Prevalence of antimicrobial-resistant Cutibacterium isolates and development of multiplex PCR method for Cutibacterium species identification.

INTRODUCTION: Cutibacterium species such as C. acnes, C. avidum, and C. granulosum are known anaerobic skin inhabitants and often cause surgical site infections. These species are genetically similar and are difficult to identify rapidly. In addition, their pathogenicity and antimicrobial resistance remain unknown. In this study, antimicrobial resistance in Cutibacterium isolates was studied and a multiplex PCR method for their identification was developed. METHODS: A total of 497 C. acnes, 71 C. avidum, and 25 C. granulosum strains which were isolated from the acne pustule and infectious regions, were used. RESULTS: The antimicrobial resistance rates of C. acnes, C. avidum, and C. granulosum strains isolated from patients with acne vulgaris were higher than those of strains isolated from patients with infectious diseases. In particular, macrolide-clindamycin-resistant strains were isolated most frequently from all species. Among the resistant strains, strains with 23S rRNA mutations were the most common in C. acnes (24.3%, 71/292), whereas C. avidum and C. granulosum strains were most frequently found with erm(X). For the first time, a C. granulosum strain carrying pTZC1, which codes erm(50) and tet(W), was isolated from patients with acne vulgaris. Regarding the rapid identification of causative pathogens from infectious regions, three Cutibacterium species were identified with 100% sensitivity and specificity using multiplex PCR method. CONCLUSIONS: Our data showed that antimicrobial resistance differed among Cutibacterium species. The multiplex PCR method may contribute to the rapid detection of Cutibacterium species and selection of appropriate antimicrobial agents.

Cutibacterium acnes phylogenetic type IC and II isolated from patients with non-acne diseases exhibit high-level biofilm formation.

Cutibacterium (formerly Propionibacterium) acnes is an important for not only exacerbating factor of acne vulgaris but also pathogen of surgical site infections (SSIs) in orthopedics and plastic surgery. Although biofilm-forming (BF) C. acnes are associated with intractable SSI, characteristics of these strains were still unknown. Here, we explored detailed molecular epidemiological features of BF C. acnes isolated as causative pathogen of infectious diseases. Phylogenetic types of 205 C. acnes strains isolated between 2013 and 2018 from 18 clinical departments of a university hospital in Japan were determined by single-locus sequence type (SLST). Clade H (traditional type IC) and K (type II) which are less relevant with healthy skin and acne vulgaris, were detected in 26.8% (55/205) and 16.1% (33/205) of the strains, respectively. The incidence of them was significantly higher than that of acne patients (H and K, each 2.9%, P < 0.05). In addition, SLST distribution of C. acnes strains differed by each department and isolation site. When biofilm formation was quantified, 51 strains (24.9%) were defined as high-BF strains. Notably, most high-BF strains were classified into the strains of clade H (56.4%, 31/55) and clade K (54.4%, 18/33), and these strains were frequently found in the strains isolated from patients of medical emergency center and plastic surgery. Similarly, high-BF strains were frequently found among the isolates from blood (35.7%) and catheters (30.0%), with a high proportion belonging to clades H and K. Compared to C. acnes strains isolated from acne patients, antimicrobial-resistant strains were less identified in non-acne patients. Our findings showed that pathogenicity of C. acnes strains differs by their phylogenetic types. Furthermore, we showed clade H and K have the ability of high biofilm formation and suggest that these strains have potential to become a risk factor for SSI.

An outbreak of severe infectious diseases caused by methicillin-resistant Staphylococcus aureus USA300 clone among hospitalized patients and nursing staff in a tertiary care university hospital.

The USA300 clone, which produces Panton-Valentine leukocidin (PVL), is a major highly pathogenic community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA) clone that is spreading throughout the world. Although the prevalence of the USA300 clone in Japan was very limited a decade ago, its incidence has been increasing in both community and hospital settings in recent years. There is great concern that the USA300 clone will cause more complicated diseases and become a serious threat to immunocompromised patients in hospital settings. Here, we report an outbreak of severe infectious diseases in a tertiary care university hospital involving the incidence of deep infections, including bacteremia, and continuous and frequent isolation of MRSA strains for five months from six patients and a healthy nursing staff member in the same ward. The genotype of all MRSA isolates was identical to that of the USA300 clone. Furthermore, pulsed-field gel electrophoresis analysis indicated that all MRSA had the same patterns. These data demonstrate that a USA300 clone outbreak had occurred in the hospital. Fortunately, this outbreak was terminated subsequent to the interventions of the infection control team and all patients recovered following the appropriate therapies. Our report demonstrates that patients carrying highly pathogenic CA-MRSA have the potential to become a source of nosocomial outbreaks that can spread to healthy healthcare workers. Therefore, stricter standard precautions should be applied for all patients at the time of admission to prevent such nosocomial outbreaks.

Group A Streptococcus pharyngitis outbreak among university students in a judo club.

We report on an outbreak of Group A Streptococcus (GAS) pharyngitis among university students in a judo club. Eventually, 14 of total 23 club members developed acute pharyngitis clinically. In a span of 15 days in April 2013, 12 students visited our hospital complaining of sore throat and high fever. All were men with a median age of 19.5 years (interquartile range, 18-21). The rapid streptococcal antigen test was positive in 3 of 4 patients (75%) without previous antibiotic treatment, and in 2 of 8 patients (25%) with previous antibiotic treatment. The definitive diagnosis of GAS pharyngitis was made by either a positive RADT or positive throat culture of GAS when patients had more than 2 findings from the Centor scoring system in this study. 5 students received the definitive diagnosis. The throat culture results showed that 1 out of 9 asymptomatic students was GAS-positive. The outbreak might have occurred by person-to-person contact while living in a dormitory and during judo training, which is a highly close-contact sport. However, there was also the possibility of oral transmission by the shared use of water bottles, although the culture from 1 bottle was negative. Some students continued to participate in the judo club activities after the onset of sore throat or fever. Healthcare professionals, teachers, and coaches should be aware of the potential outbreaks of infectious diseases among university students engaged in athletic activities. Furthermore, it is important to educate athletes about infectious diseases.

Emergence of multidrug-resistant Pandoraea sputorum in Japan.

Pandoraea is a pathogenic bacterium naturally resistant to various antimicrobials, including colistin. Here, we report the whole-genome sequence of Pandoraea sputorum, which exhibits high-level multidrug resistance, isolated from a hospitalized patient in Japan.

Multidrug-resistant Cutibacterium avidum isolated from patients with acne vulgaris and other infections.

OBJECTIVES: Cutibacterium avidum, a human skin commensal bacterium, rarely causes infections. It has recently been shown that Cutibacterium acnes, another member of the genus, acts as an opportunistic pathogen in surgical site infections. However, the antimicrobial susceptibility and pathogenicity of C. avidum remain unknown. METHODS: We investigated the epidemiological features and antimicrobial susceptibility of C. avidum isolated from patients with acne vulgaris and other infections. RESULTS: Cutibacterium avidum strains were isolated from patients with acne vulgaris (29 strains) and other infections (12 strains). Clarithromycin and clindamycin resistance was observed in 65.9% (27/41) of strains. In addition, ciprofloxacin resistance was observed in 34.1% (14/41) of strains, of which 13 also exhibited resistance to macrolides and clindamycin. Notably, the macrolide-clindamycin resistance gene erm(X) was found on the chromosome of 92.6% (25/27) of clindamycin-resistant strains and may be prevalent owing to transmission among C. avidum strains. Ciprofloxacin-resistant strains developed amino acid substitutions in GyrA owing to the use of antimicrobial agents. Pulsed-field gel electrophoresis (PFGE) analysis revealed that only a few strains exhibited 100% similarity. Additionally, no clustering associated with antimicrobial resistance, biofilm-forming ability or type of infection was observed. CONCLUSION: Our study revealed that erm(X) may be frequently disseminated in C. avidum, and multidrug-resistant C. avidum strains may colonise the skin of patients with acne vulgaris and other infections. Therefore, the prevalence of multidrug-resistant C. avidum and the use of antimicrobial agents for the treatment of acne vulgaris and other infections associated with C. avidum should be monitored.

Antimicrobial susceptibilities and distribution of resistance genes for beta-lactams and macrolides in Streptococcus pneumoniae isolated between 2002 and 2004 in Tokyo.

Antimicrobial susceptibilities of 205 Streptococcus pneumoniae strains isolated between 2002 and 2004 in Japan were examined and the distribution of genes for resistance to penicillins and macrolides were investigated by polymerase chain reaction. The molecular epidemiology of 92 randomly selected isolates was also examined by pulsed-field gel electrophoresis (PFGE). The numbers of S. pneumoniae isolates resistant to benzylpenicillin, clarithromycin and tetracycline were, respectively, 39 (19%), 111 (54%) and 155 (76%), and the numbers increased annually. All isolates were susceptible to amoxicillin, fluoroquinolones, vancomycin and linezolid. Analysis of mutations in the genes for penicillin-binding protein showed that 92% of isolates had mutations in pbp1a, pbp2b and/or pbp2x. Susceptibility to benzylpenicillin decreased with increasing number of mutated pbp genes. The macrolide resistance genes ermB and mefA were found in 99 (48%) and 76 (37%) isolates, respectively. The presence of ermB was associated with high-level resistance to macrolides, and the percentage of isolates with ermB increased annually. The presence of mefA also increased with increasing number of mutated pbp genes. Although the 92 isolates belonged to 74 PFGE types, three groups with an 80% similarity in their PFGE patterns were found at high frequency. Two of the three groups contained no isolates susceptible to penicillin and/or tetracycline, and their percentages increased annually. Our results suggest that the number of S. pneumoniae isolates with reduced susceptibility due to accumulation of resistance genes has been increasing.

Using the tannase gene to rapidly and simply identify Staphylococcus lugdunensis.

The coagulase-negative Staphylococcus lugdunensis, a bacterium similar to Staphylococcus aureus, produces tannase that degrades tannin. We developed a polymerase chain reaction-based method to rapidly and simply identify this species by detecting the tanA gene for S. lugdunensis tannase.