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DNA sequence homology between attB-related sites of Corynebacterium diphtheriae, Corynebacterium ulcerans, Corynebacterium glutamicum, and the attP site of gamma-corynephage.

Chromosomal restriction fragments of Corynebacterium ulcerans and C. diphtheriae, containing an integration site for corynephages of the beta family, show homology on Southern blots. Homologous DNA in also found in the soil isolate C. glutamicum, although this strain is not susceptible to beta-corynephages. Three of these DNA fragments, one for each bacterial strain, and a fragment of gamma-corynephage DNA previously shown to contain the phage integration site, were cloned and sequenced. Alignment of the 3 bacterial sequences shows a very high degree of homology in a stretch of ca 120 nucleotides, whereas the rest of the sequences is generally non-homologous. Within this common bacterial portion, a segment of ca. 96 nucleotides (core sequence) is also highly homologous to the phage sequence. The first half (ca. 50 bp) of the core sequence is identical in all aligned sequences whereas the second half, which is largely occupied by a stem-and-loop structure, contains point mutations peculiar to each clone. The described sequences are likely to be involved in phage integration/excision processes.

Attachment Sites, Microbiological

In vitro susceptibility of Corynebacterium group D2 and Corynebacterium jeikeium to twelve antibiotics.

The in vitro susceptibility of 83 Corynebacterium group D2 strains and 44 Corynebacterium jeikeium strains to 12 antimicrobial agents was determined by an agar dilution technique using Mueller-Hinton agar supplemented with Tween 80 (0.025%). All strains of Corynebacterium group D2 were highly sensitive to fusidic acid, pristinamycin, teicoplanin and vancomycin (geometric mean MICs 0.047, 0.048, 0.338 and 0.396 mg/l respectively). Most of the strains were resistant to other antibiotics tested (ciprofloxacin, erythromycin, gentamicin, lincomycin, rifampin and tetracycline). However, a few strains were highly sensitive (MICs less than or equal to 0.2 mg/l). The overall pattern of susceptibility of 44 strains of Corynebacterium jeikeium was similar; the geometric mean MICs of fusidic acid, pristinamycin, vancomycin and teicoplanin were 0.234, 0.235, 0.557 and 0.652 mg/l respectively.

Anti-Bacterial Agents

Corynebacterium group D2 ("Corynebacterium urealyticum") constitutes a new genomic species.

Twenty-one Corynebacterium group D2 ("C. urealyticum") strains were found to constitute a tight DNA hybridization group distinct from named Corynebacterium species. The strains of Corynebacterium group D2 had cell wall component type IV, short chain mycolic acids and G+C content of DNA of 65-66 mol %. Corynebacterium group D2 constitutes a genomic species which can be identified by phenotypic tests.

Base Composition

[The Corynebacterium pyogenes infection of cattle. 2. Tenacity of Corynebacterium pyogenes].

Some common agents were tested for their effectiveness against Corynebacterium pyogenes. The pathogen proved most susceptable to Wofasteril. All germs were killed within ten minutes by a 0.005% solution. Equally good action was recorded from all the other tested agents as well (lactic acid, Lugol's solution, formalin, cupric sulphate, alcohol, and aethacridine. Other studies were conducted with the view to testing the survival capacity of Corynebacterium pyogenes in different media and storage conditions. The pathogen survived three months in routine media and mastitis secretion at room temperature. Regrowth of 38 in 50 strains took place after nine months of refrigerator storage in slanting blood agar tubes with paraffin plugs. Germs sampled from mastitis secretion and stored in a refrigerator were cultivable even after one year had elapsed. The detectability rate of Corynebacterium pyogenes did not change over months by storage of wound infection material at 12 degrees C below zero. The pathogen remained detectable five days from artificial contamination of cattle skin.

Animals

Multiresistant corynebacteria in bacteriuria: a comparative study of the role of Corynebacterium group D2 and Corynebacterium jeikeium.

During an 11-month prospective study, urine cultures were performed on 5685 samples obtained in three hospital units. The use of a selective medium improved the recovery of antibiotic-multiresistant corynebacteria (AMC): 703 isolates (12.4%) compared with 88 isolates (1.6%) on sheep blood agar. Corynebacterium group D2 (CGD2) was isolated in 80.5% of urines yielding greater than or equal to 10(5) AMC ml-1 whereas Corynebacterium jeikeium represented 80.2% of isolates with less than 10(5) AMC ml-1. Among 16 patients with greater than or equal to 10(5) ml-1 C. jeikeium none had signs of urinary tract infection. In contrast, among 56 patients with greater than or equal to 10(5) CGD2, 40 (71%) had abnormal urinary sediment (mainly apatite or struvite crystals) and 29 (52%) had clinical signs of urinary tract infections sometimes complicated by lithiasis (seven cases) and alkaline-encrusted cystitis (two cases).

Anti-Infective Agents, Urinary

The specificity of enzymes adding amino acids in the synthesis of the peptidoglycan precursors of Corynebacterium poinsettiae and Corynebacterium insidiosum.

A soluble extract from Corynebacterium poinsettiae able to synthesize the nucleotide precursor of ite peptidoglycan was prepared. This extract contained all the enzymes necessary for the synthesis of the peptide side-chain. The spedificity of these enzymes was determined and compared with the specificity of similar enzymes extracted from the closely related Corynebacterium insidiosum. In both organsims, addition of the third amino acid of the peptide side-chain was specific for the amino acid and nucleotide dipeptide involved in peptidoglycan synthesis in the parent organism. L-Diaminobutyric acid, which is found as the acetyl derivative in the precursor nucleotide and in the completed peptidoglycan of C. insidiosum, was added as the free amino acid and not as the acetylated compound.

Alanine

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

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

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

[Effect of Corynebacterium parvum on the immune response in guinea pigs. I. Mode of enhancement of the anamnestic response and development of delayted hypersensitivity after treatment with Corynebacterium parvum].

The effect of Corynebacterium parvum (C. parvuum) on the immune response of the guinea pig to ovalbumin varies with the protocol of immunization. The marked effect of C. parvum on the anamnestic response in the rabbit has been confirmed in the guinea pig when immunization is carried out intradermally with a mixture of C. parvum and ovalbumin. When C. parvum is given intravenously or subcutaneously or intradermally but separately from the antigen, this effect is not observed. Whatever the route of injection guinea pigs treated with C. parvum show skin reactions of delayed type hypersensitivity at the site of an intradermal booster when the latter is given at least 27 days after primary immunization.

Animals

[Effect of Corynebacterium parvum on the immune response in guinea pigs. II. Passive transfer of enhanced anamnestic response and delayed hypersensitivity observed after treatment with Corynebacterium parvum].

After intradermal immunization with a mixture of Corynebacterium parvum (C. parvum) and ovalbumin guinea pigs show a markedly increased anamnestic response to an intradermal booster of ovalbumin as compared to controls treated with ovalbumin only. At the same time a reaction of delayed type hypersensitivity is observed in the treated animals, but not in controls. The enhanced anamnestic response as well as the posivitive skin reaction were transferred to strain 2 histocompatible guinea pigs by peripheral blood leukocytes as well as by peritoneal exudate cells. Passive transfer was not obtained after prior irradiation of donor animals.

Animals

[Activity of ethambutol, isoniazid and rifampicin on Corynebacterium urealyticum and Corynebacterium jeikeium].

Because corynebacteria and mycobacteria have walls of similar composition, it was of interest to test the effects on resistant corynebacteria of two agents with anti-wall effects, i.e., ethambutol (EMB) and isoniazid (INH). Rifampicin was also studied to extend previous data and clarify conflicting results in the literature. INH was not active in levels achieved in vivo with standard dosages. The minimal inhibitory concentration (MIC) of EMB was under serum levels for only three of 78 strains. All minimal bactericidal concentration (MBCs) were greater than serum levels. With RIF, 59% of strains were susceptible and 19.2% exhibited intermediate susceptibility as defined by the French Committee on Antimicrobial Susceptibility Testing. CMI and CMB distributions were heterogeneous, suggesting a possible acquired heterogeneous resistance to RIF.

Corynebacterium

[Development of the vector-host system in Corynebacterium. Cloning and expression of homoserine dehydrogenase and homoserine kinase genes in Corynebacterium cells].

Novel cloning vectors for glutamic acid producing bacteria have been constructed. The cryptic plasmid pBO1 (4.4 kb) from Brevibacterium sp. recombined with the plasmid pACYC184 (4.0 kb) from Escherichia coli was used to produce composite plasmid named pKA1. The plasmid could propagate and express the Cm-r phenotype in E. coli and coryneform glutamic acid producing bacteria Br. flavum, C. glutamicum, Br. lactofermentum. The pKA1 plasmid and its variants deleted within non-essential plasmid regions with unique restriction sites HindIII, SalGI, SphI were used in cloning experiments. The genes coding for threonine biosynthesis of C. glutamicum and Br. flavum were subcloned into shuttle vectors in C. glutamicum cells. Recombinant plasmids were introduced into protoplasts by polyethylenglycol-mediated transformation of plasmid DNAs. It was shown that the presence of plasmids containing the Br. flavum thrA2 gene in C. glutamicum (thrB) caused 10-fold increase in homoserine dehydrogenase activity, as compared to that of wild type strain, and in homoserine production.

Alcohol Oxidoreductases