Molecular Epidemiological Characterisation of ESBL- and Plasmid-Mediated AmpC-Producing Escherichia coli and Klebsiella pneumoniae at Kamuzu Central Hospital, Lilongwe, Malawi.

The global rise in infections caused by multidrug resistant (MDR) Enterobacterales poses a public health problem. We have performed a molecular epidemiological characterisation of representative plasmid-mediated AmpC (pAmpC) and ESBL-positive clinical isolates of Escherichia coli (n = 38) and Klebsiella pneumoniae (n = 17) from a tertiary hospital in Malawi collected in 2017. BlaCTX-M-15 was the most prevalent ESBL-determinant in E. coli (n = 30/38) and K. pneumoniae (n = 17/17), whereas blaCMY-2 was detected in nearly all AmpC-phenotype E. coli (n = 15/17). Whole genome sequencing revealed dominant globally disseminated E. coli sequence types (STs); ST410 (n = 16), ST131 (n = 7), and ST617 (n = 6). The ST distribution in K. pneumoniae was more diverse but included ST101 (n = 2), ST14 (n = 2), and ST340 (n = 2), all considered high-risk MDR clones. The isolates expressed an MDR profile, including resistance against commonly used antibiotics, such as fluoroquinolones, aminoglycosides, and/or trimethoprim-sulfamethoxazole, and harboured corresponding resistance determinants. Clonal analyses of the major STs of E. coli revealed closely related genetic clusters within ST410, ST131, and ST617 supporting within-hospital transmission between patients and/or via a common reservoir. The overall findings add to the limited knowledge on the molecular epidemiology of MDR E. coli and K. pneumoniae in Malawi and may help health policy makers to identify areas to target when addressing this major threat of antibiotic resistance.

Antimicrobial susceptibility profiles of clinically important bacterial pathogens at the Kamuzu Central Hospital in Lilongwe, Malawi.

Background: The aim of this prospective study was to ascertain antimicrobial resistance (AMR) in clinical bacterial pathogens from in-hospital adult patients at a tertiary hospital in Lilongwe, Malawi. Methods: Clinical specimens (blood culture, pus, urine and cerebrospinal fluid) collected during June to December 2017 were examined for bacterial growth in standard aerobic conditions. One specimen per patient was included. Antimicrobial susceptibility testing (AST) was performed using the disk diffusion method and interpreted according to EUCAST guidelines. Results: A total of 694 specimens were collected during the study period, of which 336 (48%) specimen yielded visible bacterial growth. Of the 336 specimens, a total of 411 phenotypically different isolates were recovered. Of the 411 isolates, 84 isolates (20%) were excluded and the remaining 327 (80%) were further characterised. The characterised isolates were identified as ESKAPE pathogens (n=195/327; 60%), Escherichia coli (n=92/327; 28%), Proteus mirabilis (n=33/327; 10) or Salmonella spp. (n=7/327; 2%) and were included for further analysis. The excluded isolates (n=84) comprised of coagulase-negative staphylococci (n=25), streptococci (n=33), and low-prevalence Gram-negative bacilli (n=26). E. coli (n=92; 28%) and S. aureus (n=86; 26%) were the most dominant species. A multidrug resistant (MDR) extended spectrum ß- lactamase (ESBL)-positive phenotype was detected in Klebsiella pneumoniae (n=20/29; 69%) and E. coli (n=49/92; 53%). One third of the Pseudomonas aeruginosa isolates were resistant to meropenem (MEM), but did not appear to be carbapenemase-producers. Methicillin resistant Staphylococcus aureus (MRSA) was molecularly confirmed in 10.5% of S. aureus (n=9/86). Conclusion: The high proportion of the MDR ESBL-phenotype in clinical isolates of Enterobacterales, strongly limits antimicrobial treatment options and has consequences for empirical and targeted antimicrobial treatment as well as clinical microbiology services and hospital infection control. There is need for a continuous surveillance and an antimicrobial stewardship (AMS) program to contain and prevent the spread of AMR.

Genomic epidemiology of Escherichia coli isolates from a tertiary referral center in Lilongwe, Malawi.

Antimicrobial resistance (AMR) is a global threat, including in sub-Saharan Africa. However, little is known about the genetics of resistant bacteria in the region. In Malawi, there is growing concern about increasing rates of antimicrobial resistance to most empirically used antimicrobials. The highly drug resistant Escherichia coli sequence type (ST) 131, which is associated with the extended spectrum ß-lactamase blaCTX-M-15, has been increasing in prevalence globally. Previous data from isolates collected between 2006 and 2013 in southern Malawi have revealed the presence of ST131 and the blaCTX-M-15 gene in the country. We performed whole genome sequencing (WGS) of 58 clinical E. coli isolates at Kamuzu Central Hospital, a tertiary care centre in central Malawi, collected from 2012 to 2018. We used Oxford Nanopore Technologies (ONT) sequencing, which was performed in Malawi. We show that ST131 is observed more often (14.9% increasing to 32.8%) and that the blaCTX-M-15 gene is occurring at a higher frequency (21.3% increasing to 44.8%). Phylogenetics indicates that isolates are highly related between the central and southern geographic regions and confirms that ST131 isolates are contained in a single group. All AMR genes, including blaCTX-M-15, were widely distributed across sequence types. We also identified an increased number of ST410 isolates, which in this study tend to carry a plasmid-located copy of blaCTX-M-15 gene at a higher frequency than blaCTX-M-15 occurs in ST131. This study confirms the expanding nature of ST131 and the wide distribution of the blaCTX-M-15 gene in Malawi. We also highlight the feasibility of conducting longitudinal genomic epidemiology studies of important bacteria with the sequencing done on site using a nanopore platform that requires minimal infrastructure.