A modification of the Winter and Sandholzer media and technique for enterococci detection.
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Enterococcal isolates are increasingly responsible for nosocomial infections in the last decade and they are recently cited as being the second most common pathogen isolated from hospitalized patients. Enterococcal infections can be localized at different sites: endocarditis, CNS, intrabdominal, pelvic. The epidemiological trend may be a consequence of predisposing risk factors due to hospitalization such as instrumentation, immunosuppression, long-term hospitalization. Nevertheless the most alarming aspect of the problem is the increasing detection of antibiotic resistance Enterococcal strains, especially the high level resistance (HLR) to aminoglycosides, penicillin and glycopeptides must not be underestimated. Optimal antibiotic regimes for the treatment of multiple resistant strains in severe enterococcal infections are not been defined yet and antibiotic association such as aminoglycosides + glycopeptides or glycopeptides + fluoroquinolones could be suggested.
According to new reports the AAC (6')-APH (2")Ia gene is no longer the only gene encoding resistance to gentamycin in Gram-positive cocci and therefore the current method for predicting synergism aminoglycosides with bacterial cell wall active agents in this bacteria may need revision. To further our knowledge of aminoglycoside resistance mechanism in Gram-positive cocci in Gdańsk region we tested presence of AAC (6')-APH (2")Ia gene among 22 enterococcal (E. faecalis) and 41 staphylococcal (S. haemolyticus, S. aureus, S. epidermidis) gentamycin-resistant isolates. Presence of AAC (6')-APH (2")Ia gene varied from 50% (n = 6) in gentamycin-resistant S. epidermidis, 80% (n = 10) in gentamycin resistant S. haemolyticus 88% in methicillin-resistant Staphylococcus aureus (MRSA) (n = 25). In Enterococcus faecalis this gene was noticed only in 59% (n = 22) of gentamycin-resistant isolates. These results suggest that spread of resistance gene among different species is limited and AAC (6')-APH (2")Ia mediated gentamycin-resistance mechanism is more common among MRSA and Staphylococcus haemolyticus.
The present study proposed use of modified multilocus sequence typing (MLST) approach, for subspecies differentiation of Enterococcus faecalis isolates. We disgesed with SspI endonuclease product of amplification of three E. faecalis antigen-encoding genes (ace, encoding a collagen and laminin adhesin; efaA, encoding an endocarditis antigen; and salA, encoding a cell wall associated antigen) and one housekeeping gene (pyrC) of 32 E. faecalis isolates MLRA analysis of polymorphic regions of these four genes identified 23 distinct types 21.4 to 100% identity among the 32 isolates. Further studies are needed to evaluate the value of proposed method.
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A study on the antibiotic resistance of enterococcal isolates (n = 280) was carried out in a teaching hospital in Naples. Strains were isolated from different sources, identified by conventional tests and their antibiotic susceptibility was tested by E-test method. Thirty-two enterococcal isolates (11.5%), phenotypically identified as E. faecium (n = 26), E. gallinarum (n = 3), E. faecalis (n = 2) and E. hirae (n = 1), showed resistance to glycopeptides. The vanA gene was found in all 32 VRE. Molecular typing was performed by RAPD analysis which showed two majors patterns.
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Combinations of one of the new fluoroquinolones (ciprofloxacin, temafloxacin or tosufloxacin) with a betalactam (amoxicillin, piperacillin, or imipenem) were tested in vitro using the checkboard method against 28 strains of Streptococcus faecalis recovered in 1990 from a variety of specimens. No instance of antagonism (FIC greater than 2) was recorded. Effects of the two agents were usually additive (0.5 less than FIC less than or equal to 1). An indifferent effect (1 less than FIC less than or equal to 2) was seen in 32.1% of cases (9 strains) with the amoxicillin-tosufloxacin combination. Synergy (FIC less than or equal to 0.5) was uncommon with combinations including imipenem: effects were synergistic for only three strains with imipenem and either ciprofloxacin or tosufloxacin and for six strains with the imipenem-temafloxacin combination. Synergy was more common with combinations including ciprofloxacin and either piperacillin (10 strains, 35.7%) or amoxicillin (13 strains, 46.4%).
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