[Development and Evaluation of Screening Culture Medium for Detection of Drug-Resistant Gram Negative Rods Containing Stealth Type CPE].

There is a report that an infection by medicine resistant bacteria will be the number one cause of death in 2050 according to the recommendation of WHO, and the CPE (carbapenem-producing Enterobacteriaceae) infection is regarded as a problem in particular. When detecting CPE, it is important how to detect stealth type CPE sensitive to carbapenem series medicines. So we used the 2 types of screening culture medium, "KBM" CRE-JU culture medium (CRE-JU culture medium) and the FRPM culture medium, and tried to detect drug-resistant gram-negative bacilli such as CPE, stealth type CPE, ESBL-producing bacteria, and excess AmpC-producing bacteria (AmpC-producing bacteria), etc. in combination of this culture mediums. As a result, CRE-JU culture medium showed a difference in the growth of CPE depending on the amount of inoculated bacteria while ß-lactamase non-producing strain and other strains except for high concentration ESBL-producing bacteria and AmpC-producing bacteria were un-growing. Most of the CRE, stealth type CPE, ESBL-producing bacteria and AmpC-producing bacteria grew in the FRPM culture medium while most of the ß-lactamase non-producing strains with a MIC value of meropenem (MEPM) of 2 µg/mL or less were un-growing. From these results, it was suggested that when a strain grown on CRE-JU and FRPM culture mediums, it could be distinguished as CPE, and when strains grown on FRPM culture medium which were un-grown on CRE-JU culture medium, it could be distinguished as drug-resistant bacteria such as stealth type CPE, ESBL-producing bacteria, and AmpC-producing bacteria. When strains not grown on CRE-JU and FRPM culture mediums, it could be distinguished as sensitive.

Correlations between Resistance Classifications Based on Penicillin-Binding Protein Genotypes and Antimicrobial Susceptibility Test Results of Haemophilus influenzae.

There are several problems associated with antimicrobial susceptibility testing (AST) of Haemophilus influenzae. ß-Lactamase-negative ampicillin-resistant H. influenzae (BLNAR) isolates with minimum inhibitory concentration (MIC) of ampicillin (ABPC) <4 mg/l will be classified as susceptible according to the MIC breakpoint of the CLSI M100 criteria, in spite of harboring penicillin-binding protein (PBP) mutations that cause ABPC resistance. A total of 103 isolates were collected from clinical materials for analysis. The genotypes of the PBP mutations were analyzed by polymerase chain reaction. The WalkAway 96 Plus (WALKAWAY), dry plate Eiken (DP-EIKEN), and RAISUS S4 systems (RAISUS) were used for AST. HTM broth was used as the culture medium for WALKAWAY, Mueller‒Hinton broth with 5% lysed horse blood for DP-EIKEN, and HTM with 5% horse serum for RAISUS. The MIC concordance rates of ABPC for g-BLNAR, for RAISUS vs. DP-EIKEN, RAISUS vs. WALKAWAY, and DP-EIKEN vs. WALKAWAY were 96.1, 86.4, and 85.4%, respectively. WALKAWAY had a low correlation with the other two systems. Moreover, concordance rates of ABPC MIC ≥4 mg/l, which is considered as resistant, of 69 g-BLNAR isolates for the RAISUS, DP-EIKEN, and WALKAWAY systems were 68.1, 58.0, and 37.7%, respectively. Therefore, in Japan, where the BLNAR strain is isolated at a high frequency, it is necessary to understand the characteristics of the measuring systems to appropriately interpret the test results.

[Investigation of Skin Adherent Bacterial Flora in Dogs by MALDI-TOF MS and 16S rRNA Gene Analysis].

Due to the increase in the number of companion animal breeders in Japan, there are more opportunities for companion animals to come into contact with humans than before. Therefore, we investigated the bacterial flora adhering to the skin of dogs and the bacterial flora was analyzed for the presence of zoonotic bacteria that infect humans from companion animals. With the cooperation of students enrolled in the Department of Medical Technology and Science, Faculty of Fukuoka Health Care, International University of Health and Welfare. 39 samples were collected from the abdomen, back and paws of 13 healthy dogs using sterile swabs by the scraping method. The isolation culture was carried out only for facultative anaerobic bacteria to obligate aerobic bacteria and Bacterial identification was determined by MALDI-TOF MS and 16S rRNA gene analysis. Among the identified strains were Pasteurella canis, Staphylococcus pseudintermedius, Staphylococcus intermedius, which were difficult to detect in humans. The overall ratio of detected bacteria was 35% for coagulasenegative staphylococci, 14% for coagulase-positive staphylococci, 5% for Enterobacteriaceae, and 45% for natural environment. In the future, it is expected that extended-spectrum ß-lactamase producing bacteria and drug-resistant bacteria such as Carbapenem-resistant enterobacterales will also be transmitted to humans through contact with companion animals.