RESUMEN
Azoles are the most commonly used clinical antifungal therapy and also play an important role in control of plant pathogens. Intrinsic resistance to the azole class of fungicides, which target lanosterol demethylase (CYP51), is observed in many fungal species; however, the mechanisms underpinning this phenomenon are unknown. In this study, 5 azole-resistant Penicillium isolates from patients attending the UK National Aspergillosis Centre that could not be morphologically identified to species level were analyzed by genome sequencing. The genomes and CYP51 paralogue structure from these isolates were compared with those of 46 representative fungal isolates to identify to species level and examine possible mechanisms of drug resistance. Analysis of CYP51 paralogues showed that azole-resistant isolates from this study (n = 2) and from public databases (n = 6) contained a new CYP51 paralogue, CYP51D, which was associated with azole resistance in 6/8 cases and never occurred in azole-sensitive species (46/46 tested). Furthermore, one isolate from this study and an azole-resistant Aspergillus fumigatiaffinis isolate were shown to encode a CYP51A paralogue, CYP51A2. Introduction of CYP51A2 to the closely related but azole-sensitive Aspergillus fumigatus resulted in azole resistance. The identification of novel CYP51A and CYP51D paralogues in resistant fungi and the observation that resistance to azoles can be conferred by introducing a CYP51A paralogue from a resistant species into an azole-sensitive species are a potentially important new azole resistance mechanism. IMPORTANCE Azole antifungals are the main treatment for fungal disease in humans. Many species are intrinsically resistant to azoles-in other words all members of the species are resistant without prior exposure-and we do not understand why. In this study, we serendipitously discovered that many intrinsically resistant species have alternative or extra copies of the azole target gene, CYP51. Transfer of one of these genes from a resistant species to a sensitive one resulted in drug resistance, showing that the extra copies of CYP51 can confer drug resistance. Understanding how clinically important species are resistant to therapy allows us to predict whether a species could be resistant from genome sequence.
Asunto(s)
Antifúngicos/farmacología , Azoles/farmacología , Hongos/efectos de los fármacos , Esterol 14-Desmetilasa/efectos de los fármacos , Aspergilosis/microbiología , Aspergillus/efectos de los fármacos , Aspergillus/genética , Aspergillus fumigatus/efectos de los fármacos , Aspergillus fumigatus/genética , Farmacorresistencia Fúngica/genética , Proteínas Fúngicas/genética , Hongos/genética , Fungicidas Industriales/farmacología , Humanos , Pruebas de Sensibilidad Microbiana , Micosis , Esterol 14-Desmetilasa/clasificación , Esterol 14-Desmetilasa/genéticaRESUMEN
Every year, dogs are presented to veterinary clinics in the Netherlands after having been bitten by a viper. The viper is the only venomous snake native to the Netherlands. Clinical signs after an acquired viper bite can range from none (after a 'dry' bite) to very mild up to life threatening following a 'wet' bite. To prevent mortality it is important to monitor the animals for a period of time and provide adequate treatment. Clotting disorders and multiple organ failure can occur several days to a week after the viper bite, appropriate follow up is therefore important. In the Netherlands, a specific antiserum is available for veterinarians. The use of this antiserum is strongly recommended in severe cases of viper envenomation.