BackgroundVancomycin-resistant isolates of E. faecalis and E. faecium are of special concern and patients at risk of acquiring a VRE colonization/infection include also intensively-cared neonates. We describe here an ongoing high prevalence of VanB type E. faecium in a neonatal ICU hardly to identify by routine diagnostics.MethodsDuring a 10 months’ key period 71 E. faecium isolates including 67 vanB-type isolates from 61 patients were collected non-selectively. Vancomycin resistance was determined by different MIC methods (broth microdilution, Vitek® 2) including two Etest® protocols (McFarland 0.5/2.0. on Mueller-Hinton/Brain Heart Infusion agars). Performance of three chromogenic VRE agars to identify the vanB type outbreak VRE was evaluated (BrillianceTM VRE agar, chromIDTM VRE agar, CHROMagarTM VRE). Isolates were genotyped by SmaI- and CeuI-macrorestriction analysis in PFGE, plasmid profiling, vanB Southern hybridisations as well as MLST typing.ResultsMajority of vanB isolates (n = 56, 79%) belonged to a single ST192 outbreak strain type showing an identical PFGE pattern and analyzed representative isolates revealed a chromosomal localization of a vanB2-Tn5382 cluster type. Vancomycin MICs in cation-adjusted MH broth revealed a susceptible value of ≤4 mg/L for 31 (55%) of the 56 outbreak VRE isolates. Etest® vancomycin on MH and BHI agars revealed only two vanB VRE isolates with a susceptible result; in general Etest® MIC results were about 1 to 2 doubling dilutions higher than MICs assessed in broth and values after the 48 h readout were 0.5 to 1 doubling dilutions higher for vanB VRE. Of all vanB type VRE only three, three and two isolates did not grow on BrillianceTM VRE agar, chromIDTM VRE agar and CHROMagarTM VRE, respectively. Permanent cross contamination via the patients’ surrounding appeared as a possible risk factor for permanent VRE colonization/infection.ConclusionsLow level expression of vanB resistance may complicate a proper routine diagnostics of vanB VRE and mask an ongoing high VRE prevalence. A high inoculum and growth on rich solid media showed the highest sensitivity in identifying vanB type resistance.

Many surveillance methods for nosocomial infections (NIs) have been put forward in the literature, and all have their advantages and disadvantages. Different surveillance methods are useful, depending on whether the objective of surveillance is only to increase sensitivity to infection control problems and to identify areas with possible infection control problems; to confirm a possible infection control problem through comparison with other units or departments; or to use surveillance data for identifying the sources of infections. Furthermore, time effectiveness is a major point in selecting the most appropriate method, particularly the method for case identification. In units or departments with a high level of NI, even highly time-consuming surveillance methods may be ultimately time-effective; in units or departments with a lower level of NI, the time-effectiveness depends on the time necessary for case identification. Close liaison with staff in the units is a sine qua non for the success of all surveillance activities.

A 20-ml blood sample was collected from adult patients with suspected bloodstream infections and distributed equally into the four volume-controlled bottles of a blood culture set consisting of aerobic and anaerobic BACTEC Plus/F bottles and aerobic and anaerobic BacT/Alert FAN bottles. All bottles were incubated in their respective instruments for a standard 5-day protocol or until the instruments signalled positivity. Samples in all bottles with negative results by these instruments were terminally subcultured. A total of 8,390 blood culture sets were obtained during the study period, of which 4,402 (52.5%) met the study criteria. Of these, 946 (21.5%) were positive either by instrument signal or by additional terminal subculture of all negative bottles and yielded growth of microorganisms. Five hundred eighty-nine (13.4%) blood culture sets were considered to have recovered 663 clinically significant organisms. When both the BACTEC and the BacT/Alert systems were used, 465 positive sets were detected; BACTEC alone detected 52 positive sets and BacT/Alert alone detected 72 (P = 0.09). No differences were found between the two systems in microbial recovery rate from blood cultures obtained from patients on antibiotic therapy. Significantly more members of the family Enterobacteriaceae (P < 0.01) were detected from patients without antimicrobial therapy by BacT/Alert than by BACTEC. The false-negative rates were 0.20% for BACTEC and 0.32% for BacT/Alert. A significantly higher false-positive rate was found for BACTEC (P < 0.0001). Both systems were comparable for the time to detection of microorganisms. However, gram-positive bacteria were detected faster by BACTEC andEnterobacteriaceae were detected faster on average by BacT/Alert. We concluded that both systems are comparable in their abilities to recover aerobic and anaerobic organisms from blood cultures and a terminal subculture might not be necessary for either of the two systems. The increased positivity rate when using an anaerobic bottle in a two-bottle blood culture set is due to the additional blood volume rather than to the use of an anaerobic medium.

In a study comparing the Bactec 9240 (Plus Aerobic/F and Anaerobic/F bottles, containing resins; Becton Dickinson, USA) and the Vital (standard aerobic and anaerobic bottles, with no additives; bioMérieux, France) blood culture systems, 6456 sets of four bottles of 9660 blood cultures submitted were evaluated. There were 531 clinically significant isolates from 795 positive blood cultures. Of the 531 positive blood cultures, 355 were positive in both systems, 141 with the Bactec 9240 alone, and 30 with the Vital alone (p < 0.001); five were not detected by either system. The average time to detection of positive cultures for the matched sets was 10.65 h and 18.41 h by the Bactec 9240 system and the Vital system, respectively. The false-positive rate per bottle was 0.65% in the Bactec 9240 and 0.71% in the Vital. The rate of false-negative pairs (i.e., major errors) was very low (0.12% for the Bactec 9240, 0.19% for the Vital) and not significantly different between the two systems. The striking differences in recovery of microorganisms may be due to the presence of resins in the Bactec medium. However, the observed superiority of the Bactec 9240, even for patients not receiving antibiotics, suggests that resins adsorb other inhibitors present in patients' blood.

Although there is a 20% yeast colonization in the gastrointestinal tract of the population, fungal infections appear only rarely in secondary peritonitis. The risk of severe mycosis increases after a major operation and when a patient is taking broad-spectrum antibiotics, is on total parenteral nutrition, is catheterized, and/or is immune-suppressed. In the past years the incidence of nosocomial fungal infections (usually Candida spp.) has risen significantly. Five percent of CAPD-related peritonitis is caused by fungi. In enteral anastomosis breakdown, invasive mycosis occurs more often, with an accompanying lethality of up to 80%. In severe pancreatitis, up to 5% of peripancreatic necrosis is infected with fungi. The clinical course of severe mycosis, like the septic syndrome, is associated with fungemia in up to 50% of cases. As most of the facultative pathogenic fungi are part of the physiological flora, it is difficult to interpret mycological cultures. In order to diagnose invasive fungal infections, histopathological techniques and serologic tests for antigens and antibodies are available. Three antifungal agents (amphotericin B, flucytosine, fluconazole) are available for intravenous administration. Amphotericin B is given at doses of up to 1 mg/kg per day, in liposomal galenism up to 3 mg/kg per day. Combining amphotericin B with flucytosine (150-200 mg/kg per day) a synergistic effect is reached. Fluconazole at a dosage of 200-800 mg per day represents an alternative with similar antifungal activity and lower side effects.