reliability 8 strains P. fulva from collection of pseudo monads isolated from clinical material in St. Petersburg. When analyzing the same strains applying technique MALDI TOF mass spectrometry using device Vitek MS (bioMerieux) these starins were wrongly identifies as P. putida. The complex of tests of common analysis was approved and proposed for control differentiation of P. fulva and P. putida. The medical significance of P. fulva was approved.

[The synthetic growth medium Kings BS for detection of synthesis of fluorescein by bacteria Pseudomonas].

The synthetic growth medium KINGS BS was developed on the basis of substitution of proteose-peptone by L-glutamine or L-glutamine acid. The medium is used to detect the synthesis of fluorescein by bacteria Pseudomonas. The actual medium has equal diagnostic sensitivity and specificity as compared with medium King B. At the same time, the medium excels King B by content regularity and has no need of sterilization in vapor sterilizer and detects fluorescein in strains producing other pigments. The medium is suitable for standardization of technique of detection of synthesis of fluorescein by pseudomonads.

[Isolation of Shewanella algae from pleural exudate of patient with pneumonia].

AIM: Clinical-microbiological description of the first case in Russia of isolation of S. algae bacteria from clinical material. MATERIALS AND METHODS: Patient P., 23 years of age, diagnosis: right-sided distal pneumonia, severe course; parapneumonic empyema of pleura. Bacteria isolation, cultural and biochemical tests differentiating S. algae and Shewanella putrefaciens were performed according to Holt H.M. et al., 2005. Identification of bacteria, tests of sensitivity to antibiotics were carried out by automatic system Vitek 2 (bioMerieux) and additionally by disc-diffusion method. RESULTS: S. algae in association with Serratia marcescens were isolated from pleural exudate of the patient with pneumonia. S. algae bacteria had typical taxonomical features and pathogenicity factors (lipase, gelatinase, beta-hemolysin); were resistant to benzylpenicillin, amoxicillin, cefazolin and sensitive to other beta-lactam antibiotics, aminoglycosides, fluoroquinolones, macrolides, tetracycline. CONCLUSION: S. algae bacteria isolated from pleural exudate of the patient with pneumonia are etiologically significant in parapneumonic empyema of pleura.

[Pyrazinamidase activity of bacteria from Enterobacteriaceae family as characteristic for taxonomy].

Pyrazinamidase activity in 330 strains of bacteria from Enterobacteriaceae family (14 genus, 27 species) has been assessed. Pyrazinamidase activity detected in species from following genuses: Citrobacter, Escherichia, Klebsiella, Kluyvera, Morganella, Providencia, Raourtella, Salmonella, Shigella, and also in Proteus mirabilis, and nonpathogenic serovars of Yersinia enterocolitica, Y. frederiksenii. Pirasinamidase was absent in Serratia (S. marcescens, S. liguefaciens), Hafnia alvei, P. vulgaris, P. penneri, Y. pseudotuberculosis and pathogenic serovars of Y. enterocolitica. Absence of pyrazinamidase activity in bacteria from Hafnia and Serratia genus is a key taxonomic characteristic for identification of enterobacteria with microvolume assay technology.

[Profiles of the utilization of 20 amino acids as the only source of nitrogen and carbon in bacteria of the genera Klebsiella, Enterobacter, Serratia, Escherichia].

The profiles of the utilization of 20 protein amino acids in 118 Klebsiella pneumoniae sub- sp. pneumoniae, K. oxytoca, K. planticola, K. mobilis, Enterobacter cloacae, Serratia marscescens, S. liquefaciens, Escherichia coli strains isolated from clinical material were studied. The utilization of amino acids was determined on minimal saline agar containing amino acid as the only source of nitrogen and carbon; the results were evaluated after 72-hour incubation at 37 degrees C. 17 profiles of amino-acid utilization were thus determined, most of them genus-specific in enterobacteria: Klebsiella (profiles No. 1--6, 9, 10), Enterobacter (No. 11--13), Serratia (No. 14--16), Escherichia (No. 17). The full coincidence of amino-acid utilization profiles in bacteria of K. mobilis (No. 1, 6) and K. pneumoniae subsp. pneumoniae with out of such profiles in bacteria of the genera Enterobacter, Serratia, Escherichia was established, which confirmed that K. mobilis (formerly Enterobacter aerogenes) belonged to the genus Klebsiella.

[Antibiotics sensitivity and characteristics of the esculin-positive Pseudomonas aeruginosa biovar]. / Chuvstvitel'nost' k antibiotikam i svoistva éskulinopozitivnogo biovara Pseudomonas aeruginosa.

Strains of Pseudomonas aeruginosa hydrolyzing esculin were isolated for the first time. They amount to 17.1 +/- 2.0% (60 from 325) of the investigated P. aeruginosa strains isolated from the clinical material in St. Petersburg. Esculin hydrolysis was measured by micromethod in plates, results were analysed after 3-hours incubation at 37 degrees C. Esculin-positive strains possesed biovar properties: they are widely spread, demonstrated other characteristic features (absence of triethylamine odour, specific colonies lysis), are stable on ability to hydrolyse esculin while culture storage and after repeated culturing. Typical strain of esculinolytica biovar was deposited into the culture collection of the National Research Institute of Agricultural Microbiology as P. aeruginosa ARRIAM 64-A. Susceptibility testing of the esculin-positive strains by disk-diffusion method revealed that most strains were inhibited by imipenem (86.6%), amikacin (75.0%), ceftazidime (65.0%), meropenem (60.0%), aztreonam (51.6%). The percent of strains susceptible to other antibiotics was lower: azlocillin--33.3%, netilmycin--33.3%, piperacillin--26.6%, ceftriaxon--18.3%. Only small number of strains were inhibited by ciprofloxacin (8.3%), gentamycin (3.4%), cefoperazone (1.7%) and carbenicillin (1.7%). The results may be used for empiric therapy before the isolated strain susceptibility is tested but only according to positive esculin-hydrolysis express-test evaluated in 3-hours period.

[The description of an esculin-positive biovar of Pseudomonas aeruginosa]. / Opisanie éksulinpozitivnogo biovara Pseudomonas aeruginosa.

In the study of 280 P. aeruginosa strains isolated in different hospitals of St. Petersburg for the first time 48 strains capable of hydrolyzing esculin have been detected. The hydrolysis of esculin is determined in plates with the use of the microvolume techniques the results were evaluated after 3-hour incubation at 37 degrees C. The data confirming the existence of the exculin-positive biovar of P. aeruginosa have been obtained; these data show the wide spread of esculin-positive strains in hospitals of different specialization (17.1 +/- 5.1% of P. aeruginosa strains), the characteristic combination of the sign of esculin hydrolysis with such signs as the absence of the smell of trimethylamine and the phenomenon of "iridescent lysis" of the colonies, the stability of the sign of esculin hydrolysis in strains, repeatedly isolated from patients, after the storage of the cultures and their treatment with plasmid-eliminating preparation. The name "esculinolytica" has been proposed for this biovar. The typing strain of biovar esculinolytica has been deposited in the culture collection of the Russian Research Institute of Agricultural Microbiology as P. aeruginosa ARRIAM 64-A. This biovar been found to be most widely spread in urological hospitals, where esculin-positive strains are isolated 3 times more frequently (32.2 +/- 5.1% of P. aeruginosa strains) than in surgical hospitals (10.7 +/- 2.2%).

[Mechanism of specific chromogenic reaction by Klebsiella spp. on nutrient medium with 5-aminosalicylic acid]. / Mekhanizm spetsificheskoi khromogennoi reaktsii Klebsiella spp. na pitatel'noi srede s 5-aminosalitsilovoi kislotoi.

The nature of the factor and the mechanism of color reaction with 5-aminosalicylic acid (5-ASA) inherent in bacteria of the genus Klebsiella were investigated. The color reaction was shown to proceed in two stages. During the first stage, occurring in aerobic and anaerobic conditions, the tested strains of Klebsiella decarboxylize 5-ASA yielding CO2 and p-aminophenol, which is a colorless product. During the next strictly aerobic stage which proceeds without participation of bacteria or their exoenzymes, p-aminophenol is oxidized by the air oxygen forming the dark-brown polymers. The color reaction shows high substrate specificity. It is suggested that the color reaction is realized by a previously unknown lyase-type enzyme 5-aminosalicylate decarboxylase (5-aminosalicylate-carboxy-lyase) localized inside the Klebsiella cells.

[A unique marker of Pseudomonas from the first group of rRNA homology--selective sensitivity to the bacteriostatic action of barium ions]. / Unikal'nyi priznak psevdomonad pervoi gruppy rRNK-gomologii--izbiratel'naia chuvstvitel'nost' k bakteriostaticheskomu deistviiu ionov bariia.

On the basis of the study of the bacteriostatic action of chlorides and nitrates of barium and 24 other metals on 18 Pseudomonas species of all groups of rRNA homology and on 49 bacterial species belonging to 25 other genera, unique selective sensitivity to the bacteriostatic action of Ba2+ ions has been established in Pseudomonas of the first group of rRNA homology. The marker of barium sensitivity is in line with changes in the classification of Pseudomonas and is of interest for their further more precise systematization. The test for barium sensitivity of bacteria helps simplify the identification of Pseudomonas of the first group of rRNA homology and makes the identification more accurate.

[The hydrogen peroxide microvolumetric method of the OF test: the rapid determination of glucose oxidation and fermentation by gram-negative bacteria]. / Peroksidvodorodnyi mikroob''emnyi metod OF-testa: ékspressnoe opredelenie okisleniia i fermentatsii gliukozy gramotritsatel'nymi bakteriiami.

For the first time the method of the rapid (1 hour) screening of groups of gram-negative bacteria by the OF test with glucose, carried out with the use of microvolumetric techniques, has been developed. The method is based on the use of hydrogen peroxide at non-bactericidal concentration as a component of a liquid buffer medium containing indicator and glucose and intended for the oxidation of glucose. Catalase of bacteria introduced into the medium for study ensures the rapid saturation of the medium with oxygen and the completion of the oxidation of glucose in 10-60 minutes. An equal period is necessary to achieve the complete fermentation of glucose in the same medium without hydrogen peroxide. The method has been proved to yield significant results in the joint study of the OF test on 502 strains, belonging to 21 genera of fermenting and nonfermenting gram-negative bacteria, by the hydrogen peroxide method and in Hugh-Leifson medium.

[Rapid determination of nitrate reductase in non-fermenting gram-negative bacteria]. / Uskorennoe opredelenie nitratreduktazy nefermentiruiushchikh gramotritsatel'nykh bakterii.

Experimental study of 22 types of nonfermenting gram-negative bacteria, carried out in 317 strains, has demonstrated a high accuracy and reproducibility of the rapid (taking but 3 hrs) measurement of nitrate reductase in these bacteria by a microtest in culture medium with 0.1 percent of NaNO3 in plates; 0.2 percent solution of rivanol and 12 percent hydrochloric solution are employed as the reagents for nitrates.

[Pseudomonas APS nutrient medium for the isolation and identification of Pseudomonas aeruginosa and Pseudomonas putida]. / Pitatel'naia sreda "Pseudomonas APS" dlia vydeleniia i identifikatsii bakterii Pseudomonas aeruginosa i Pseudomonas putida.

Pseudomonas APS selective medium has been developed on the basis of a newly detected selective antibacterial action of oxaphenamide (p-oxyphenylsalicylamide), a cholagogue. This medium permits a single-stage combined isolation and identification of P. aeruginosa after 16-24 hrs incubation of inoculated material at 42 degrees C. If the material is incubated at 35-37 degrees C, isolation of P. putida and P. aeruginosa is possible, that are differentiated by a nitroreductase microtest within 3 hrs. The sensitivity and selective ability of the developed medium are superior to other media manufactured in this country and commercial media with cetrimide and irgasan, produced abroad.

[A specific property of bacteria in the genus Klebsiella--a color reaction with 5-aminosalicylic acid in the nutrient medium]. / Spetsificheskoe svoistvo bakterii roda Klebsiella--tsvetnaia reaktsiia s 5-aminosalitsilovoi kislotoi v pitatel'noi srede.

For the first time bacteria of the genus Klebsiella have been found to possess a specific property, characteristic of this genus only, i.e. the capacity of giving color reaction with 5-aminosalicylic acid. This reaction can be observed in all Klebsiella species under study: K. pneumoniae (94.4 +/- 2.3%), K. ozaenae (93.3 +/- 4.5%), K. rhinoscleromatis (100%), K. oxytoca (88.0 +/- 4.9%), K. mobilis (92-5 +/- 4.3% of the strains). In all other bacteria under study (40 species, 22 genera and 7 families) the reaction is negative. The test for the color reaction with 5-aminosalicylic acid confirms the belonging of K. mobilis (Enterobacter aerogenes) to the genus Klebsiella, thus making it possible to simplify and accelerate the identification of Klebsiella.