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Corynebacterium mooreparkense, a later heterotypic synonym of Corynebacterium variabile.

Strains of a Gram-positive bacterium were isolated from the Irish smear-ripened cheese Gubbeen, and assigned to a new species, Corynebacterium mooreparkense, in 2001. During a further study on the same cheese, no additional isolates from this species could be found. Instead, multiple isolates of its nearest phylogenetic neighbour, Corynebacterium variabile, were found. A first screening with rep-PCR and SDS-PAGE pointed to a similarity between C. mooreparkense and C. variabile. Following this peculiar result, attempts were made to collect all type strains deposited at different culture collections and all strains described by Brennan et al. [Int J Syst Evol Microbiol (2001) 51, 843-852]. Subsequently, 16S rRNA gene sequencing and DNA-DNA hybridizations were performed. All C. mooreparkense strains had a 16S rRNA gene sequence similarity of at least 99.5 % with C. variabile and the DNA-DNA relatedness was 95 %. On the basis of these results, it is concluded that C. mooreparkense is a later heterotypic synonym of C. variabile.

Bacterial Proteins↗

Susceptibilities of Corynebacterium bovis and Corynebacterium amylocolatum isolates from bovine mammary glands to 15 antimicrobial agents.

Coryneform bacteria are frequently isolated from bovine mastitis and are associated with economic losses. Generally, the MICs of the 15 antimicrobial agents tested at which 90% of the isolates tested are inhibited for 46 Corynebacterium bovis and 13 Corynebacterium amylocolatum strains were low. These are the first quantitative antimicrobial susceptibility data available for coryneforms from bovine mastitis. Data from this study suggest that comparable corynebacteria from humans have a much higher level of antimicrobial resistance to a variety of antimicrobial agents.

Animals↗

Cloning of the pyruvate kinase gene (pyk) of Corynebacterium glutamicum and site-specific inactivation of pyk in a lysine-producing Corynebacterium lactofermentum strain.

The pyruvate kinase gene pyk from Corynebacterium glutamicum was cloned by applying a combination of PCR, site-specific mutagenesis, and complementation. A 126-bp DNA fragment central to the C. glutamicum pyk gene was amplified from genomic DNA by PCR with degenerate oligonucleotides as primers. The cloned DNA fragment was used to inactivate the pyk gene in C. glutamicum by marker rescue mutagenesis via homologous recombination. The C. glutamicum pyk mutant obtained was unable to grow on minimal medium containing ribose as the sole carbon source. Complementation of this phenotype by a gene library resulted in the isolation of a 2.8-kb PstI-BamHI genomic DNA fragment harboring the C. glutamicum pyk gene. Multiple copies of plasmid-borne pyk caused a 20-fold increase of pyruvate kinase activity in C. glutamicum cell extracts. By using large internal fragments of the cloned C. glutamicum gene, pyk mutant derivatives of the lysine production strain Corynebacterium lactofermentum 21799 were generated by marker rescue mutagenesis. As determined in shake flask fermentations, lysine production in pyk mutants was 40% lower than that in the pyk+ parent strain, indicating that pyruvate kinase is essential for high-level lysine production. This finding questions an earlier hypothesis postulating that redirection of carbon flow at the phosphoenol pyruvate branch point of glycolysis through elimination of pyruvate kinase activity results in an increase of lysine production in C. glutamicum and its close relatives.

Amino Acid Sequence↗

Identification of Turicella otitidis isolated from a patient with otorrhea associated with surgery: differentiation from Corynebacterium afermentans and Corynebacterium auris.

Turicella otitidis is a newly described coryneform bacterium isolated from middle ear fluids. We report here on the diagnosis of a strain isolated from otorrhea. The API Coryne system (bioMérieux, Marcy I'Etoile, France) used alone failed to differentiate T. otitidis, Corynebacterium afermentans, and Corynebacterium auris (ANF group). Biochemical tests such as DNase, enzymatic reactions (API ZYM; bioMérieux), and carbon substrate assimilation tests (Biotype 100; bioMérieux) allow presumptive identification. However, only chemotaxonomy and molecular biology can achieve unequivocal differentiation among these three species.

Cerebrospinal Fluid Otorrhea↗

Comparative study of Corynebacterium parvum and Corynebacterium liquefaciens on antitumor activity against sarcoma-180.

Corynebacterium parvum and Corynebacterium liquefaciens were comparatively examined for their antitumor activity against sarcoma-180 in ddY mice. In the case of ascitic form, significant antitumor effect was observed when C. parvum was administered on days -4 and -2 or day -2. As for solid form, maximal effect was obtained when C. parvum was administered on day 0. On the other hand, C. liquefaciens exhibited maximal antitumor activity against sarcoma-180, both in ascitic and solid forms, when it was administered on days -4 and -2. However, no significant difference in respect to antitumor activity against sarcoma-180 was seen between C. parvum and C. liquefaciens (ascitic form, F1 118 = 0.09, P greater than 0.05; solid form, F1 53 = 0.03, P greater than 0.05).

Animals↗

Detection of differences in the nucleotide and amino acid sequences of diphtheria toxin from Corynebacterium diphtheriae and Corynebacterium ulcerans causing extrapharyngeal infections.

While Corynebacterium ulcerans can mimic classical diphtheria, extrapharyngeal infections are extremely rare. Sequencing of the diphtheria toxin (DT)-encoding tox gene of two C. ulcerans isolates from extrapharyngeal infections revealed differences from C. diphtheriae DT sequences, mainly in the translocation and receptor-binding domains. C. ulcerans supernatants were much less potent than supernatant from C. diphtheriae. A C. ulcerans DT-specific PCR is described below.

Adult↗

Immunomodulation of host resistance to experimental viral infections in mice: effects of Corynebacterium acnes, Corynebacterium parvum, and Bacille calmette-guérin.

Resistance to a representative group of experimental virual infections in mice was significantly enhanced by nonspecific modulation of host defense mechanisms. Corynebacterium acnes, Corynebacterium parvum, and bacille Calmette-Guérin were effective in enhancing host resistance. Animals treated seven to 10 days before inoculation of virus were protected against a lethal infection with Herpesvirus hominis type 2, encephalomyocarditis virus, murine cytomegalovirus, or Semliki Forest virus. The protection of experimental animals against encephalomyocarditis virus infection intitiated by either the intraperitoneal or the respiratory route indicated that C. acnes exerted a systemic, rather than local, effect. A maturation process was required for host defense mechanisms stimulated by C. acnes, as indicated by the failure to enhance resistance in suckling animals. Involvement of cells of the lymphoreticular system was demonstrated by transfer of enhanced resistance against H. hominis type 2 to recipient animals with peritoneal exudate cells harvested from mice pretreated with C. acnes. Finally, these same cells inhibited the progression of herpetic infection in tissue culture. The data suggest that immunomodulation, possibly through activation of macrophages, may offer a method for enhancement of host resistance to viral infections.

Animals↗

Adhesion of Corynebacterium renale and Corynebacterium pilosum to the epithelial cells of various parts of the bovine urinary tract from the renal pelvis to vulva.

The various parts of the bovine urinary tract, the renal pelvis, the ureter, the urinary bladder, the urethra, the vaginal vestibule and the vulva, were examined for the capacity of the epithelial cells to bind Corynebacterium renale and C. pilosum. C. renale adhered best to the epithelial cells of the vulva, and then to those of the ureter and renal pelvis. C. pilosum also adhered best to the epithelial cells of the vulva, followed by those of the vaginal vestibule. The results indicate that the most important target tissue for these bacteria may be the vulva, and the results correlate with the fact that C. renale frequently causes pyelonephritis and ureteritis, while C. pilosum causes the same diseases less frequently and behaves like normal flora of the vaginal vestibule.

Adhesiveness↗

Most Corynebacterium xerosis strains identified in the routine clinical laboratory correspond to Corynebacterium amycolatum.

A comprehensive study was performed on 25 bacterial clinical isolates originally identified as Corynebacterium xerosis. Three reference strains of C. xerosis were also included in the study. On the basis of a variety of phenotypic characteristics tested, all strains could be divided into two separate clusters: reference strains ATCC 373 (the type strain of C. xerosis) and ATCC 7711 showed yellow-pigmented, dry, rough colonies, fermented 5-keto-gluconate, exhibited strong leucine arylamidase and alpha-glucosidase activities, produced lactate as the major end product of glucose metabolism, were susceptible to most of the 19 antimicrobial agents tested, and showed an inhibition zone around disks containing the vibriocidal compound O/129. In contrast, the remaining 26 strains including reference strain NCTC 7243 as well as all clinical isolates formed white-grayish, dry, slightly rough colonies, did not ferment 5-keto-gluconate, exhibited only weak leucine arylamidase and no alpha-glucosidase activity, produced large amounts of propionic acid as the end product of glucose metabolism, and were resistant to most antimicrobial agents tested, including O/129. Chemotaxonomic (cellular fatty acids, mycolic acids, and G+C content) and molecular genetic (16S rRNA gene sequence) investigations revealed that the strains of the second cluster unambiguously belonged to the species C. amycolatum. Our data suggest that most strains reported in the literature as C. xerosis are probably misidentified and correspond to C. amycolatum.

Bacterial Typing Techniques↗

Evaluation of differential media for the identification of Corynebacterium genitalium and Corynebacterium pseudogenitalium (group JK corynebacteria).

Tween purple agar containing 1% fructose (TFP agar) differentiated Corynebacterium genitalium from C. pseudogenitalium, which respectively formed colorless and yellow colonies after 72 h incubation at 37 degrees C aerobically or in 5-10% CO2 in air. Thus TFP agar is a differential medium. Corynebacteria-like colonies grown on nonspecific urethritis (NSU) chocolate agar from urogenital material were identified as C. genitalium, C. pseudogenitalium, or commensals when subcultured on TPF agar. TFP agar was unsuitable for their primary isolation since the commensals turned the medium yellow with 24 h incubation. Gentamicin cannot be employed as a selective agent in medium for the isolation of these corynebacteria. TFP agar containing 10 micrograms/mL entamicin inhibited most strains of C. pseudogenitalium and C. genitalium isolated from urogenital infections. It did not inhibit isolates of these corynebacteria from cancer patients or suppress the normal bacterial flora of the urogenital tract. Evidence that gentamicin-resistant strains are characteristic of nosocomial infections is presented.

Corynebacterium↗

Corynebacterium CDC group JK (Corynebacterium jeikeium) sepsis in haematological patients: a report of three cases and a systematic literature review.

We describe 3 patients with Corynebacterium jeikeium sepsis in neutropenic phase during treatment for acute myeloid leukaemia. Fever was the first symptom. All had a central venous catheter which was removed. Two patients developed subcutaneous nodules containing pus when the neutrophil count recovered; 1 had intracutaneous and pulmonary lesions. They were treated with vancomycin and recovered when the neutrophil count started to rise. A review of 80 neutropenic patients with C. jeikeium sepsis reported in the literature, together with our 3 cases indicates that risk factors for infection are the presence of a central venous catheter, being an adult male or postmenopausal female, profound and prolonged neutropenia and exposure to multiple antibiotics. Skin lesions are reported in 48% and pulmonary lesions in 36% of the patients. The overall mortality is 34% but in patients with recovery of the bone marrow only 5%. Therefore haematopoietic growth factors should be considered in neutropenic patients with C. jeikeium infection.

Adult↗

Efficient electrotransformation of corynebacterium diphtheriae with a mini-replicon derived from the Corynebacterium glutamicum plasmid pGA1.

Efficient transformation of the human pathogen Corynebacterium diphtheriae was achieved with novel cloning vectors consisting of a mini-replicon from the cryptic C. glutamicum plasmid pGA1 as well as of the aph(3')-IIa or tetA(Z ) antibiotic resistance genes. Plasmid-containing transformants of C. diphtheriae were recovered at frequencies ranging from 1.3 x 10(5) to 4.8 x 10(6) colony forming units (cfu)/microg of plasmid DNA. Vector DNA was directly transferred from Escherichia coli into C. diphtheriae with frequencies up to 5.6 x 10(5) cfu/microg of plasmid DNA. On the basis of the pGA1 mini-replicon, an expression vector system was established for C. diphtheriae by means of the P(tac) promoter and the green fluorescent reporter protein. In addition, other commonly used vector systems from C. glutamicum, including the pBL1 and pHM1519 replicons, and the sacB conditionally lethal selection marker from Bacillus subtilis, were shown to be functional in C. diphtheriae. Thus, the ability to apply the standard methods of C. glutamicum recombinant DNA technology will greatly facilitate the functional analysis of the recently completed C. diphtheriae genome sequence.

Biotechnology↗

Comparison of surface hydrophobicity of piliated and non-piliated clones of Corynebacterium renale and Corynebacterium pilosum.

Piliated (P+) and non-piliated (P-) clones of Corynebacterium renale and C. pilosum were similar in hydrophobicity as measured by hydrophobic interaction chromatography, bacterial adherence to hydrocarbons and the salt aggregation test. Therefore, the previously reported adherence of P+ clone to various cells, which is more effective than that of P- clone, may be uncorrelated with the degree of hydrophobicity of both clones of these bacteria. Hydrophobicity of P+ and P- clones was found to be high when measured by hydrophobic interaction chromatography and bacterial adherence to hydrocarbons, but low when measured by the salt aggregation test.

Bacterial Adhesion↗

Isolation and characterization of monoclonal antibodies against pili of Corynebacterium renale and Corynebacterium pilosum.

Five monoclonal antibodies against pili of Corynebacterium renale 115 P+ (piliated clone) and two monoclonal antibodies against pili of C. pilosum 92 P+ (piliated clone) were produced. These antibodies bound to pili of the homologous strain in in enzyme-linked immunosorbent assay (ELISA) and agglutinated P+ but not P- (non-piliated clone) of each homologous strain. The five monoclonal antibodies against C. renale 115 P+ pili were divided into 2 groups, comprising 16/5, 160/1 and 32/6 and 13/4 and B20/3, based on the results of a competitive binding assay. The results may indicate the presence of at least 2 distinct antigenic areas on the pilus of C. renale 115 P+. The monoclonal antibodies of the first group inhibited adhesion of C. renale 115 P+ bacteria to the epithelial cells of bovine vulva, while the second group did not. Two monoclonal antibodies against C. pilosum 92 P+ pili recognized the same area on the pilus of C. pilosum 92 P+, and inhibited the adhesion of C. pilosum 92 P+ bacteria to the epithelial cells of bovine vulva. The adhesion of these bacteria was inhibited by the monoclonal antibodies in the form of IgG as well as by the Fab fragment. The strains of C. renale and C. pilosum which reacted with each of the anti-C. renale 115 P+ pili and anti-C. pilosum 92 P+ pili monoclonal antibodies were small in number and of restricted distribution.

Agglutination Tests↗

Heterogeneity within Corynebacterium minutissimum strains is explained by misidentified Corynebacterium amycolatum strains.

Forty-eight clinical strains that were tentatively identified as Corynebacterium minutissimum on the basis of standard biochemical reactions (Hollis-Weaver tables) as well as by the use of the API (RAPID) Coryne system were examined further. Two different groups of strains were observed. The first group (including the type strain of C minutissimum) contained 27 strains showing creamy colonies. These strains grew homogeneously in 6.5% NaCl broth, exhibited DNase activity, were susceptible to the vibriocidal compound O/129, produced succinic acid, and contained mycolic acids. The second group comprised 21 strains with dry colonies. They grew in clumps at the surface of 6.5% NaCl broth, DNase activity was not detected, they were resistant against O/129, produced large amounts of propionic acid, and mycolic acids were not detected. In combination with quantitative DNA-DNA hybridizations, it was demonstrated that strains of the second cluster belonged, in fact, to C amycolatum. Furthermore, it was observed that a few C minutissimum strains may also ferment mannitol. These data indicate that the clinical microbiologist must be careful not to misidentify C amycolatum strains as C minutissimum.

Bacterial Infections↗

In-vitro activity of psychiatric drugs against Corynebacterium urealyticum (Corynebacterium group D2).

We tested the in-vitro activity of amoxycillin, amoxycillin/clavulanic acid, cefotaxime, gentamicin, trimethoprim-sulphamethoxazole, tetracycline, norfloxacin, ciprofloxacin, vancomycin, teicoplanin, clindamycin and five psychiatric drugs (chlorpromazine, sertraline, fluoxetine, paroxetine and risperidone) against 32 strains of Corynebacterium urealyticum. Resistance rates exceeded 90% for all antibiotics except glycopeptides, quinolones and tetracycline. Sertraline was the most active psychiatric drug. We tested the influence of sertraline on the activity of amoxycillin, amoxycillin/clavulanic acid, cefotaxime, gentamicin, trimethoprim-sulphamethoxazole, tetracycline and ciprofloxacin. We did not observe antagonism in any case. Sertraline enhanced the activity of ciprofloxacin and tetracycline against all strains (MIC decrease: 4-64-fold for ciprofloxacin, 2-32-fold for tetracycline).

1-Naphthylamine↗

Relationship between group JK corynebacteria and the biotypes of Corynebacterium genitalium and Corynebacterium pseudogenitalium.

Twenty-six strains of group JK corynebacteria had the same colonial morphology and biological reactions as the biotypes of the biovars of Corynebacterium genitalium and C. pseudogenitalium. Therefore, group JK corynebacteria can be assigned to the biovars of C. genitalium or C. pseudogenitalium. Although the strains differed in sensitivity to 16 antibiotics tested by Sensi-Discs or by the Micro-Media technique, they are uniformly sensitive to 4-5 micrograms/mL of vancomycin. Medium containing 10 micrograms vancomycin/mL was bactericidal and the killing time was dependent on the concentration. The rate of mutation to resistance to 10 micrograms vancomycin was greater than 1 in 10(10) corynebacteria. Therefore, vancomycin sensitivity is a stable characteristic of these corynebacteria which also indicates that group JK corynebacteria are strains of either C. genitalium or C. pseudogenitalium. Since group JK corynebacteria are considered pathogens, this finding supports the belief that C. genitalium is a pathogen and suggests that some biotypes of the commensal C. pseudogenitalium may infect compromised hosts.

Anti-Bacterial Agents↗