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.
Resistance to disinfectants is well investigated in staphylococci and pseudomonads but nearly unexplored in bacteria of the genus Enterococcus, despite their rising significance as nosocomial pathogens. In this study, Enterococcus faecalis (n=585) from blood (n=42) and stool (n=109) of hospitalized humans, from faeces of farm animals (n=226), and from food (milk and dairy products, n=96; meat and meat products, n=112) were screened for the presence of qac-genes (qacA, qacB, qacC, smr [qacC+qacD], qacEΔ1, qacG, qacH, qacJ) via PCR. The isolates' susceptibility to a quaternary ammonium compound (didecyldimethylammoniumchloride, DDAC) and antibiotics was assessed by microdilution. Four E. faecalis strains were positive for qac-genes: qacA/B was found in one isolate from cattle and one isolate from human blood; smr (qacC+qacD) was detected in one isolate from human stool and in one isolate from cheese ("Camembert"). The sequences of the qacA/B-amplicons differed in two basepairs. DDAC had an elevated minimum inhibitory concentration (MIC) of 2.45-3.5 mg/L in one qacA/B-positive strain from human blood, whereas the other qac-gene carriers had wild-type MIC-values for DDAC (1.05 mg/L). This is the first detection of qacA/B in the genus Enterococcus.
Ampicillin and vancomycin are important antibiotics for the therapy of Enterococcus faecalis infections. The ampicillin resistance gene pbp5 is intrinsic in Enterococcus faecium. The vanC1 gene confers resistance to vancomycin and serves as a species marker for Enterococcus gallinarum. Both genes are chromosomally located. Resistance to ampicillin and vancomycin was determined in 484 E. faecalis of human and porcine origin by microdilution. Since E. faecalis are highly skilled to acquire resistance genes, all strains were investigated for the presence of pbp5 (and, in positive strains, for the penicillin-binding protein synthesis repressor gene psr) and vanC1 (and, in positive strains, for vanXYc and vanT) by using polymerase chain reaction (PCR). One porcine and one human isolate were phenotypically resistant to ampicillin; no strain was vancomycin resistant. Four E. faecalis (3/1 of porcine/human origin) carried pbp5 (MIC=1 mg/L), and four porcine strains were vanC1 positive (minimum inhibitory concentration [MIC]=1 mg/L). Real-time reverse transcriptase (RT)-PCR revealed that the genes were not expressed. The psr gene was absent in the four pbp5-positive strains; the vanXYc gene was absent in the four vanC1-positive strains. However, vanT of the vanC gene cluster was detected in two vanC1-positive strains. To our knowledge, this is the first report on the presence of pbp5, identical with the "E. faecium pbp5 gene," and of vanC1/vanT in E. faecalis. Even if resistance is not expressed in these strains, this study shows that E. faecalis have a strong ability to acquire resistance genes-and potentially to spread them to other bacteria. Therefore, close monitoring of this species should be continued.
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