Insusceptibility to disinfectants in bacteria from animals, food and humans-is there a link to antimicrobial resistance?

Abstract

Enterococcus faecalis (n = 834) and Enterococcus faecium (n = 135) from blood and feces of hospitalized humans, from feces of outpatients and livestock and from food were screened for their susceptibility to a quaternary ammonium compound (didecyldimethylammoniumchloride, DDAC) and to 28 antibiotics by micro-/macrodilution. The maximum DDAC-MIC in our field study was 3.5 mg/l, but after adaptation in the laboratory, MIC values of 21.9 mg/l were observed. Strains for which DDAC had MICs > 1.4 mg/l ("non-wildtype," in total: 46 of 969 isolates/4.7%) were most often found in milk and dairy products (14.6%), while their prevalence in livestock was generally low (0-4%). Of human isolates, 2.9-6.8% had a "non-wildtype" phenotype. An association between reduced susceptibility to DDAC, high-level-aminoglycoside resistance and aminopenicillin resistance was seen in E. faecium (p < 0.05). No indications for a common source of non-wildtype strains were found by RAPD-PCR; however, several non-wildtype E. faecalis shared the same variant of the emeA-gene. In addition, bacteria (n = 42) of different genera were isolated from formic acid based boot bath disinfectant (20 ml of 55% formic acid/l). The MICs of this disinfectant exceeded the wildtype MICs up to 20-fold (staphylococci), but were still one to three orders of magnitude below the used concentration of the disinfectant (i. e., 1.1% formic acid). In conclusion, the bacterial susceptibility to disinfectants still seems to be high. Thus, the proper use of disinfectants in livestock surroundings along with a good hygiene praxis should still be highly encouraged. Hints to a link between antibiotic resistance and reduced susceptibility for disinfectants-as seen for E. faecium-should be substantiated in further studies and might be an additional reason to confine the use of antibiotics.

Keywords: QAC; aminoglycoside; antimicrobial resistance; antiseptic; biocide; disinfectant; enterococci; formic acid.

Figure 1

MIC-values (log2) of didecyldimethylammoniumchloride (DDAC) in E. faecalis with initial MICs > 1.4 mg/l, compared with control DSM 2570 during 70 days of stepwise adaptation.

Figure 2

Amplicon-typing (RAPD-PCR) of E. faecalis (MIC of DDAC > 1.4 mg/l) from different sources.

Figure 3

Amplicon-typing (RAPD-PCR, two different conditions) of E. faecalis (MIC of DDAC > 1.4 mg/l) from different sources.

Figure 4

Conventional melting curve analysis (Lightcycler I.I, SYBR-Green) of emeA-amplicons resulting in two different emeA types: type 1 (higher Tm) and type 2 (lower Tm).