Evaluation of hydrocortisone as a strain-dependent growth-regulator of Porphyromonasgingivalis.

OBJECTIVE: Porphyromonas gingivalis is an oral key pathogen and known to be very diverse in geno- and phenotypes. It is a fastidious bacterium with low O2-tolerance and 3-7 days of incubation are necessary. With growing interest in the field of microbial endocrinology we explored the potential growth-stimulating effect of hydrocortisone (HC, synonym cortisol) on P. gingivalis cultures. MATERIAL AND METHODS: Six different P. gingivalis strains were pre-incubated in supplemented Brain-Heart-Infusion broth under appropriate conditions for 24 h, diluted and transferred into microplates. A newly developed and semi-automated spectrophotometric measurement in triplicate, applying a SpectraMax i3x microplate reader at an optical density of 600 nm, was conducted to test growth differences between test group (exposed to a supplement of either 1.25, 2.5, 5, 10, or 20 µg/ml of hydrocortisone) and control group over 48 h of anaerobic incubation (O2 ≤ 1%). Furthermore, strains were also incubated on HC-supplemented blood agar to test for a possible growth-stimulating effect on solid media. RESULTS: HC significantly stimulated the lag-phase growth of four out of six P. gingivalis strains. Our data suggest a concentration-dependent growth stimulatory effect of HC between 2.5 and 5 µg/ml, while below 1.25 µg/ml and above 10 µg/ml HC either did not stimulate or inhibited growth. CONCLUSIONS: HC could reduce the incubation time when isolating P. gingivalis from clinical samples and could boost low biomass cultivations especially during their lag-phase. The growth-modulating effect might be via modulation of virulence factors/quorum sensing gene expression or by reactive oxygen species(ROS)-capturing during early stages of bacterial growth. Further experiments are necessary to explain the mechanism behind our observations.

The Antimicrobial Susceptibility of Porphyromonas gingivalis: Genetic Repertoire, Global Phenotype, and Review of the Literature.

The in vitro antimicrobial susceptibility of 29 strains of the major periodontal pathogen Porphyromonas gingivalis and three P. gulae (as an ancestor) to nine antibiotics (amoxicillin, amoxicillin/clavulanate, clindamycin, metronidazole, moxifloxacin, doxycycline, azithromycin, imipenem, and cefoxitin) was evaluated by E-testing of minimal inhibitory concentration (MIC) according to international standards. The results were compared with 16 international studies reporting MICs from 1993 until recently. In addition, 77 currently available P. gingivalis genomes were screened for antimicrobial resistance genes. E-testing revealed a 100% sensitivity of P. gingivalis and P. gulae to all antibiotics. This was independent of the isolation year (1970 until 2021) or region, including rural areas in Indonesia and Africa. Regarding studies worldwide (675 strains), several method varieties regarding medium, McFarland inoculation standards (0.5-2) and incubation time (48-168 h) were used for MIC-testing. Overall, no resistances have been reported for amoxicillin + clavulanate, cefoxitin, and imipenem. Few strains showed intermediate susceptibility or resistance to amoxicillin and metronidazole, with the latter needing both confirmation and attention. The only antibiotics which might fail in the treatment of P. gingivalis-associated mixed anaerobic infections are clindamycin, macrolides, and tetracyclines, corresponding to the resistance genes erm(B), erm(F), and tet(Q) detected in our study here, as well as fluoroquinolones. Periodical antibiotic susceptibility testing is necessary to determine the efficacy of antimicrobial agents and to optimize antibiotic stewardship.