Hello everybody!
Does anybody know the optimal culture conditions of Propionibacterium acnes?
Someone has just isolated phages against P.acnes?
Thank you in advance for your answers!

hi
P acnes is a pleomorphic Gram (+) bacillus which is normally found in and around hair follicles. The organism is anaerobic but can tolerate limited amounts of oxygen (microaerophilic).
P.acnes is a difficult organism to culture due to the slow growth time and the anaerobic conditions required for optimum growth. The generation doubling time for P.acnes is 5.1 hours

Mar:

My facility has had success with growing P acnes on spread plates of Reinforced Clostridial Agar (RCA) incubated anaerobically at 37C for 5 - 7 days. Note that the colonies are very small.

Please reply with a post if you have discovered additional culture condition information.

m a masters of pharmacy student. i need proponibacterium acne culture..how do i prepare it?

amit pharm1:

Hoeffler, et al (1976) evaluated the antimicrobial susceptibility of 38 strains of P. acnes. "For primary isolation, agar containing
0.43 M glycerol was used. Further cultivation was done on A-agar medium, the contents of which have been described by Jong et al. Bacterial suspensions were prepared in Mueller-Hinton broth (Difco), and the antimicrobial drugs tested were added to Mueller-Hinton medium (Difco, pH 7.2)."

"Preparation of inocula - Cells from 48-h-old cultures
on A-agar medium were added to Mueller-Hinton broth, carefully shaken to establish a homogeneous suspension."

References:

U. Hoeffler, H. L. Ko, AND G. Pulverer. Antimicrobial Susceptibility of Propionibacterium acnes and Related Microbial Species. ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, Sept. 1976, Vol. 10, No. 3 p. 387-394

Jong, E. C., H. L. Ko, and G. Pulverer. 1975. Studies on bacteriophages of Propionibacterium acnes. Med. Microbiol. Immunol. 161:263-271.

DENYS, et al (1983) evaluated a total of 104 P. acnes isolates recovered from "clinical specimens in diverse geographical
areas and sent between 1973 and 1982 to the CDC as single isolates for identification."

"Organisms to be tested were grown on anaerobic blood agar
plates (Nolan Biological Labs, Inc., Atlanta,Ga.) at 35°C for 48 to 72 h in an anaerobic chamber (Forma Scientific, Marietta, Ohio)."


References:

GERALD A. DENYS, ROBERT C. JERRIS, JANA M. SWENSON, AND CLYDE THORNSBERRY. Susceptibility of Propionibacterium acnes Clinical Isolates to 22 Antimicrobial Agents. ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, Feb. 1983, p. 335-337 Vol. 23, No. 2

Fuimura and Nakamura (1978) used P. acnes CN-8 which produces acnecin, and a laboratory stock
culture strain P. acnes EXC-1.

"Cultivation was carried out at 37°C anaerobically in an anerobic glove box (COY Manufacturing Co., Ann Arbor, Mich.) for 4 days in a medium containing 3.7% brain heart infusion (Difco Laboratories, Detroit, Mich.) supplemented with 0.2% yeast extract (Difco)."


Reference:

SETSUO FUJIMURA AND TAKESHI NAKAMURA
Purification and Properties of a Bacteriocin-Like Substance (Acnecin) of Oral Propionibacterium acnes. ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, Dec. 1978, Vol. 14, No. 6 p. 893-898

Tyrell, et al (2006) evaluated the "the in vitro activities of daptomycin, vancomycin, and penicillin against more than 100 strains each of Clostridium difficile, C. perfringens, Finegoldia magna, and Propionibacterium acnes".

"The isolates were obtained from human clinical specimens, including blood, skin and skin structure, intraabdominal, and feces, identified by standard methods (Jousimies-Somer, 2002) and maintained in skim milk at -70°C. The strains were taken from the freezer and transferred at least twice on brucella agar (Anaerobe Systems, Morgan Hill, CA) supplemented with hemin, vitamin K1, and 5% sheep blood (brucella blood agar [BBA]) to ensure purity and good growth."

"Time-kill studies were performed in reduced brucella broth supplemented with vitamin K1 and hemin."



References:

Kerin L. Tyrrell, Diane M. Citron, Yumi A. Warren, Helen T. Fernandez, C. Vreni Merriam, and Ellie J. C. Goldstein. In Vitro Activities of Daptomycin, Vancomycin, and Penicillin against Clostridium difficile, C. perfringens, Finegoldia magna, and Propionibacterium acnes. ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, Aug. 2006, p. 27282731 Vol. 50, No. 8

Jousimies-Somer, H. R., P. Summanen, D. M. Citron, E. J. Baron, H. M. Wexler, and S. M. Finegold. 2002. Wadsworth-KTL anaerobic bacteriology manual. Star Publishing, Belmont, Calif.

Wang et al (1977) "examined the in vitro susceptibility of 96 strains ofP. acnes to 17 antibiotics"

"Antimicrobial susceptibility was done by using the agar dilution technique on brain heart infusion agar supplemented with hemin (5 ,ug/ml) and vitamin K (0.5 ,ug/ml). Stock solutions of the test drugs were prepared in sterile distilled water; further dilutions were made in brain heart infusion broth supplemented with
hemin (5 ,g/ml) and vitamin K1 (1.0 jig/ml) and added to similarly supplemented brain heart infusion agar at 46°C to yield final drug concentrations ranging from 0.1 to 50 jig/ml."

"The inoculum was prepared by diluting a 24-h supplemented brain heart infusion broth culture to provide an inoculum size of approximately 104 colony-forming units"

All plates were incubated in an anaerobic chamber at 36°C for 48 hr.



References:

WEN LAN LOU WANG, E. DALE EVERETT, MARCIA JOHNSON, and EVELYN DEAN . Susceptibility of Propionibacterium acnes to Seventeen Antibiotics Antimcrobial. AGENTS AND CHEMOTHERAPY, Jan. 1977, Vol. 11, No. 1 P. 171-173

Goldstein et al (2006) evaluated strains which were "isolated from clinical skin specimens obtained between 1984 and 2003 from adult patients and identified by standard criteria (Jousimies-Somer,, 2002) and stored in skim milk at -70°C. Frozen cultures were subcultured twice on brucella agar supplemented with hemin, vitamin K1, and 5% sheep blood (Anaerobe Systems, Morgan Hill, CA) to ensure purity and good growth."

"Susceptibility testing was performed according to CLSI (formerlyNCCLS) standards (NCCLS document M11-A6), with
brucella agar supplemented with hemin, vitamin K1, and 5%
laked sheep blood and an inoculum of 10^5 CFU per spot"


References:

Ellie J. C. Goldstein, Diane M. Citron, C. Vreni Merriam, Yumi A. Warren, Kerin L. Tyrrell, and Helen T. Fernandez. Comparative In Vitro Activities of Retapamulin (SB-275833) against141 Clinical Isolates of Propionibacterium spp., Including 117 P. acnes Isolates. ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, Jan. 2006, Vol. 50, No. 1 p. 379381

Jousimies-Somer, H. R., P. Summanen, D. M. Citron, E. J. Baron, H. M. Wexler, and S. M. Finegold. 2002. Wadsworth-KTL anaerobic bacteriology manual, 6th ed. Star Publishing Co., Belmont, Calif.