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Filamentous phage active on the gram-positive bacterium Propionibacterium freudenreichii.

We present the first description of a single-stranded DNA filamentous phage able to replicate in a gram-positive bacterium. Phage B5 infects Propionibacterium freudenreichii and has a genome consisting of 5,806 bases coding for 10 putative open reading frames. The organization of the genome is very similar to the organization of the genomes of filamentous phages active on gram-negative bacteria. The putative coat protein exhibits homology with the coat proteins of phages PH75 and Pf3 active on Thermus thermophilus and Pseudomonas aeruginosa, respectively. B5 is, therefore, evolutionarily related to the filamentous phages active on gram-negative bacteria.

Amino Acid Sequence↗

Studies on bacteriophages of Propionibacterium acnes.

With the help of adaptation experiments, 61 phage preparations out of 36 Propionibacterium acnes bacteriophages (32 isolated by us and 4 sent from abroad) were established. On the basis of their stability, spectrum of activity, and virulence, 13 phages were selected for phagetyping. 58 well-classified P. acnes strains were grouped into 7 phage-types. 7 strains of P. granulosum, two strains of P. avidum, and one yet ungroupable microserophilic propionibacterium strain were resistant to all 61 phages, even when tested in 100 X RTD. A combination of phagetyping with biotyping resulted in data especially useful for the differentiation of P. acnes.

Bacteriophage Typing↗

Characterization of the novel Cutibacterium acnes phage KIT08 and its associated pseudolysogenic bacterial isolate.

Cutibacterium acnes, formerly Propionibacterium acnes, is a Gram-positive bacterium commonly recognized as an important factor in acne vulgaris and infections associated with prosthetic medical devices. With the rise in antibiotic resistance, phage therapy has gained renewed attention as a promising alternative to antibiotics. In addition to a strict lytic cycle, some virulent phages may enter a pseudolysogenic state and exclude superinfections, thereby significantly limiting the applicability of these potential antimicrobial agents. However, the trade-off induced by phage infection of bacterial cells during this state and its molecular mechanism are yet to be confirmed, especially for C. acnes phages. In this study, a novel Cutibacterium acnes phage, KIT08, was isolated and characterized. It demonstrated rapid infectivity and moderately strong bacteriolysis. After infection of C. acnes NBRC 107,605, pseudolysogenic bacteria were collected and examined for physiological tradeoffs. The pseudolysogenic isolate exhibited slower growth and downregulation of the transcriptional levels of biofilm-producing genes, such as lipase 2 and hyaluronate lyase, leading to a decrease in biofilm formation. Additionally, a genomic study of phage KIT08 revealed that open reading frames 23 and 34 encode putative proteins homologous to repressor C and LTP proteins, which may play an important role in the induction of pseudolysogeny and superinfection exclusion in C. acnes.

Propionibacterium acnes↗

Use of bacteriophage typing to distinguish Propionibacterium acne types I and II.

Strains of serotypes I and II of Propionibacterium were compared for phage sensitivity. The two serotypes could be distinguished by using a typing set consisting of 16 bacteriophages at concentrations that demonstrated selective lysis of serotype I or II bacterial strains. Seven phage types were found; three were composed exclusively of serotype I, and four were exclusively composed of serotype II organisms. Generally, serotype I strains were more sensitive to phage lysis than were serotype II strains. No correlation was found between phage type and site of isolation.

Bacterial Infections↗

Properties of Corynebacterium acnes bacteriophage and description of an interference phenomenon.

Nine virulent bacteriophages of the anaerobe Corynebacterium (Propionibacterium) acnes, the P-a series, are DNA phages, with long, curved nonretractile tails (130 nm) without tail plates or fibers. They have isometric heads (420 by 460 nm), and are placed in Bradley's group B-1. There is permanent plaque suppression at highest phage concentrations. After 100- to 1,000-fold dilution, plaques are evident. The latent period is 1 h and burst size 25. Cross-neutralization data of antisera for the nine phages are similar. There is an unexplained precipitous drop in plaque-forming units during the first 5 min of neutralization, after which the rate is linear for 2 h. They are sensitive to pH extremes but are partially protected even at pH 4 or 9 by storage at 4 C. They are resistant to ether and chloroform and are inactivated within 10 min at 70 C.

Antigens, Bacterial↗

Propionibacteria in patients with acne vulgaris and in healthy persons.

A total of 375 anaerobic and microaerophilic coryneform rods, isolated from the pilosebaceous ducts of 26 healthy persons (71 strains) and from comedones (93 strains), pustules (107 strains), and the unaffected skin (104 strains) of 36 acne patients were classified according to the species key in Bergey's manual, the biotyping scheme of Pulverer and ko, the serotyping schedule of Höffer et al., and the phage typing schedule of Jong et al. The statistical evaluation demonstrated certain differences in the frequencies of the Propionibacterium species and types between the different groups tested. Thus, the species P. granulosum was isolated only from acne patients (50.0% of patients examined) and was found more frequently in comedones and pustules than in unaffected follicles in acne patients. The majority of P. granulosum strains belonged to serotype II (95). Biotype A propionibacteria were more frequently found in strains from healthy controls (52.1% of strains) than in strains from healthy controls (52.1% of strains) than in strains from comedones (17.2%), pustules (27.1%) and unaffected skin (38.5%) of the acne patients. The results of phage-typing showed that the P. acnes lysotype I was more frequent in acne patients (total: 73.2% of strains), especially in the inflamed pustules (88.5%), than in healthy controls (55.1%).

Acne Vulgaris↗

Adenosylcobalamin-dependent methylmalonyl-CoA mutase from Propionibacterium shermanii. Active holoenzyme produced from Escherichia coli.

The linked structural genes coding for both subunits of adenosylcobalamin-dependent methylmalonyl-CoA mutase from the Gram-positive bacterium Propionibacterium shermanii have been altered by site-directed mutagenesis and placed under the control of an inducible phage-T7-specific plasmid promoter in Escherichia coli. Conditions have been found under which both alpha- and beta-subunits are produced in soluble form, in near 1:1 ratio, and assemble to form apo-mutase totalling about 5% of the total cellular protein. Methylmalonyl-CoA mutase purified from these cells could be readily converted into the holoenzyme by addition of adenosylcobalamin. The active holoenzyme apparently crystallizes in the same space group as an inactive corrinoid-containing form of the enzyme obtained previously.

Base Sequence↗

Immunotherapy and cytokines.

Immunomodulation is now possible in veterinary medicine with the licensure of a number of biological products by the United States Department of Agriculture for veterinary use. These products activate primarily macrophages, induce the production of cytokines, and have various effects on the activity and proliferation of B and T lymphocytes. Those products most commonly used are inactivated whole bacteria of Propionibacterium acnes, cell wall fractions of nonpathogenic Mycobacterium spp, and the lysate derived from lysis of Staphylococcus aureus by a bacterial phage. All products have been licensed for use against specific diseases, but the literature includes studies for off-label usage. These immunomodulators are considered to be nonspecific stimulators of the immune system and may affect both humoral and cellular functions of immunity.

Adjuvants, Immunologic↗

Qualitative and quantitative investigations on the resident bacterial skin flora in healthy persons and in the non-affected skin of patients with seborrheic Eczema.

The composition of the aerobic and anaerobic bacterial skin flora was investigated quantitatively and qualitatively in 25 patients with seborrheic eczema (SE) and in 35 healthy persons. In SE patients, the geometric mean count for propionibacteria in the pilosebaceous ducts of the forehead was reduced to a statistically significant extent in comparison with the control group. There was the same tendency with regard to the geometric mean count of the superficial anaerobic bacterial flora of the back, whereas the geometric mean counts of the coagulase-negative staphylococci were the same in the two groups. These results may explain earlier findings of a reduction of free fatty acids in the surface lipids in SE patients. According to the results of differentiation in 295 propionibacteria and 157 micrococcaceae, 7 biotypes and 10 phage types of propionibacteria which were so far unknown could be demonstrated. Significant differences between the groups with regard to the frequency of different biotypes, serotypes, and phage types could be demonstrated neither in propionibacteria nor in coagulase-negative staphylococci. The results of typing, however, showed that patients with SE displayed much more frequently several different propionibacterial species and types as well as different micrococcal species and types than persons with healthy skin.

Adult↗

Differentiation of two groups of Corynebacterium acnes.

One hundred and forty-three strains of Corynebacterium acnes, isolated from human skin and acne lesions, were compared with three strains of Propionibacterium acnes from the American Type Culture Collection. The 146 organisms could be separated into two groups. Members of the larger group (129 strains) hydrolyzed gelatin and usually produced indole, Gel-In(+), but were unable to ferment trehalose, maltose, or sucrose, TMS(-). The deoxyribonucleic acid from selected strains of this group had an average guanosine + cytosine (GC) content of 60.5%. The members of the smaller group (17) were Gel-In(-), TMS(+), and the deoxyribonucleic acid had an average GC content of 63.9%. Studies with absorbed and unabsorbed antisera to the smaller group showed that although there were antigens shared by the two groups, it was possible to distinguish them serologically. Members of each group produced propionic acid. The principal fatty acid component of members of each group was iso-C(15) fatty acid. Seventy per cent of the Gel-In(+) strains were lysed by phage 174, whereas only one of 15 Gel-In(-) strains was lysed. Pending further information on the genetics of the two groups, those Gel-In(-), TMS(+) strains are tentatively designated C. acnes, group II.

Agar↗

Alpha-melanocyte-stimulating hormone reduces endotoxin-induced liver inflammation.

Alpha-Melanocyte-stimulating hormone (MSH) is a potent anti-inflammatory agent in many models of inflammation, suggesting that it inhibits a critical step common to different forms of inflammation. We showed previously that alpha-MSH inhibits nitric oxide (NO) production in cultured macro-phages. To determine how alpha-MSH acts in vivo, we induced acute hepatic inflammation by administering endotoxin (LPS) to mice pretreated with Corynebacterium parvum, alpha-MSH prevented liver inflammation even when given 30 min after LPS administration. To determine the mechanisms of action of alpha-MSH, we tested its influence on NO, infiltrating inflammatory cells, cytokines, and chemokines. Alpha-MSH inhibited systemic NO production, hepatic neutrophil infiltration, and increased hepatic mRNA abundance for TNF alpha, and the neutrophil and monocyte chemokines (KC/IL-8 and MCP-1). We conclude that alpha-MSH prevents LPS-induced hepatic inflammation by inhibiting production of chemoattractant chemokines which then modulate infiltration of inflammatory cells. Thus, alpha-MSH has an effect very early in the inflammatory cascade.

Animals↗

Antibody coated bacteria in otitis media with effusions.

The significance of bacterial isolates of coagulase negative Staphylococcus epidermidis and Corynebacterium species in middle ear fluids remains controversial. This study was undertaken to evaluate the possible role of these organisms in different effusions. Cultures were obtained from the external cartilaginous ear, bony canal, tympanic membrane, middle ear fluids, tonsils and nasopharynx of 93 patients at the time of myringotomy for otitis media with effusion. Isolates of coagulase negative staphylococci from the middle ear, external ear and posterior aural skin were evaluated in regard to antibiotic sensitivity, biochemical tests and phage typing. Furthermore, in another series of patients in whom these organisms were isolated, specific antibody activity in both middle ear washings and corresponding sera were measured, using the technique of indirect immunofluorescence. The results suggest that the isolates from the middle ear and ear canal are not always identical. There is some evidence that specific IgM and IgG as well as IgA responses to these organisms are present in middle ear washings and not in corresponding serum. These results suggest that coagulase negative Staphylococcus and Corynebacterium species in middle ear effusions may not be contaminants from the external canal and that a local immune response may be produced by these organisms. Whether or not these organisms represent etiologic agents responsible for otitis media with effusion cannot be ascertained from this data. One very interesting point is the presence of IgF coating of organisms in some effusions.

Adolescent↗