Setsuko Nishijima

Antimicrobial Agent of Susceptibilities and Antiseptic Resistance Gene Distribution among Methicillin-Resistant Staphylococcus aureus Isolates from Patients with Impetigo and Staphylococcal Scalded Skin Syndrome

ABSTRACT The susceptibilities to antimicrobial agents of and distributions of antiseptic resistance genes in methicillin-resistant Staphylococcus aureus (MRSA) strains isolated between 1999 and 2004 in Japan were examined. The data of MRSA strains that are causative agents of impetigo and staphylococcal scalded skin syndrome (SSSS) were compared with those of MRSA strains isolated from patients with other diseases. The susceptibilities to antiseptic agents in MRSA isolates from patients with impetigo and SSSS were higher than those in MRSA isolates from patients with other diseases. The distribution of the qacA/B genes in MRSA strains isolated from patients with impetigo and SSSS (1.3%, 1/76) was remarkably lower than that in MRSA strains isolated from patients with other diseases (45.9%, 95/207). Epidemiologic typings of staphylococcal cassette chromosome mec (SCCmec) and pulsed-field gel electrophoresis (PFGE) showed that MRSA strains isolated from patients with impetigo and SSSS had type IV SCCmec (75/76), except for one strain, and 64.5% (49/76) of the strains had different PFGE types. In addition, the patterns of restriction digestion of all tested qacA/B plasmid in MRSA isolates having different PFGE types were identical. The results showed that a specific MRSA clone carrying qacA/B was not prevalent, but qacA/B was spread among health care-associated MRSA strains. Therefore, it was concluded that the lower distribution rate of qacA/B resulted in higher susceptibilities to cationic antiseptic agents in MRSA isolated from patients with impetigo and SSSS.

The inhibition of free radical generation by human neutrophils through the synergistic effects of metronidazole with palmitoleic acid: a possible mechanism of action of metronidazole in rosacea and acne

SummaryMetronidazole is clinically effective in treating not only rosacea but also acne inflammation. Yet it is generally considered not to be very effective in inhibiting the growth of anaerobic Propionibacterium acnes. We report here our investigation into the synergistic effects of metronidazole and palmitoleic acid on the anaerobic growth of P. acnes as well as on human neutrophil functions, including the generation of reactive oxygen species (ROS). Both metronidazole and palmitoleic acid, when used alone, only slightly inhibited the growth of P. acnes, and no significant decrease in human neutrophil functions, including the generation of ROS, was observed. But metronidazole used in the presence of palmitoleic acid markedly inhibited the anaerobic growth of P. acnes and decreased ROS generation by neutrophils. However, ROS generated in the xanthine-xanthine oxidase system were not affected. Metronidazole was shown to be clinically effective by decreasing neutrophil-generated ROS at the sites of inflammation with the aid of palmitoleic acid, which is generally present in human skin. By inhibiting oxidative tissue injury under in vivo conditions, treatment with metronidazole results in remarkable improvement of rosacea and acne.

The bacteriology of acne vulgaris and antimicrobial susceptibility of Propionibacterium acnes and Staphylococcus epidermidis isolated from acne lesions.

We examined the species of bacteria aerobically and anaerobically isolated from 30 acne lesions and determined antimicrobial susceptibilities of Propionibacterium acnes (P. acnes) and Staphylococcus epidermidis (S. epidermidis) using nine antimicrobial agents. Among the bacteria isolated, S. epidermidis was most dominant. Both P. acnes and S. epidermidis were isolated from half of the acne lesions. The MIC of seven antimicrobials (ampicillin, erythromycin, roxithromycin, clindamycin, tetracycline, minocycline, nadifloxacin) against P. acnes was under 3.13 μg/ml. There were very few resistant strains of P. acnes, but many of S. epidermidis. More than 30% of the S. epidermidis isolates were resistant to erythromycin, roxithromycin, and clindamycin. After long‐term systemic antibiotic therapy, the resistant strains of S. epidermidis increased, but P. acnes resistance was still limited. When we use antimicrobial agents for the treatment of acne, it should be noticed that not only P. acnes but also S. epidermidis in the acne lesions may acquire resistance to antimicrobials.

Involvement of Propionibacterium acnes in the Augmentation of Lipogenesis in Hamster Sebaceous Glands In Vivo and In Vitro

Propionibacterium acnes is considered to be involved in the aggravation of acne vulgaris, but it remains unclear whether P. acnes directly influences lipogenesis in sebaceous glands. In this study, we showed that a culture medium of P. acnes (acnes-CM) and formalin-killed P. acnes (F-acnes) prepared from P. acnes strains, JCM6473 and JCM6425, intracellularly augmented lipid droplet formation and triacylglycerol (TG) synthesis in undifferentiated and insulin-differentiated hamster sebocytes. Acnes-CM and F-acnes prepared from four clinical P. acnes strains elicited the same lipogenesis augmentation. The augmented TG production resulted from an increase in the diacylglycerol acyltransferase activity. Topical application of acnes-CM to the skin of hamster auricles every day for 4 weeks revealed that sebum accumulation was augmented in sebaceous glands and ducts. Furthermore, both acnes-CM and F-acnes increased the production of 15-deoxy-Delta(12,14)-prostaglandin J(2) (15d-PGJ(2)), a cytochrome P450 (CYP)-linked sebaceous lipogenic factor, in differentiated sebocytes. A CYP inhibitor, SKF-525A, decreased the acnes-CM- and F-acnes-augmented production of TG and 15d-PGJ(2). Thus, to our knowledge these results provide previously unreported evidence that P. acnes directly participates in the augmentation of sebaceous lipogenesis through a proposed mechanism in which an increase of 15d-PGJ(2) production through the CYP pathway is closely associated with the enhancement of TG production.

Antimicrobial susceptibilities of Propionibacterium acnes isolated from patients with acne vulgaris.

Antibiotic susceptibilities of Propionibacterium acnes in Japan were determined. Erythromycin‐resistance was found in 10.4% (5/48) of the strains, and four of these were cross‐resistance to clindamycin. Although the erythromycin ribosome methylase gene erm(X) was looked for, no strain carrying erm(X) was found. Sequencing analysis revealed that all of the erythromycin‐resistant strains had a mutation in the peptidyl transferase region of the 23S rRNA gene: G2057A, A2058G, or A2059G. Consequently, our results show that P. acnes resistance to macrolides is caused by a mutation in the 23S rRNA gene, and has been increasing in Japan.

The Incidence of Isolation of Methicillin-resistant Staphylococcus aureus (MRSA) Strains from Skin Infections during the Past Three Years (1989–1991)

We did a statistical study of 294 strains of Staphylococcus aureus (S. aureus) isolated from skin infections during the period from January of 1989 to December of 1991 in the Department of Dermatology, Kansai Medical University Hospital. We especially examined methicillin‐resistant S. aureus (MRSA) from the point of view of incidence, variety of skin infections with MRSA, coagulase type, phase type, and resistance against antimicrobial agents. The frequency of isolation of MRSA has been increasing. In 1991, the proportion of MRSA isolates among all S. aureus strains isolated from skin infections was 41.5%. MRSA was isolated most often from infectious decubitus. Coagulase type II and phage group NT (not typable) MRSA were most frequently isolated. The resistance of MRSA to OFLX and IMP/CS had remarkably increased. Notably, the resistance to MINO was low before 1991.

Antimicrobial resistance of Staphylococcus aureus isolated from skin infections.

The antimicrobial susceptibility of 229 strains of Staphylococcus aureus isolated from various skin infections was determined against 22 antimicrobial agents by the agar dilution method. The clinical isolates were most sensitive to vancomycin, teicoplanin, mupirocin and fusidic acid. No strains were resistant to vancomycin or teicoplanin. Three strains were highly resistant (MIC > or =100 mg/l) to mupirocin and eight strains to fusidic acid. The MIC(50) of all antimicrobials, except for gentamicin, were below 3.13 mg/l. The incidence of resistance to penicillin, cephalosporins and clindamycin ranged from 20 to 30%. The occurrence of gentamicin, erythromycin and roxithromycin resistance was high at 55.2, 39.6 and 39.1%, respectively. Methicillin resistance occurred in 21.0% of strains. The incidence of organisms with MIC > or =3.13 mg/l to oxacillin was 24.3%. These results were comparable to the average rate of MRSA in Japanese dermatological specimens.

Clinical and bacteriologic evaluation of OPC-7251 in patients with acne: A double-blind group comparison study versus cream base

Twenty-eight patients with acne were assigned to 4 weeks of treatment with OPC-7251 (a new fluoroquinolone derivative) 1% cream or the cream base in a double-blind manner to evaluate the antibacterial effect of the drug on resident bacteria in the hair follicles and to evaluate clinical response. Propionibacterium acnes was isolated from 21 of the 28 acne patients. When the number of P. acnes was compared before and after treatment, the posttreatment P. acnes count in the OPC-7251 1% cream group was significantly (p = 0.000) reduced compared with that in the cream base group. OPC-7251 1% cream was also significantly (p = 0.019) superior to the cream base in terms of clinical response. P. acnes and Staphylococcus epidermidis isolated from the acne lesions were selected for their susceptibility to various antibacterial agents. The minimal inhibitory concentration of OPC-7251 against P. acnes and S. epidermidis was 0.10 to 0.20 and 0.024 to 0.10 micrograms/ml, respectively, which indicates that the drug has a potent antibacterial effect.

The antibiotic susceptibility of Propionibacterium acnes: a 15-year bacteriological study and retrospective evaluation.

Propionibacterium acnes strains were isolated from the comedones of 46 acne patients, and their susceptibilities to penicillin G, ampicillin, erythromycin, clindamycin, tetracycline, doxycycline, minocycline, cephalexin and gentamycin were studied by examining their minimum inhibitory concentrations. Two additional standard strains were included in the study. In the results, differing antibiotic susceptibilities were found, and P. acnes strains were the most sensitive to erythromycin and clindamycin, followed by ampicillin and minocycline. The highest levels of resistance were observed against clindamycin, erythromycin and tetracycline. Non‐resistant strains were detected only against minocycline. A comparison of these results with others obtained by similar studies in our department during the last 15 years showed a progressive tendency toward antibiotic resistance for Propionibacterium acnes, probably related to the systemic administration of antibiotics for the treatment of bacterial infections other than acne.

Malassezia folliculitis is caused by cutaneous resident Malassezia species

Malassezia folliculitis [MF] is caused by the invasion of hair follicles by large numbers of Malassezia cells, but it remains unclear which Malassezia species are involved in the disease. To clarify this situation, Malassezia species isolated from lesions of MF patients were analyzed by both culture and non-culture methods. In addition, Malassezia species recovered from the non-lesion areas of the skin of MF patients and skin samples of healthy subjects were included in this study. The test population consisted of 32 MF patients and 40 healthy individuals. The lesions were obtained using a comedone extractor, while swabs were employed to obtain skin samples from non-lesion areas of the patients and healthy subjects. Malassezia DNA was analyzed using a real-time PCR technique. The detection limit of the culture method was 5 CFU/cm(2) as opposes 50 cells/cm(2) with non-culture procedures. The predominant species recovered from MF lesions were M. globosa and M. sympodialis by culture method analysis, and M. restricta, M. globosa, and M. sympodialis with non-culture methods. These results were in agreement with those found with samples from non-lesion skin areas of MF patients and healthy subjects. This study clarified that MF is caused by Malassezia species that are part of the cutaneous microflora and not by exogenous species.