A nutrient medium for the isolation and cultivation of Campylobacter

Campylobacter agar, nutrient medium intended for the isolation of bacteria of the genus Campylobacter from clinical material, has been developed. The composition of the medium includes sprat hydrolysate, aerotolerant additive (ferric sulfate--oxide, sodium pyruvate, sodium pyrosulfite), sodium glutaminate, agar. The selective properties of the medium are ensured by introducing the mixture of antibiotics consisting of polymyxin B, rifampicin, amphotericin B, ristomycin. The balanced composition of Campylobacter agar ensures the aerotolerance of Campylobacter organisms and gives the optimal conditions for their growth when the inoculated material is cultivated in the atmosphere made up of the mixture of three gases (5% of oxygen, 10% of carbon dioxide, 85% of nitrogen), as well as under the conditions of a "candle vessel". The medium suppresses the development of the associative microflora diluted 10(-1). As shown in the trial of the quality of Campylobacter agar by the inoculation of material taken from patients with acute enteric infections, agricultural animals and monkeys, the medium has pronounced selective, properties with regard to extraneous microflora, while ensuring the isolation of Campylobacter on the level of the control medium.     

Development and diagnostic value evaluation of a nutrient medium for the yield and isolation of hemoculture

The effectiveness of the bacteriological study of the blood depends in many aspects on the quality of nutrient media. The currently approved nutrient media for the isolation and yield of hemocultures have low isolation rates and require prolonged time for the isolation of causative agents. A two-phase nutrient medium for the yield and isolation of hemoculture has been developed. This medium has been found to have advantages over the control "double medium" in sensitivity, yield effect, growth rate and isolation rate with respect to the causative agents of bacteriemia and sepsis. The medium is standard and ready for use. The above-mentioned advantages of the medium increase the quality of the bacteriological diagnosis of bacteriemia and sepsis.     

Nutrient medium for the isolation and cultivation of pneumococcus

A culture medium for the isolation and cultivation of pneumococci, produced in a solid or liquid form and based on raw material unsuitable for use as foodstuff (human placenta), has been developed. The amino acid composition of the medium has been studied. The medium has been found to contain 19 amino acids, to be free from ballast serum proteins and blood, and to ensure the good growth of pneumococci isolated from pathological material, the formation of the normal capsule, as well as active biological properties. The medium has proved to create elective and selective conditions enhancing the effectiveness of investigations and simplifying the isolation of pneumococci in the microbiological examination of patients.     

Selective nutrient medium for the isolation of Bacillus cereus

A selective dried medium for the isolation of B. cereus from clinical material and foodstuffs has been developed. The medium has high selective properties which ensure the isolation of B. cereus from microbial association in pure culture, thus making it possible to accelerate further identification of the microorganisms.     

The intensity of cell receptor catabolism in intestinal yersiniosis and pseudotuberculosis

The intensity of the catabolism of cell receptors in yersiniosis was evaluated by the level of R-proteins in the blood sera of 95 patients. The study revealed that the degree of an increase in the level of R-protein depended on the severity and the course of the pathological process was the same in both nosological forms of the disease. The high and stable level of R-proteins was indicative of the possibility of the prolonged and relapsing course of yersiniosis.     

A dried nutrient medium for the isolation of Vibrio cholerae

A dried differential nutrient medium for the isolation of V. cholerae has been developed. The medium is sufficiently sensitive, has pronounced differentiating properties and greatly inhibits the appearance of microbial associations. During the cultivation of V. cholerae with the use of this medium the cultural, morphological and agglutination properties of the initial strains are retained.     

Inclusion of geothermal water in composition of nutrient medium for mycobacteria cultivation

New effective nutrient medium for isolation and cultivation of mycobacteria was developed. Time of their growth on the developed medium decreased by 3 to 11 days compared with control media (Finn II, Levenstein-Jensen). Rate of mycobacteria isolation increased by 20 to 40% compared with control media.     

A novel selective medium for isolation of Streptococcus mutans

The aim of this study was to improve the selective medium of Streptococcus mutans. A new selective medium, designated MS-MUTV, was prepared by adding 10 mg/l valinomycin to the MS-MUT medium previously described. The average recovery of S. mutans was 72.1%, and the growth of S. sobrinus and S. anginousus group was inhibited on MS-MUTV, but allowed on MS-MUT. One hundred and thirty-nine human saliva samples were examined and counted for S. mutans and non-S. mutans colonies. The recovery of S. mutans on MS-MUTV was similar to that on MS-MUT. Eighty-two and 7.9 percent of the saliva samples obtained S. mutans pure cultures, with no bacterial growth on MS-MUTV, respectively. The remaining 10.1% were contaminated with non-S. mutans, with low-level CFU. MS-MUTV is useful for the isolation of S. mutans alone from clinical samples in routine examinations.     

Viability of escherichia coli cells under long-term cultivation in a rich nutrient medium

We investigated the viability of Escherichia coli cells during long-term cultivation in Brain Heart Infusion (BHI) medium and observed that the number of viable cells increased, then decreased, and increased again, in this medium, and finally the cells died out within about 10 days. This cell death may result from an increase in the pH of the medium. After repeated cultivation in BHI, bacterial cells that did not die out even under conditions of further cultivation were obtainable from cultures showing a stabilized viable count. We propose that long-term cultivation in BHI medium is a good system for studying growth phase-specific events in E. coli cells, because the total life-cycle of a population of E. coli, including exponential growth, stationary phase, and extinction, can be seen during a period of only about 10 days. Also, this system clearly allows detection of a phenotype that may not be detectable in other commonly used media. Moreover, in this report, we show that mutants displaying the GASP (growth advantage in stationary phase) phenotype appear at high frequency under long-term cultivation conditions.     

Prions are novel infectious pathogens causing scrapie and Creutzfeldt-Jakob disease

Scrapie and Creutzfeldt–Jakob disease (CJD) are caused by prions, which appear to be different from both viruses and viroids. Prions contain protein which is required for infectivity, but no nucleic acid has been found within them. Prion proteins are encoded by a cellular gene and not by a nucleic acid within the infectious prion particle. A cellular homologue of the prion protein has been IDentified. The role of this homologue in metabolism is unknown. Prion proteins, but not the cellular homologue, aggregate into rod-shaped particles that are histo-chemically and ultrastructurally IDentical to amyloid. Extracellular collections of prion proteins form amyloid plaques in scrapie- and CJD-infected rodent brains as well as CJD-infected human brains. Within the plaques, prion proteins assemble to form amyloid filaments. Elucidating the molecular differences between the prion protein and its cellular homologue may be important in understanding the chemical structure and replication of prions.     

Antibiotic sensitivity of beta-hemolytic Streptococcus group A on a new nutrient medium for the cultivation of streptococci

Sensitivity of 167 strains of beta-hemolytic streptococci of group A was studied with the method of serial dilutions on a solid agar medium for cultivation of streptococci. The medium was developed at the I. I. Mechnikov Research Institute of Vaccines and Sera. It does not require addition of blood or serum. The strains were found to be highly sensitive to penicillin, cephalothin and erythromycin. The number of the strains resistant to tetracycline, streptomycin, gentamycin, levomycetin (chloramphenicol) and ristomycin amounted to 51, 36, 23, 1.8 and 1.8 per cent, respectively. One of the strains (0.6 per cent) was resistant to lincomycin. Strains with multiple resistance were isolated. The necessity of regular control of distribution of antibiotic resistance among staphylococci is indicated.     

Analysis of the plasmid composition of Yersinia pseudotuberculosis strains and its use for typing pseudotuberculosis pathogens

Electrophoresis in agarose gel has been used to study the plasmid spectra of 854 Yersinia pseudotuberculosis strains isolated from different sources. The plasmids found in the microbial strains are represented by the elements with molecular masses 82; 57; 45; 5.5; 4.4; 3.5; 2.7; 2.4; 2.3 Md. The variable spectra of plasmids is peculiar only for serovar I of Yersinia pseudotuberculosis. Plasmids p45 and p82 are classified as the main, while other plasmids as auxiliary ones. In accord with the classification all plasmid containing strains are divided into 8 plasmid strains. Using the proposed method for intraspecific typing of Yersinia pseudotuberculosis permits one to perfect the epidemiological analysis of pseudotuberculosis infection and make concrete the direction of prophylactic and antiepidemic measures.     

Direct isolation of specific RNA-interacting proteins using a novel affinity medium

Isolation of proteins that specifically interact with a given RNA or RNA regulation element is essential for studies on the molecular mechanisms of gene expression. Here, a novel method for direct isolation of such interacting proteins is described. It uses an affinity medium that consists of an interacting RNA with an artificially added ‘tail’, which is annealed to one end of a DNA ‘arm’, the other end of which is fixed covalently on the surface of aminosilanized glass powder. Thus the RNA itself is fully suspending, facilitating its interactions with proteins in its natural conformation. The proteins bound on the interacting RNA are eluted and subjected to SDS–PAGE, and the Coomassie-stained protein bands are cut and subjected to mass spectrometry (MS) analysis. Using this method, three proteins specifically interacting with the C/EBPβ 3′-untranslated region (3′-UTR) RNA were isolated and identified. This method is simple and convenient, and the DNA-glass powder medium can be used repeatedly.     

Development of novel Alicyclobacillus spp. isolation medium

AIM: To develop a new isolation medium with higher recovery rates of Alicyclobacillus spp. METHODS AND RESULTS: SK agar was developed with optimized incubation temperature, pH, acidulant, Tween 80 concentration and divalent cation addition. Results indicate that detection of Alicyclobacillus spp. by SK agar was significantly higher (P > 0.05) than those obtained by K agar, orange serum agar, and potato dextrose agar. CONCLUSIONS: Current media used for Alicyclobacillus spp. isolation still resulted in high numbers of false negative products. The sensitivity of SK agar to Alicyclobacillus spp. allows detection of low numbers of Alicyclobacillus spp. and also provides a more higher isolation results compared with currently used media. SIGNIFICANCE AND IMPACT OF THE STUDY: SK agar will be useful to the fruit juice industry to obtain more accurate numbers of contaminant Alicyclobacillus spp. With this media, false negative samples can be reduced, and the likelihood of exported products being rejected can be greatly reduced.     

A novel serum-free medium for the cultivation of Vero cells on microcarriers

Summary A novel serum-free medium for the cultivation of Vero cells on microcarriers was developed,which composed of the 1:1 mixture of Dubecco's Modified Eagle Medium: Nutrient Mixture F12, bovine serum albumin(BSA) or human serum albumin(HSA), epidermal growth factor(EGF), gelatin and Dbiotin. Both BSA and EGF were effective on cell growth, adhesion and spreading. Further addition of gelatin and biotin led to the enhanced cell adhesion and spreading without growth promoting activity. The serum-free medium was suitable for the cultivation of vero cells on several different microcarriers with cell density reached over 3×l0⁶cells/ml.