A comparative study for detection of extended spectrum β-lactamase (ESBL) production by Enteroaggregative Escherichia coli (EAEC) strains using double disc, nitrocefin and PCR assays

Vergis, Jess; Pathak, Richa; Kumar, Manesh; Ramanjeneya, Sunitha; Malik, S.V.S.; Barbuddhe, Sukhadeo B.; Rawool, Deepak B.

doi:10.1016/j.mimet.2018.06.004

Cited by 4 publications

(3 citation statements)

References 37 publications

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“…This may be due to a low betalactamase activity and the MIC value is less than necessary to generate resistance. It has also been described that the nitrocefin test can generate false positives or negatives because many times the molecular reactions are incomplete (Pitkälä et al, 2007), although an effectiveness greater than 90 % has been described (Vergis et al, 2018). It would be interesting to sequence the genome of this strain to identify the beta-lactamase involved.…”

Section: Discussionmentioning

confidence: 99%

Antibiotic Resistance of Enterococcus faecalis from Persistent Endodontic Infections

Seguel¹,

Quezada-Aguiluz

González-Rocha

et al. 2020

Int. J. Odontostomat.

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Enterococci are important nosocomial pathogens due to their intrinsic multiresistance and the acquisition of new antibiotic resistance genes (ARG). Enterococcus faecalis has been shown to be one of the main pathogens in persistent endodontic infections, therefore, the main objective of this study was to evaluate the phenotype and resistance genotype of strains of E. faecalis isolated from teeth with persistent endodontic lesions, to the most commonly prescribed antibiotics in dentistry. Thirteen strains of E. faecalis of different pulsotype were analyzed to evaluate the susceptibility to antibiotics, amoxicillin, amoxicillin/clavulanic acid, tetracycline, erythromycin and metronidazole, using the Epsilometer test (E-test) and the presence of beta-lactamases with nitrocefin test. Finally, the detection of ARG was performed with a molecular polymerase chain reaction (PCR) technique and confirmed by the sequencing of the amplification products. Fisher's exact test was used, using 95 % confidence. Regarding the phenotype of resistance, the evaluated strains, independent of the pulsotype, were totally resistant to the action of metronidazole. Antibiotics with higher minimum inhibitory concentration (MIC) after metronidazole include tetracycline and erythromycin. In contrast, lower MIC are applied to the combination of amoxicillin with clavulanic acid. The nitrocefin test was positive only in one strain. Genotypically, two genetically distant strains isolated from a single patient, presented a genotype of resistance to erythromycin, determined by the presence of the ermB gene. No statistically significant relationship was found between phenotypic resistance and the presence of ARG in relation to erythromycin (p> 0.05). It was concluded that isolates of E. faecalis from persistent endodontic infections showed phenotypes of resistance to several antimicrobial agents, all of which were susceptible to amoxicillin/clavulanic acid. Periodic evaluation of susceptibility to antibiotics is suggested as an important practice for the surveillance of antibiotic resistance in oral strains.

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Section: Discussionmentioning

confidence: 99%

Antibiotic Resistance of Enterococcus faecalis from Persistent Endodontic Infections

Seguel¹,

Quezada-Aguiluz

González-Rocha

et al. 2020

Int. J. Odontostomat.

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“…All the EAEC strains used in this study were designated "typical," as confirmed by the classical "stacked-brick" pattern in HEp-2 adherence assay and possessed plasmid-borne transcriptional activator aggR regulon. Moreover, MDR-EAEC strains employed in this study were ESBL producers as evidenced by nitrocefin, double-disc testing as well as PCR assays (Vergis et al, 2018), highlighting an emerging trend in the drug resistance pattern against EAEC, especially toward drugs of Relative expression of the fis gene in EAEC strains was quantified by real-time PCR and normalized to the housekeeping gene, 16S rRNA, and fold changes over the non-biofilm control group were calculated by the 2 ÀΔΔCT method empirical therapy, that is, fluoroquinolones and beta-lactams (Kong et al, 2015).…”

Section: Discussionmentioning

confidence: 95%

“…The revived test strains were re‐validated using PCR employing chromosome‐ as well as plasmid‐encoded genes (Vijay et al, 2015), HEp‐2 cell adherence assay (Cravioto et al, 1979), antibiotic susceptibility testing using disc diffusion and minimum inhibitory concentration estimation (CLSI 2019). Besides, the strains were subjected to its extended‐spectrum beta‐lactamase (ESBL)‐ production using nitrocefin, double‐disc diffusion, and PCR ( blaCTXM , blaTEM , blaAmpC , and blaSHV ) assays (Vergis et al, 2018).…”

Section: Methodsmentioning

confidence: 99%

Comparative efficiency of carbohydrates on the biofilm‐forming ability of enteroaggregative Escherichia coli

Pathak

Vergis

Chouhan

et al. 2022

Journal of Food Safety

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The biofilm‐forming ability of microbes is often correlated with drug resistance; however, this association among enteroaggregative Escherichia coli (EAEC) strains remains naïve. Moreover, the investigations exploring uptake and assimilation of sugars either qualitatively or quantitatively by the biofilm‐forming EAEC strains have not yet been documented. Hence, this study investigated the role of various categories of sugars (monosaccharides, disaccharides, and polysaccharide) on the biofilm‐forming ability of three each of multi‐drug‐resistant (MDR) as well as non‐MDR typical EAEC strains. The EAEC strains exhibited black colonies on congo red agar (CRA) with monosaccharides and disaccharides, whereas all the strains exhibited red colonies on CRA with the polysaccharide. Although significant (p < 0.001) biofilm‐forming ability was observed by all the EAEC strains in sugar‐dependent microtiter plate assay, color change from pink to orange and yellow was noticed in presence of monosaccharides and disaccharide, while absent in polysaccharides. Besides, significantly higher bacterial counts were observed in presence of monosaccharides and disaccharides (p < 0.01); however, such significant bacterial counts were not noticed with the polysaccharide. Further, the real‐time PCR data exhibited comparatively higher relative expression of biofilm‐associated fis gene among EAEC strains grown in presence of monosaccharides, followed by disaccharide and polysaccharide, respectively that concurred with the flow cytometry data. No significant (p > 0.05) difference was observed in the biofilm‐forming ability between MDR‐ and non‐MDR EAEC strains. This study illustrated the role of carbohydrates on the biofilm‐forming ability of EAEC and warranted utmost attention concerning the usage of carbohydrates in food processing industries, for its incorporation as functional food additives.

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A dual-channel electrochemical biosensor enables concurrent detection of pathogens and antibiotic resistance

Gunasekaran,

Rostovsky,

Taussig

et al. 2024

Biosensors and Bioelectronics

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A comparative study for detection of extended spectrum β-lactamase (ESBL) production by Enteroaggregative Escherichia coli (EAEC) strains using double disc, nitrocefin and PCR assays

Cited by 4 publications

References 37 publications

Antibiotic Resistance of Enterococcus faecalis from Persistent Endodontic Infections

Antibiotic Resistance of Enterococcus faecalis from Persistent Endodontic Infections

Comparative efficiency of carbohydrates on the biofilm‐forming ability of enteroaggregative Escherichia coli

A dual-channel electrochemical biosensor enables concurrent detection of pathogens and antibiotic resistance

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