Fumigatus isolates from India harboring TR34/ L98H mutations in the cyp51A gene, from soil samples of paddy fields, tea gardens, cotton trees, buy BIBS39 flower pots and indoor air of hospital. Furthermore, we investigated the cross resistance of these environmental and clinical TR34/L98H A. fumigatus isolates to registered and commonly used azole fungicides in India and determined the genetic relatedness of Indian environmental and clinical A. fumigatus isolates harboring the TR34/L98H mutations and compared them with isolates from Europe and China.75), soil beneath cotton trees 20 (3/15), rice paddy fields 12.3 (12/97), air samples of hospital wards 7.6 (3/39) and from soil admixed with bird droppings 3.8 (2/52). There was no isolation of resistant A. fumigatus isolates from soil samples of public parks and gardens inside the hospital premises and red chilly fields in Tamil Nadu.Evidence for Cross-Resistance to Triazole Antifungal DrugsAll the 44 ITC+ A. fumigatus isolates from the environment showed reduced susceptibility to azoles. The geometric mean (GM) MIC of itraconazole (GM, 16 mg/L) was the highest, followed by voriconazole (GM, 8.7 mg/L), and posaconazole (GM, 1.03 mg/L). All the antifungal drugs tested showed reduced efficacy against all the ITC+ A. fumigatus isolates (Table 2), consistent with cross-resistance of these isolates to the tested azoles. Among the triazoles, the MIC difference between wild type and TR34/L98H isolates were the highest for itraconazole (r = 0.96) followed by voriconazole (r = 0.91) and posaconazole (r = 0.72). Of the10 fungicides, 7 showed dissimilarity between the MICs with greatest differences found for bromuconazole, difenoconazole, tebuconazole (r = 0.96 each) followed by hexaconazole (r = 0.95), epoxiconazole (r = 0.92), metconazole (r = 0.89) and lowest for cyproconazole (r = 0.22) (Table 2).Evidence for Clonal Spread of a Single Triazole-Resistant A. fumigatus GenotypeOur genotype analyses identified that all of the 44 ITC+ A. fumigatus isolates from India exhibited the same TR34/L98H genotype at the cyp51A gene. Furthermore, these strains had the same allele across all nine examined microsatellite loci (Fig. 2). In contrast to the genetic uniformity of azole-resistant strains from India, the azole-susceptible isolates from both patients and environments in India were genetically very diverse. Indeed, all nine loci were highly polymorphic in populations of azolesusceptible isolates from both clinical and environmental samples.Results Isolation of Environmental Strains of A. fumigatusOf the 486 environmental samples tested, 201 (41.4 ) showed the presence of A. fumigatus in all types of substrates tested except nursery plants soil and decayed wood inside tree trunk hollows. The data of state-wise distribution and prevalence of azole resistant A. fumigatus in soil and air samples is presented in Table 1 and Figure 1. Of the 201 A. fumigatus positive samples, 630 individual A. fumigatus colonies were obtained from I-BRD9 Sabourauds dextrose agar (SDA) plates. The count of A. fumigatus on primary SDA plate ranged from one colony to confluent growth. Besides A. niger, A. flavus, A. terreus, other molds such as mucorales, and Penicillium species were also observed in soil samples. Out of 630 A. fumigatus colonies tested, 44 (7 ) isolates originating from 24 samples grew on SDA plates containing 4 mg/L itraconazole. Among these 44 itraconazole-resistant (ITC+) isolates, 15 were obtained from different potted pl.Fumigatus isolates from India harboring TR34/ L98H mutations in the cyp51A gene, from soil samples of paddy fields, tea gardens, cotton trees, flower pots and indoor air of hospital. Furthermore, we investigated the cross resistance of these environmental and clinical TR34/L98H A. fumigatus isolates to registered and commonly used azole fungicides in India and determined the genetic relatedness of Indian environmental and clinical A. fumigatus isolates harboring the TR34/L98H mutations and compared them with isolates from Europe and China.75), soil beneath cotton trees 20 (3/15), rice paddy fields 12.3 (12/97), air samples of hospital wards 7.6 (3/39) and from soil admixed with bird droppings 3.8 (2/52). There was no isolation of resistant A. fumigatus isolates from soil samples of public parks and gardens inside the hospital premises and red chilly fields in Tamil Nadu.Evidence for Cross-Resistance to Triazole Antifungal DrugsAll the 44 ITC+ A. fumigatus isolates from the environment showed reduced susceptibility to azoles. The geometric mean (GM) MIC of itraconazole (GM, 16 mg/L) was the highest, followed by voriconazole (GM, 8.7 mg/L), and posaconazole (GM, 1.03 mg/L). All the antifungal drugs tested showed reduced efficacy against all the ITC+ A. fumigatus isolates (Table 2), consistent with cross-resistance of these isolates to the tested azoles. Among the triazoles, the MIC difference between wild type and TR34/L98H isolates were the highest for itraconazole (r = 0.96) followed by voriconazole (r = 0.91) and posaconazole (r = 0.72). Of the10 fungicides, 7 showed dissimilarity between the MICs with greatest differences found for bromuconazole, difenoconazole, tebuconazole (r = 0.96 each) followed by hexaconazole (r = 0.95), epoxiconazole (r = 0.92), metconazole (r = 0.89) and lowest for cyproconazole (r = 0.22) (Table 2).Evidence for Clonal Spread of a Single Triazole-Resistant A. fumigatus GenotypeOur genotype analyses identified that all of the 44 ITC+ A. fumigatus isolates from India exhibited the same TR34/L98H genotype at the cyp51A gene. Furthermore, these strains had the same allele across all nine examined microsatellite loci (Fig. 2). In contrast to the genetic uniformity of azole-resistant strains from India, the azole-susceptible isolates from both patients and environments in India were genetically very diverse. Indeed, all nine loci were highly polymorphic in populations of azolesusceptible isolates from both clinical and environmental samples.Results Isolation of Environmental Strains of A. fumigatusOf the 486 environmental samples tested, 201 (41.4 ) showed the presence of A. fumigatus in all types of substrates tested except nursery plants soil and decayed wood inside tree trunk hollows. The data of state-wise distribution and prevalence of azole resistant A. fumigatus in soil and air samples is presented in Table 1 and Figure 1. Of the 201 A. fumigatus positive samples, 630 individual A. fumigatus colonies were obtained from Sabourauds dextrose agar (SDA) plates. The count of A. fumigatus on primary SDA plate ranged from one colony to confluent growth. Besides A. niger, A. flavus, A. terreus, other molds such as mucorales, and Penicillium species were also observed in soil samples. Out of 630 A. fumigatus colonies tested, 44 (7 ) isolates originating from 24 samples grew on SDA plates containing 4 mg/L itraconazole. Among these 44 itraconazole-resistant (ITC+) isolates, 15 were obtained from different potted pl.