M. T. Blanco

Enzymatic activities of Candida tropicalis isolated from hospitalized patients

Secretion of hydrolytic enzymes is considered a virulence factor in Candida spp. Extracellular enzymatic activities in 29 clinical isolates of Candida tropicalis were analyzed by plate assays. C. tropicalis, similar to Candida albicans, showed elevated hemolytic and esterase activities. However, unlike C. albicans, low aspartyl protease and very low phospholipase activities were detected in C. tropicalis isolates.

Moxifloxacin and Biofilm Production by Coagulase-Negative Staphylococci

The in vitro activity of moxifloxacin against 41 strains of coagulase-negative staphylococci was determined. A relationship between the activity of moxifloxacin and biofilm formation was detected. Biofilm-producing strains were more resistant to moxifloxacin than biofilm-negative strains. Our global results obtained with six strains of Staphylococcus epidermidis showed that subinhibitory concentrations of moxifloxacin did not significantly modify biofilm formation. On the other hand, moxifloxacin concentrations of 2, 10, 50 and 100 × MIC produced a log decrease in viable count (included in a biofilm) of 0.20, 0.37, 1.10 and 1.69, respectively.

Exposure to Therapeutic Concentrations of Ritonavir, but Not Saquinavir, Reduces Secreted Aspartyl Proteinase of Candida parapsilosis

The effect of ritonavir and saquinavir, HIV proteinase inhibitors, on the secreted aspartyl proteinase (Sap) activity of Candida parapsilosis was studied. In a proteinase-inducing medium (yeast carbon base-bovine serum albumin), Sap activity in all clinical isolates of C. parapsilosis (n = 20) was observed at 37°C but not at 22°C. The presence of ritonavir at a concentration of 8 µg/ml produced an inhibition close to 50% albumin consumption and also delayed yeast growth; however, saquinavir did not have any effect on growth or on Sap activity. In Sabouraud broth, which does not induce Sap production, no effect was shown on yeast growth by either of the two HIV proteinase inhibitors studied.

Aspartyl proteinase, phospholipase, hemolytic activities and biofilm production of Candida albicans isolated from bronchial aspirates of ICU patients.

A correlation between mucosal colonization by Candida albicans and the subsequent development of invasive respiratory infection has been previously described. The aim of this study was to evaluate different enzymatic activities in vitro and to determine the capacity to form biofilms by 17 C. albicans isolates from bronchial aspirates of mechanically ventilated patients hospitalized in intensive care units. All the C. albicans clinical isolates tested were biofilm producers and displayed detectable levels of proteinase and hemolytic activities, although phospholipase activity was not detected in one strain. The correlation noted among the virulence factors studied suggests that the presence of more than one concurrent factor could facilitate the spread of infection.

La hidrofobicidad de la superficie celular como indicador de otros factores de virulencia en "Candida albicans"

OBJECTIVE To evaluate virulence factors involved in the adhesion process, such as cell surface hydrophobicity (CSH), adherence to plastic capacity, adherence capacity to buccal epithelial cells (BEC), and biofilm formation, in 17 strains of C. albicans isolated from bronchial aspirates of critically ill patients. METHOD The CSH of the strains of C. albicans was determined using the MATH method, a microbial adhesion to hydrocarbons test. The study of adherence to plastic was performed in microtitre plates in accordance with Christensens technique. Biofilm formation was studied in polystyrene microtitre plates, according to the method of Ramage. Adherence to BEC was evaluated by quantifying the percentage of adhered yeasts to cells. RESULTS All the strains studied showed factors directly involved in adhesion, with variability in the degree of expression among them. Medium-high levels of CSH were found in 52.9% of the strains. The percentage of strains with high values in adherence to plastic was 35.3%. The most hydrophobic strains were the most adherent to plastic, with a correlation coefficient of 0.76. Of the 12 biofilm-producing strains, 6 were high producers. These strains had also high levels of CSH and adherence to plastic, with significant results. All the strains studied adhered to BEC, with results ranging widely from 45 to 157 yeasts/100 BEC, with no significant correlation with the rest of the parameters studied, although CSH was seen to be an indispensable prior requisite for adherence to cells. CONCLUSION CSH is a variable characteristic in C. albicans and is directly related to adherence to plastic and biofilm formation. Ease in evaluating CSH permits its quantification, and could be used as an indicator of the presence of other determinants of pathogenicity.

Candida orthopsilosis fungemias in a Spanish tertiary care hospital: Incidence, epidemiology and antifungal susceptibility

BACKGROUND Few studies exist on prevalence of fungemia by Candida orthopsilosis, with variable results. AIMS To study the incidence, epidemiology and antifungal susceptibility of C. orthopsilosis strains isolated from fungemias over two years at a tertiary hospital. METHODS Candidemia episodes between June 2007 and June 2009 in a university hospital (Puerta del Mar, Cádiz, Spain) were studied. The strains initially identified as Candida parapsilosis were genotypically screened for C. parapsilosis sensu stricto, C. orthopsilosis and Candida metapsilosis, and their antifungal susceptibility was evaluated. RESULTS In this period 52 cases of candidemia were documented. Of the 19 strains originally identified as C. parapsilosis, 13 were confirmed as C. parapsilosis sensu stricto and 6 as C. orthopsilosis. Of the 52 isolates, the most frequent species were Candida albicans (30.8%), C. parapsilosis sensu stricto (25%), C. orthopsilosis, Candida tropicalis and Candida glabrata in equal numbers (11.5%). C. orthopsilosis isolates were susceptible to amphotericin B, caspofungin, voriconazole and fluconazole, with no significant differences in MIC values with C. parapsilosis sensu stricto. The source of isolates of C. orthopsilosis were neonates (50%) and surgery (50%), and 100% were receiving parenteral nutrition; however C. parapsilosis sensu stricto was recovered primarily from patients over 50 years (69.2%) and 46.1% were receiving parenteral nutrition. CONCLUSIONS These findings show that C. orthopsilosis should be considered as human pathogenic yeast and therefore its accurate identification is important. Despite our small sample size our study suggests that a displacement of some epidemiological characteristics previously attributed to C. parapsilosis to C. orthopsilosis may be possible.

Resistance to Vancomycin, LY333328, Ciprofloxacin and Trovafloxacin of Community-Acquired and Nosocomial Strains of Enterococcus faecalis Isolated in Badajoz (Spain) with and without High-Level Resistance to Streptomycin and Gentamicin

In vitro resistance of community-acquired and nosocomial strains of Enterococcus faecalis isolated in Badajoz (Spain) were determined by a microdilution method. The isolates were identified with conventional MicroScan Pos Combo 4 I dehydrated panels. No resistance to glycopeptides was found, but LY333328 was 2–4 times more active than vancomycin. In the nosocomial strains, high-level resistance to streptomycin (HLRS) was 54.7%, and high-level resistance to gentamicin (HLRG) was 38.1%. Resistance to ciprofloxacin and trovafloxacin was 45.3 and 38.9%, respectively. In the community-acquired isolates, HLRS, HLRG, resistance to ciprofloxacin and resistance to trovafloxacin were 44.2, 17.3, 15.4 and 13.5%, respectively. Trovafloxacin was 2–4 times more active than ciprofloxacin against both groups of strains. An association between high-level resistance to aminoglycosides and resistance to fluoroquinolones was noted. The resistance to aminoglycosides did not influence the activity of vancomycin and LY333328.

Phagocytic Activity of Polymorphonuclear Leukocytes on Escherichia coli Previously Exposed to Metronidazole

A study was made of the action of different concentrations of metronidazole of the viability of Escherichia coli under aerobic and anaerobic conditions. The viability of E. coli was reduced by 60 to 99% after 24 hours of anaerobic incubation, according to the concentration of metronidazole tested. In addition, there were significant morphological changes in the bacteria. Exposure of antibiotic-induced filaments of E. coli LP 136 to phagocytosis caused the cfu/ml value to drop by 60% after 120 minutes. Under identical conditions, using the mutant strain E. coli RYC 819, which did not become filamented by metronidazole although it did present similar ultrastructural changes, this reduction reached 83%. These results may explain the therapeutic success of metronidazole in polymicrobial infections.