METHODS: CLSI broth microdilution methodology was used to determine antimicrobial activity and EUCAST breakpoints version 9.0 were used to determine rates of susceptibility and resistance. Isolates were also screened for the genes encoding extended-spectrum β-lactamases (ESBLs) or carbapenemases (including metallo-β-lactamases [MBLs]).

RESULTS: Between 2015 and 2017, this study collected a total of 7051 Enterobacterales isolates and 2032 Pseudomonas aeruginosa isolates from hospitalized patients in Australia, Japan, South Korea, Malaysia, the Philippines, Taiwan, and Thailand. In the Asia-Pacific region, Enterobacterales isolates that were ESBL-positive, carbapenemase-negative (17.9%) were more frequently identified than isolates that were carbapenemase-positive, MBL-negative (0.7%) or carbapenemase-positive, MBL-positive (1.7%). Multidrug-resistant (MDR) isolates of P. aeruginosa were more commonly identified (23.4%) than isolates that were ESBL-positive, carbapenemase-negative (0.4%), or carbapenemase-positive, MBL-negative (0.3%), or carbapenemase-positive, MBL-positive (3.7%). More than 90% of all Enterobacterales isolates, including the ESBL-positive, carbapenemase-negative subset and the carbapenemase-positive, MBL-negative subset, were susceptible to amikacin and ceftazidime-avibactam. Among the carbapenemase-positive, MBL-positive subset of Enterobacterales, susceptibility to the majority of agents was reduced, with the exception of colistin (93.4%). Tigecycline was active against all resistant subsets of the Enterobacterales (MIC90, 1-4 mg/L) and among Escherichia coli isolates, > 90% from each resistant subset were susceptible to tigecycline. More than 99% of all P. aeruginosa isolates, including MDR isolates and the carbapenemase-positive, MBL-positive subset, were susceptible to colistin.

METHODS: A total of 378 AMR-ESKAPEE strains were obtained based on convenience sampling over a nine-month study period (2019-2020). All strains were subjected to disk diffusion and broth microdilution assays to determine the antimicrobial susceptibility profiles. Polymerase chain reaction (PCR) and DNA sequence analyses were performed to determine the AMR genes profiles of the non-susceptible strains. Chi-square test and logistic regression analyses were used to correlate the AMR profiles and clinical data to determine the risk factors associated with HAIs.

RESULTS: High rates of multidrug resistance (MDR) were observed in A. baumannii, K. pneumoniae, E. coli, and S. aureus (69-89%). All organisms except E. coli were frequently associated with HAIs (61-94%). Non-susceptibility to the last-resort drugs vancomycin (in Enterococcus spp. and S. aureus), carbapenems (in A. baumannii, P. aeruginosa, and Enterobacteriaceae), and colistin (in Enterobacteriaceae) were observed. Both A. baumannii and K. pneumoniae harbored a wide array of extended-spectrum β-lactamase genes (blaTEM, blaSHV, blaCTX-M, blaOXA). Metallo-β-lactamase genes (blaVEB, blaVIM, blaNDM) were detected in carbapenem-resistant strains, at a higher frequency compared to other local reports. We detected two novel mutations in the quinolone-resistant determining region of the gyrA in fluoroquinolone-resistant E. coli (Leu-102-Ala; Gly-105-Val). Microbial resistance to ampicillin, methicillin, and cephalosporins was identified as important risk factors associated with HAIs in the hospital.

RESULTS: A novel approach of utilizing an in-trans protein surface display system of Lactobacillus plantarum carrying and displaying combination of Mycobacterium tuberculosis subunit epitope antigens (Ag85B, CFP-10, ESAT-6, Rv0475 and Rv2031c) fused with LysM anchor motif designated as ACERL was constructed, cloned and expressed in Esherichia coli Rossetta expression host. Subsequently the binding capability of ACERL to the cell wall of L. plantarum was examined via the immunofluorescence microscopy and whole cell ELISA where successful attachment and consistent stability of cell wall binding up to 4 days was determined. The immunization of the developed vaccine of L. plantarum surface displaying ACERL (Lp ACERL) via the oral route was studied in mice for its immunogenicity effects. Lp ACERL immunization was able to invoke significant immune responses that favor the Th1 type cytokine response of IFN-γ, IL-12 and IL-2 as indicated by the outcome from the cytokine profiling of spleen, lung, gastrointestinal tract (GIT), and the re-stimulation of the splenocytes from the immunized mice. Co-administration of an adjuvant consisting of Lactococcus lactis secreting mouse IL-12 (LcIL-12) with Lp ACERL was also investigated. It was shown that the addition of LcIL-12 was able to further generate significant Th1 type cytokines immune responses, similar or better than that of Lp ACERL alone which can be observed from the cytokine profiling of the immunized mice's spleen, lung and GIT.

RESULT: The screening of six physical conditions by Plackett-Burman Design has identified pH, inoculum size and incubation time as exerting significant effects on lipase production. These three conditions were further optimised using, Box-Behnken Design of Response Surface Methodology, which predicted an optimum medium comprising pH 6, 24 h incubation time and 2% inoculum size. T1 lipase activity of 2.0 U/mL was produced with a biomass of OD600 23.0.