[30], and 48 h for Mucor.[12] Since Syncephalastrum, Lichtheimia and Apophysomyces revealed inadequate growth in the control well after 48 h, therefore, MIC readings were taken after 72 h. MIC end points for all the drugs except echinocandins were defined as the lowest concentration that produced complete inhibition of growth viz-à-viz the hyphal growth in
the control well. Minimum effective concentration of echinocandins were defined as the lowest drug concentrations that allowed the growth of small, rounded, degenerated colonies viz-à-viz the hyphal growth in the control well. Clinical breakpoints for mucorales are not yet published, therefore, the break points referred by Almyroudis et al. [12] for testing 217 clinical isolates of zygomycetes were used for analysis, viz, AMB ≤ 1 μg ml−1; ITC ≤ 0.5 μg ml−1; VRC ≤ 2 μg ml−1; POS ≤ 0.5 μg ml−1; BMN 673 mouse FLU ≤ 32 μg ml−1 and CAS ≤ 2 μg ml−1. For ISA, recently established ECVs of ≤1 μg ml−1 for Aspergillus species were used.[32] Susceptibility to POS and AMB was also determined by Etest method (AB Biodisk, BioMérieux, Marcy l’Etoile, France).[33] The inoculum were prepared as above to obtain a density of 0.2–2.5 × 105 cells ml−1 Selleckchem MG-132 measured by spectrophotometer. A swab was dipped into the suspension and streaked across the surface
of antibiotic medium 3 (Difco, New Jersey, USA) agar plates for testing AMB and on RPMI agar plates with 2% glucose for POS. The plates were incubated at 35 °C and the lowest drug concentration at which the border of the elliptical inhibition zone intercepted the scale on the antifungal strip was recorded at 24 h for Rhizopus spp. and at 48 h for the other species. Statistical analyses were performed with spss version 20.0 (SPSS, Chicago, IL, USA). MIC values of CLSI and Etest methods were assessed, using the Student’s t-test (paired sample). Categorical agreement between the MICs obtained by the CLSI microdilution and Etest method was calculated for AMB and POS for which above described
breakpoints were used for analysis. Of the 71 patients with mucormycosis, 39 were diagnosed as pulmonary, 15 as rhino-cerebral, 13 as cutaneous/subcutaneous and 4 as disseminated. Treatment and outcome records were available for 54 patients science (28 pulmonary, 12 sinus infection with or without brain invasion and or ocular involvement, 10 cutaneous/subcutaneous and 4 disseminated). Of these, the disease was fatal in 28 cases (51.8%), which included 12 (42.8%) cases of pulmonary, 11 (39.2%) of rhino-cerebral, 4 cases of disseminated and 1 of cutaneous mucormycosis. Overall, the commonest underlying condition in mucormycosis was uncontrolled diabetes mellitus (47%), followed by haematological malignancies (24%), chronic obstructive pulmonary disease (COPD) with long-term steroid use (20%) and trauma (9%).