Table 1.
Susceptibility results obtained by CLSI/EUCAST microdilution (azoles) or by Etest (echinocandins) for four sequential isolates obtained from a CGD patient over a 127 week period.
Figure 1.
In vivo susceptibility of sequential A. fumigatus isolates from a CGD patient failing azole therapy.
Susceptibility is shown as survival curve (a and d), as fungal CFU kidney burden (b and e) and as Aspergillus DNA load in kidney tissue. The early susceptible isolate (isolate no 2) is shown in fig. a, b and c and the late resistant isolate (isolate no 4) in fig. c, d and e. AND: anidulafungin, POS: posaconazole, AND+POS: combination therapy of anidulafungin and posaconazole, d4: day 4, d8: day 8, * P<0.05 compared to control.
Figure 2.
CYP51A mRNA levels in the 4 sequential A. fumigatus isolates and in a control wild type isolate AZN 8196 (WT) and a tri-azole resistant control isolate with over-expression of the CYP51A gene due to the L98H mutation and promoter tandem repeat.
Figure 3.
In vivo virulence of sequential isolates of A. fumigatus in immunosuppressed mice.
Survival of mice in groups of six challenged with each of the four sequential isolates (1–4) or a control isolate (NCPF2109), respectively, is shown. The survival data was combined per isolate, thus each survival curve displays the mortality of six mice challenged with the indicated isolate, two of which were challenged with a high inoculum (5x104 CFU/ml), two with an intermediate inoculum (104 CFU/ml) and two with a low inoculum (2×103 CFU/ml).
Figure 4.
Growth rate of the 4 sequential isolates expressed as radial growth rate on solid V8 agar (a) and as optical densities in fluent medium (b).
The laboratory strain NCP2109 is included as unrelated comparator. In the radial growth experiment (a) the growth rate is expressed in mean diameter (mm) and Kr (mm/hour) after 24–72 hours of two separate experiments performed in triplicate. In the kinetic evaluation of growth over time in liquid medium (b) growth is expressed as optical densities measured every 10 min.