Etiologic role of Corynebacterium non diphtheriae in patients with different pathology

Bacteriologic examination of 1589 patients showed that, aside from C. diphtheriae, 11% of acute upper respiratory tract infections were caused by other Corynebacterium species. Such bacteria can cause infections of various localizations (bronchitis, pyelonephritis, urethritis, colpitis, dermatitis, arthritis, etc.). C. pseudodiphtheriticum and C. xerosis were isolated from clinical specimens most frequently. Corynebacterium spp. have adhesive, hemolytic, hemagglutinating, and neuraminidase activity; some of them are highly pathogenic. The most virulent, were following species: C. diphtheriae, C. pseudotuberculosis, C. urealyticum, and C. ulcerans. Corynebacterium non diphtheriae were frequently isolated from clinical specimens in association with staphylococci and streptococci. In such cases, factors of pathogenicity and resistance to antibiotics were more pronounced. Strains isolated with association with other bacteria have lost susceptibility to tetracycline, oleandomycin, penicillin, and erythromycin. It is important to be vigilant about bacteria from Corynebacterium genus in clinical settings, and thoroughly study their biologic characteristics, especially in immunocompromised patients.     

Classification of coryneform bacteria associated with human urinary tract infection (group D2) as Corynebacterium urealyticum sp. nov.

Urealytic strains of coryneform bacteria that are designated Corynebacterium group D2 and are isolated from human urine are a cause of urinary tract infections. Cell wall and lipid analyses confirmed that these organisms are members of the genus Corynebacterium but can be separated from other species in the genus on the basis of DNA base composition and DNA-DNA hybridization values. Biochemically, strains in this taxon can be distinguished from other Corynebacterium spp. by their failure to produce acid from carbohydrates, by their failure to reduce nitrates, and by their ability to hydrolyze urea. We regard these bacteria as a new species of the genus Corynebacterium and propose the name Corynebacterium urealyticum. The type strain is strain NCTC 12011 (= ATCC 43042).     

Antibiotic resistance of coryneform bacteria isolated from the reproductive tract of women

The species composition, antilysozyme activity and antibiotic resistance of coryneform bacteria, isolated from the reproductive tract of women with microecological disturbances, were studied. Sixty six women without microecological disturbances and 102 female patients with microecological disturbances in the reproductive tract were examined. The study showed that Corynebacterium minutissimum, C. amycolatum, C. group JK, C. bovis and C. pseudodiphtheriticum prevailed in the healthy women. In the patients with microecological disturbances in the reproductive tract C. vitarumen, C. matruchotii, C. striatum, C. renale and C. urealyticum were detected in addition to the above species. The average antilysozyme activity of the coryneform bacteria isolated from the healthy women was 1.32 +/- 0.47. In the patients with microecological disturbances in the reproductive tract it was 1.84 +/- 0.38. The in vitro susceptibility of the coryneform bacteria to antimicrobial agents was determined. High prevalence of resistance to beta-lactams (oxacillin and penicillin), erythromycin and co-trimoxazole was detected. Thus, the species variety and the antilysozyme activity of the coryneform bacteria in the reproductive tract of the women with microecological disturbances were found to be higher. The high prevalence of resistance to oxacillin, penicillin, erythromycin and co-trimoxazole in the coryneform bacteria isolated from the patients with the microecological disturbances did not differ from that in the healthy women.     

Isoprenoid quinones in the classification of coryneform and related bacteria.

Menaquinones were the only isoprenoid quinones found in 85 of the 95 coryneform bacteria examined. Dihydromenaquinones having nine isoprene units were the main components isolated from Corynebacterium bovis, from other glutamic acid-producing strains, and from Arthrobacter globiformis and related species. Dihydromenaquinones with eight isoprene units were found in Brevibacterium linens, the remaining Corynebacterium species and strains probably belonging to the genus Rhodococcus. Tetrahydromenaquinones with eight isoprene units were found in Arthrobacter simplex and Arthrobacter tumescens, and with nine isoprene units in Cellulomonas and Oerskovia. Kurthia and Curtobacterium were characterized by menaquinones with seven and nine isoprene units, respectively, and Microbacterium lacticum and Corynebacterium aquaticum had comparable amounts of menaquinones with 10 and 11 isoprene units. Strains received as Brevibacterium leucinophagum, Corynebacterium autotrophicum, Corynebacterium nephridii, Mycobacterium flavum, Mycoplana rubra and Protaminobacter ruber contained uniquinones as their sole isoprenoid quinones. The isoprenoid quinone data correlate well with major trends in coryneform taxonomy and are of value in the classification of coryneform and related bacteria.     

A specific property of bacteria in the genus Klebsiella--a color reaction with 5-aminosalicylic acid in the nutrient medium

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.     

Reactivity to bacterial peptidoglycans in a phagocytosis system. 1. The range of immunological specificity of Staphylococcus aureus peptidoglycan

The antigenic features of S. aureus peptidoglycan (PG) were studied in the reaction of stimulation of oxygen-dependent neutrophil metabolism, mediated by the IgG opsonins of normal human serum. The study was carried out at different taxonomic levels: the species (S. aureus), the genus (Staphylococcus), the family (Micrococcaceae), as well as in relation to remote taxons (organisms belonging to the families Streptococcaceae, Enterobacteriaceae, Neisseriaceae, to the genus Corynebacterium). All S. aureus strains were identical with respect to the specificity of their PG, essentially differing from other bacteria in this regard. After the removal of antibodies to different PG the effectiveness of PG opsonization decreased by 10.4-44.7%. Such decrease was most pronounced in experiments with the PG of streptococci (S. pyogenes, S. faecalis, S. salivarius) and Micrococcus luteus.     

The microflora of ulzerous zone mucosa in patients with duodenal ulcer

Bacteriological study of the biopsies taken from gastric and duodenal mucosa of 10 healthy volunteers and 74 patients with duodenal ulcer, was carried out. In the gastroduodenal zone of healthy subjects microorganisms of 6 genera (Streptococcus, Candida, Staphylococcus, Bacillus, Helicobacter and Lactobacillus) were detected. H. pylori was isolated in 20% of cases only in biopsy specimens taken from the antral section of the stomach of healthy as monoculture or in combination with C. albicans. In patients with duodenal ulcer activation of opportunistic microflora was observed in the periulcerous zone. More often H. pylori occurred in associations with fungi of the genus Candida, streptococci, staphylococci, enterobacteria, Pseudomonas and other microorganisms (of more than 30 genera). Quantitatively the dominating microorganisms (3.8-5.7 lg CFU/g) were H. pylori, fungi of the genus Candida, bacteria of the genera Streptococcus, Peptostreptococcus, Bacteroides, Gemella, Prevotella, Veillonella, Peptococcus, Bacillus, different species of opportunistic enterobacteria, as well as bacteria of the genera Staphylococcus, Micrococcus, Corynebacterium, Neisseria, Pseudomonas, etc. Opportunistic bacteria detected in the ulcerous zone, as a rule, expressed hemolytic, lecithinase, RNAase, caseinolytic, catalase and urease activity. Sonicated filtrates of such cultures produced a cytotoxic effect on cells HEp-2. Ulcer is an infected wound that needs sanitation.     

Heavy metal effects on Proteus mirabilis Superoxide dismutase production

Abstract Levels of genomic DNA relatedness were determined using a SI nuclease procedure for reference bacteria identified as biotypes of Corynebacterium diphtheriae , biovars of Corynebacterium pseudotuberculosis , and ' Corynebacterium ulcerans '. These results showed that the three species are separate taxa at the genomospecies level whereas biotypes and biovars are closely related genomically within each species. Phylogenetic analyses of small-subunit rDNA sequences revealed that ' Corynebacterium ulcerans ' forms a tight cluster with Corynebacterium pseudotuberculosis within the robust branch that groups all Corynebacterium sequenced to date. Therefore, we propose that the species incertae sedis ' C. ulcerans ' should be conclusively recognized as a distinct species within the genus Corynebacterium with strain CCUG 2708 = NCTC 7910 as type strain. This species is characterized by urease production and fermentation of glycogen.     

Compatible solutes in representatives of the genera Brevibacterium and Corynebacterium: Occurrence of tetrahydropyrimidines and glutamine

Abstract The aim of our investigation was to study the haloadaptation of a number of species of Gram-positive bacteria belonging to the genera Brevibacterium and Corynebacterium . We used two different HPLC-techniques and ¹³C-NMR spectroscopy for the identification of osmolytes (compatible solutes). The tetrahydropyrimidines ('ectoines') are the main compatible solutes in the genus Brevibacterium , whereas accumulation of glycine-betaine and accumulation of synthesis of glutamine is mainly responsible for osmoadaptation in the genus Corynebacterium . Pipecolic acid, formerly described as a potential osmolyte synthesized de novo in C. ammoniagenes , does not contribute markedly to the solute pool, unless supplemented to the medium.     

Genome sequence of Corynebacterium nuruki S6-4 T, isolated from alcohol fermentation starter

Corynebacterium nuruki S6-4(T), isolated from Korean alcohol fermentation starter, is a strictly aerobic, nonmotile, Gram-positive, and rod-shaped bacterium belonging to the genus Corynebacterium and the actinomycete group. We report here the draft genome sequence of C. nuruki strain S6-4(T) (3,106,595 bp, with a G+C content of 69.5%).     

The use of batumin-containing disks for the rapid identification of staphylococci

The study of 467 microbial strains obtained from collections and from clinical sources revealed that microorganisms of the genus Staphylococcus were highly sensitive to batumin, a new antibiotic obtained from bacteria of the genus Pseudomonas. 378 strains of 15 Staphylococcus species proved to be highly sensitive to the diagnostic preparation "Diastaph", developed on the basis of batumin (antibiotic-impregnated discs); After 18-hour incubation the diameter of the growth inhibition zones on agar-containing culture media was 18-38 mm. Strains belonging to the genera Micrococcus, Dermacoccus, Kocuria and Kytococcus, as well as the tested representatives of other taxa (Planococcus, Streptococcus, Corynebacterium, Acinetobacter, Pseudomonas, Neisseria, the representatives of all tested genera of the family Enterobacteriaceae, fungi of the genus Candida) were insensitive to the diagnosticum. "Diastaph" permits not only the rapid identification of staphylococci pure cultures, but also the determination of their presence in association with other microbial species directly in pathological material, which makes it possible to recommend this diagnostic preparation for use in medical, veterinary and sanitary microbiology.     

Role of persisting opportunistic bacteria in the pathogenesis of the gastric and duodenum ulcer

Dynamic study of the bacterial microflora of 122 patients with gastric and duodenal ulcer was carried out. Microflora examination of the bioptic samples of mucosa, obtained from the ulcerous zone of the patients, revealed that an open ulcer is like an infected wound needing sanitation. In the focus of lesion microorganisms of 32 genera and species, including Helicobacter pylori, fungi of the genus Candida, representatives of the genera Streptococcus, Staphylococcus, Corynebacterium, Bacteroides, etc., were detected. Opportunistic microorganisms were isolated in associations (up to 8 different cultures), possessing cytotoxic, hemolytic, antilysozyme, lecithinase, caseinolytic and RNAase activities. To inhibit the microflora, chitosan was used; 82-85% of the cultures of different bacteria under study proved to be sensitive to it. The inclusion of chitosan into the complex therapy suppressed the persistence of H. pylori, ensured the sanitation of mucosa affected by opportunistic bacteria and accelerated ulcers cicatrization.     

Dysbioses of the oral cavity and intestines and immune reactivity in of adolescent bronchial asthma patients

Sixty bronchial asthma (BA) patients aged 15-18 years were examined between attacks. The composition of the oral cavity and intestine microflora as well as the total IgE level, peripheral blood eosinophils and lymphocytes expressing receptors CD3, CD4, CD8, CD22, CD11b, CD71, HLA-DR were detected. Colonization of the oral cavity by Staphylococcus aureus in 9%, by hemolytic streptococci in 10%, by fungi of the genus Candida in 25% and by enterococci in 7% of the patients was revealed. From the intestines S. aureus were isolated in 21%, hemolytic Escherichia coli in 18% and fungi of the genus Candida from 45% of the patients, while lactic acid bacteria were isolated only in 39% of the patients. The patients of this group were found to have eosinophilia (4.21 +/- 0.05%, p < 0.001) and a high concentration of IgE (330.2 +/- 29.31%, p < 0.01); in these patients the index CD4/CD8 was lower (< 0.05), the number of CD11b-, CD71- and HLA-DR positive lymphocytes was elevated. Thus, at periods between attacks the examined BA patients were found to have dysbiosis of mucous membranes, an elevated content of IgE, eosinophilia and lymphocyte activation indicative of the absence of immunological remission and required adequate diagnostic and therapeutic measures.     

Etiology of purulent septic diseases and the antibiotic resistance of the isolated causative agents

Among the causative agents of purulent septic diseases in the surgical hospital, 25 microbial species were isolated; of these, the prevailing species were Staphylococcus aureus (19.86 +/- 1.07%), Escherichia coli (16.5 +/- 0.99%) and Pseudomonas aeruginosa (10.06 +/- 0.8%). From environmental objects in the hospital 14 microbial species were isolated, among them bacteria of the genus Enterobacter (27 +/- 1.7%), E. coli (19.07 +/- 1.48%), S. aureus (14.7 +/- 1.31%), Klebsiella pneumoniae (13.73 +/- 1.31%), P. aeruginosa (7.33 +/- 0.98%). During 3 years of observation the isolation rate of K. pneumoniae from different environmental objects was found to increase threefold to 24.7 +/- 2.7%. The results of the study of the microbial picture in surgical hospitals, as well as the antibiotic resistance of circulating causative agents, should be borne in mind while taking epidemic control measures.     

Fatty and mycolic acid composition of Bacterionema matruchotii and related organisms.

Whole-organism methoanolysates of bacterionemae contained mycolic acids in addition to other long-chain fatty acids. These mycolic acids were similar in general structure and overall size to those found in strains of Corynebacterium diphtheriae and Corynebacterium xerosis. The long-chain fatty acids of bacterionemae, mainly straight-chain saturated and unsaturated acids, were similar to those of certain coryneform bacteria including C. diphtheriae. On the basis of these lipid data, and results of earlier studies, we recommend that the genus Bacterionema be transferred from the family Actinomycetaceae to the Coryneform Group of Bacteria.     

The relationship between the base composition of bacterial DNA and its intracellular melting temperature as determined by differential scanning calorimetry

The correlation between the melting temperature of intracellular DNA, determined by differential scanning calorimetry (DSC) of whole bacteria, and its guanine + cytosine (G + C) content, was examined for 58 species of bacteria. Samples of vegetative cells were heated in a Perkin-Elmer DSC-2C at 10 degrees C min-1 from 5 to 130 degrees C, cooled to 5 degrees C and then re-heated as before. Literature values for the mole fraction of G + C, XGC, were linearly related to the temperature, Tmax, at which the reversible peak, pr, observed on the second heating run was at a maximum, via the equation XGC = (Tmax -73.8)/41.0. This equation accounted for 91.9% of the variance in XGC with 95% confidence limits of +/- 7.3%, approximately 1.6 times the corresponding uncertainty (+/- 4.5%) quoted by De Ley (Journal of Bacteriology 101, 738-754, 1970) for estimates based on the spectroscopically determined melting temperature of purified DNA. Random errors of measurement of Tmax did not greatly limit the precision of the prediction and it was concluded that factors additional to base composition affected the temperature of DNA melting within the bacterial cell. Displacement of Tmax values from the fitted line was particularly noticeable in Campylobacter, Corynebacterium and Bacterionema species and part of the residual variation appeared to be species specific, possibly caused by differences in intracellular solute concentration.     

Isolation and Characterization of Haloanaerobacter chitinovorans gen. nov., sp. nov., a Halophilic, Anaerobic, Chitinolytic Bacterium from a Solar Saltern

Two halophilic anaerobic bacteria, one of which had chitinolytic activity, were isolated from a solar saltern in southern California. These organisms were long, gram-negative, motile, flexible rods. The biochemical and physiological characteristics of these bacteria were very similar but were different from the characteristics of other haloanaerobic bacteria. Both grew at salt concentrations ranging from 0.5 to 5 M and at temperatures ranging from 23 to 50 degrees C. They were sensitive to chloramphenicol but resistant to penicillin, carbenicillin, d-cycloserine, streptomycin, and tetracycline. An analysis of DNAs and whole-cell proteins showed that they were closely related taxonomically and distinguishable from other halophilic anaerobic bacteria. They exhibited 92.3 to 100% DNA homology as determined by DNA-DNA hybridization. The guanine-plus-cytosine contents of their DNAs were 34.8+/-1 mol%. The two isolates, strains W5C8 and W3C1, differed from other halophilic anaerobic bacteria sufficiently to support establishment of a new genus and species, Haloanaerobacter chitinovorans. Strain W5C8 exhibited chitinolytic activity and is designated the type strain. Two chitin-induced extracellular proteins with molecular weights of 38 x 10 and 40 x 10 were detected in strain W5C8.     

Corynebacterium aquatimens sp. nov., a lipophilic Corynebacterium isolated from blood cultures of a patient with bacteremia

An unknown lipophilic coryneform bacterium isolated from the blood cultures of a patient with bacteremia was characterized by phenotypic and molecular genetic methods. Chemical analysis revealed the presence of short chain mycolic acids consistent with the genus Corynebacterium. The DNA G+C content was 60.8mol%. Comparative 16S rRNA gene sequence analysis demonstrated that the isolate represents a new subline within the genus Corynebacterium. The closely phylogenetic relative of the unknown bacterium was found to be C. tuscaniense (97.8% sequence similarity). Partial rpoB gene sequence revealed that strain IMMIB L-2475(T) exhibited 13.5% sequence divergence with C. tuscaniense. The unknown bacterium was distinguished from C. tuscaniense by, DNA-DNA hybridization, cellular fatty acid profiles, MALDI-TOF analyses of cell extracts and biochemical tests. Based on the phylogenetic and phenotypic criteria, it is proposed that this bacterium be classified as new species, Corynebacterium aquatimens sp. nov., and is represented by strain IMMIB L-2475(T) (=DSM 45632(T)=CCUG 61574(T)).     

Identification of dimethyl disulfide-forming bacteria isolated from activated sludge

Twenty-four strains with high dimethyl disulfide (DMDS)-forming ability were isolated from activated sludge and identified to the genus level. These bacteria were classified into four groups (A, B, C, and D) by the API ZYM System (API System S.A., Montalieu, France). Group A (three strains) was identified as genus Lactobacillus by the API 20B System, by the method of Cowan and Steel, and by production of lactic acid as confirmed by gas-liquid chromatography. Group B (eight strains) was identified as genus Corynebacterium by API 20B and the Cowan and Steel method. Group C (one strain) was suggested to belong to genus Corynebacterium by the API 20B System. Group D (12 strains) was identified as genus Pseudomonas or Alcaligenes by the API 20B System, as genus Alcaligenes by the Cowan and Steel method, and as Achromobacter group Vd by the API 20NE System. However, on the basis of guanine-plus-cytosine contents in DNA and form of flagella, these strains were identified as genus Pseudomonas. Formation of DMDS from DL-methionine and S-methyl-L-cysteine was tested. DMDS-forming bacteria isolated from activated sludge formed DMDS from both precursors. In genus Pseudomonas, P. aeruginosa could not form DMDS from either precursor, but P. acidovorans, P. alcaligenes, P. pseudoalcaligenes, and P. testosteroni formed DMDS. In genus Alcaligenes, A. denitrificans subsp. xylosoxydans, A. denitrificans subsp. denitrificans, A. faecalis, and A. odorans formed DMDS from both precursors. Achromobacter group Vd formed DMDS from S-methyl-L-cysteine, but could not from DL-methionine.     

Comparison of the antibiotic sensitivity of lipophilic Corynebacterium sp. isolated from patients on the day of admission and during hospitalization

Lipophilic species of Corynebacterium are increasing problem in hospital infections. The aim of this study was to evaluate occurrence of these microorganisms in the materials taken from patients in the day of admission and during the hospitalization as well as comparison of their antibiotic sensitivity. The investigation included 65 strains isolated from hospitalized patients and 48 strains isolated from unchanged skin. Using Api Coryne test 5 species were identified. C. urealyticum dominated, the other were C. subsp. lipophilum and C. jeikeium. Among strains isolated from unchanged diseased skin the most C. jeikeium and C. accolens occurred. All strains were sensitive to glycopeptide, quinupristin/dalphopristin. The strains isolated from hospitalized patients were usually sensitive to fuside acid, doxycycline as well as tetracycline. Strains isolated from unchanged skin were sensitive to almost all tested antibiotics. In the group of 65 strains isolated from hospitalized patients 99.0% were multiresistant. In the group of strains isolated from unchanged skin only two strains were multiresistant. Differences in antibiotic sensitivity among analysed Corynebacterium sp. were confirmed. Majority of the "hospital strains" were characterized by multiresistance. Basing on these results it is possible to suppose, that multiresistance is main factor that favours lipophilic Corynebacterium species in the process of colonization of mucous membranes, skins as well as developing infections.