Enterobacter cloacae harbouring blaNDM-1, blaKPC , and blaOXA-48-like  carbapenem-resistant genes isolated from different environmental sources in South Africa

Abstract Background The emergence of carbapenem-resistant and colistin-resistant ECC pose a huge challenge to infection control. The purpose of this study was to clarify the mechanism of the carbapenems and colistin co-resistance in Enterobacter cloacae Complex (ECC) strains. Results This study showed that the mechanisms of carbapenem resistance in this study are: 1. Generating carbapenemase (7 of 19); 2. The production of AmpC or ESBLs combined with decreased expression of out membrane protein (12 of 19). hsp60 sequence analysis suggested 10 of 19 the strains belong to colistin hetero-resistant clusters and the mechanism of colistin resistance is increasing expression of acrA in the efflux pump AcrAB-TolC alone (18 of 19) or accompanied by a decrease of affinity between colistin and outer membrane caused by the modification of lipid A (14 of 19). Moreover, an ECC strain co-harboring plasmid-mediated mcr-4.3 and blaNDM-1 has been found. Conclusions This study suggested that there is no overlap between the resistance mechanism of co-resistant ECC strains to carbapenem and colistin. However, the emergence of strain co-harboring plasmid-mediated resistance genes indicated that ECC is a potential carrier for the horizontal spread of carbapenems and colistin resistance.

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Enterobacteriaceae isolates carrying the New Delhi metallo-β-lactamase gene in Yemen

Journal of Medical Microbiology ◽

10.1099/jmm.0.073767-0 ◽

2014 ◽

Vol 63 (10) ◽

pp. 1316-1323 ◽

Cited By ~ 24

Author(s):

Alima Gharout-Sait ◽

Samer-Ahmed Alsharapy ◽

Lucien Brasme ◽

Abdelaziz Touati ◽

Rachida Kermas ◽

...

Keyword(s):

Klebsiella Pneumoniae ◽

Susceptibility Testing ◽

Enterobacter Cloacae ◽

New Delhi ◽

Carbapenem Resistant ◽

Genes Encoding ◽

Phenotypic Tests ◽

Lactamase Gene ◽

Sequence Types

Ten carbapenem-resistant Enterobacteriaceae (eight Klebsiella pneumoniae isolates and two Enterobacter cloacae) isolates from Yemen were investigated using in vitro antimicrobial susceptibility testing, phenotypic carbapenemase detection, multilocus sequence typing (MLST) and replicon typing. Carbapenemase, extended-spectrum β-lactamase (ESBL) and plasmid-mediated quinolone resistance determinant genes were identified using PCR and sequencing. All of the 10 carbapenem-resistant Enterobacteriaceae were resistant to β-lactams, tobramycin, ciprofloxacin and cotrimoxazole. Imipenem, doripenem and meropenem MICs ranged from 2 to >32 mg l−1 and ertapenem MICs ranged from 6 to >32 mg l−1. All of the K. pneumoniae isolates showed ESBL activity in phenotypic tests. Genes encoding bla NDM were detected in all strains. All K. pneumoniae strains produced CTX-M-15 ESBL and SHV β-lactamases. TEM-1 β-lactamase was detected in seven isolates. Nine isolates were qnr positive including QnrB1, QnrA1 and QnrS1, and six isolates produced AAC-6′-Ib-cr. MLST identified five different sequence types (STs): ST1399, ST147, ST29, ST405 and ST340. Replicon typing showed the presence of IncFII1K plasmids in four transformants. To the best of our knowledge, this is the first report of NDM-1-producing Enterobacteriaceae isolates in Yemen.

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Genomic Investigation of a Putative Endoscope-Associated Carbapenem-Resistant Enterobacter cloacae Outbreak Reveals a Wide Diversity of Circulating Strains and Resistance Mutations

Clinical Infectious Diseases ◽

10.1093/cid/cix934 ◽

2017 ◽

Vol 66 (3) ◽

pp. 460-463 ◽

Cited By ~ 4

Author(s):

Shawn E Hawken ◽

Laraine L Washer ◽

Christopher L Williams ◽

Duane W Newton ◽

Evan S Snitkin

Keyword(s):

Enterobacter Cloacae ◽

Resistance Mutations ◽

Carbapenem Resistant

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Comparative Pathogenomics of Aeromonas veronii from Pigs in South Africa: Dominance of the Novel ST657 Clone

Microorganisms ◽

10.3390/microorganisms8122008 ◽

2020 ◽

Vol 8 (12) ◽

pp. 2008

Author(s):

Yogandree Ramsamy ◽

Koleka P. Mlisana ◽

Daniel G. Amoako ◽

Akebe Luther King Abia ◽

Mushal Allam ◽

...

Keyword(s):

South Africa ◽

Orthologous Gene ◽

Gene Clusters ◽

Illumina Miseq ◽

Aeromonas Veronii ◽

Carbapenem Resistant ◽

Kwazulu Natal ◽

Miseq Platform ◽

Differential Ability ◽

Potential Pathogenicity

The pathogenomics of carbapenem-resistant Aeromonas veronii (A. veronii) isolates recovered from pigs in KwaZulu-Natal, South Africa, was explored by whole genome sequencing on the Illumina MiSeq platform. Genomic functional annotation revealed a vast array of similar central networks (metabolic, cellular, and biochemical). The pan-genome analysis showed that the isolates formed a total of 4349 orthologous gene clusters, 4296 of which were shared; no unique clusters were observed. All the isolates had similar resistance phenotypes, which corroborated their chromosomally mediated resistome (blaCPHA3 and blaOXA-12) and belonged to a novel sequence type, ST657 (a satellite clone). Isolates in the same sub-clades clustered according to their clonal lineages and host. Mobilome analysis revealed the presence of chromosome-borne insertion sequence families. The estimated pathogenicity score (Pscore ≈ 0.60) indicated their potential pathogenicity in humans. Furthermore, these isolates carried several virulence factors (adherence factors, toxins, and immune evasion), in different permutations and combinations, indicating a differential ability to establish infection. Phylogenomic and metadata analyses revealed a predilection for water environments and aquatic animals, with more recent reports in humans and food animals across geographies, making A. veronii a potential One Health indicator bacterium.

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Molecular Characterization of Carbapenem-Resistant Enterobacter cloacae in 11 Chinese Cities

Frontiers in Microbiology ◽

10.3389/fmicb.2018.01597 ◽

2018 ◽

Vol 9 ◽

Cited By ~ 9

Author(s):

Chunmei Jin ◽

Jiangang Zhang ◽

Qi Wang ◽

Hongbin Chen ◽

Xiaojuan Wang ◽

...

Keyword(s):

Molecular Characterization ◽

Enterobacter Cloacae ◽

Chinese Cities ◽

Carbapenem Resistant

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Detection of plasmid-mediated colistin-resistant and carbapenem-resistant genes by multiplex PCR

MethodsX ◽

10.1016/j.mex.2018.05.016 ◽

2018 ◽

Vol 5 ◽

pp. 532-536 ◽

Cited By ~ 4

Author(s):

Rujirat Hatrongjit ◽

Anusak Kerdsin ◽

Yukihiro Akeda ◽

Shigeyuki Hamada

Keyword(s):

Multiplex Pcr ◽

Carbapenem Resistant ◽

Resistant Genes

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In Vitro Antibacterial Activity and Mechanism of Vanillic Acid against Carbapenem-Resistant Enterobacter cloacae

Antibiotics ◽

10.3390/antibiotics8040220 ◽

2019 ◽

Vol 8 (4) ◽

pp. 220 ◽

Cited By ~ 5

Author(s):

Weidong Qian ◽

Yuting Fu ◽

Miao Liu ◽

Ting Wang ◽

Jianing Zhang ◽

...

Keyword(s):

Antibacterial Activity ◽

Membrane Potential ◽

Cell Membrane ◽

Crystal Violet ◽

Vanillic Acid ◽

Membrane Integrity ◽

Enterobacter Cloacae ◽

Carbapenem Resistant ◽

Atp Concentration

Vanillic acid (VA) is a flavoring agent found in edible plants and fruits. Few recent studies exhibited robust antibacterial activity of VA against several pathogen microorganisms. However, little was reported about the effect of VA on carbapenem-resistant Enterobacter cloacae (CREC). The purpose of the current study was to assess in vitro antimicrobial and antibiofilm activities of VA against CREC. Here, minimum inhibitory concentrations (MIC) of VA against CREC was determined via gradient diffusion method. Furthermore, the antibacterial mode of VA against CREC was elucidated by measuring changes in intracellular adenosine triphosphate (ATP) concentration, intracellular pH (pHin), cell membrane potential and membrane integrity. In addition, antibiofilm formation of VA was measured by crystal violet assay and visualized with field emission scanning electron microscopy (FESEM) and confocal laser scanning microscopy (CLSM). The results showed that MIC of VA against E. cloacae was 600 μg/mL. VA was capable of inhibiting the growth of CREC and destroying the cell membrane integrity of CREC, as confirmed by the decrease of intracellular ATP concentration, pHin and membrane potential as well as distinctive variation in cellular morphology. Moreover, crystal violet staining, FESEM and CLSM results indicated that VA displayed robust inhibitory effects on biofilm formation of CREC and inactivated biofilm-related CREC cells. These findings revealed that VA exhibits potent antibacterial activity against CREC, and thus has potential to be exploited as a natural preservative to control the CREC associated infections.

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Pathogenomic Analysis of a Novel Extensively Drug-Resistant Citrobacter freundii Isolate Carrying a blaNDM-1 Carbapenemase in South Africa

Pathogens ◽

10.3390/pathogens9020089 ◽

2020 ◽

Vol 9 (2) ◽

pp. 89 ◽

Cited By ~ 2

Author(s):

Yogandree Ramsamy ◽

Koleka P. Mlisana ◽

Daniel G. Amoako ◽

Mushal Allam ◽

Arshad Ismail ◽

...

Keyword(s):

South Africa ◽

Acquired Resistance ◽

Carbapenem Resistance ◽

Illumina Miseq ◽

Tertiary Hospital ◽

Phylogenomic Analysis ◽

Citrobacter Freundii ◽

Class 1 Integron ◽

Pathogenic Potential ◽

Carbapenem Resistant

Pathogenomic analysis was performed on a novel carbapenem-resistant Citrobacter freundii isolate (H2730R) from a rectal swab of an adult male patient admitted to a tertiary hospital, Durban, South Africa. H2730R was identified using selective media and API 20e kit. Confirmatory identification and antibiotic susceptibility testing were performed using the VITEK II. H2730R was whole-genome sequenced on the Illumina MiSeq platform. H2730R was resistant to all tested antibiotics except tigecycline and was defined as ST498 by the C. freundii multilocus sequence typing (MLST) database. The estimated pathogenic potential predicted a higher probability (Pscore ≈ 0.875), supporting H2730R as a human pathogen. H2730R harbored 25 putative acquired resistance genes, 4 plasmid replicons, 4 intact prophages, a class 1 integron (IntI1), 2 predominant insertion sequences (IS3 and IS5), numerous efflux genes, and virulome. BLASTn analysis of the blaNDM-1 encoding contig (00022) and its flanking sequences revealed the blaNDM-1 was located on a plasmid similar to the multireplicon p18-43_01 plasmid reported for the spread of carbapenem resistance in South Africa. Phylogenomic analysis showed clustering of H2730R with CF003/CF004 strains in the same clade, suggesting a possible association between C. freundii strains/clones. Acquiring the p18-43_01 plasmid containing blaNDM-1, the diversity, and complex resistome, virulome, and mobilome of this pathogen makes its incidence very worrying regarding mobilized resistance. This study presents the background genomic information for future surveillance and tracking of the spread of carbapenem-resistant Enterobacteriaceae in South Africa.

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