scholarly journals Environmental Isolates of Multi-Azole-Resistant Aspergillus spp. in Southern Italy

2018 ◽  
Vol 4 (4) ◽  
pp. 131 ◽  
Author(s):  
Laura Trovato ◽  
Guido Scalia ◽  
Maria Domina ◽  
Salvatore Oliveri
Keyword(s):  
Dna Sequencing ◽  
Southern Italy ◽  
Resistance Mechanism ◽  
Sensu Stricto ◽  
Surveillance Program ◽  
Azole Resistance ◽  
Environmental Isolates ◽  
Related Disease ◽  
First Time ◽  
Aspergillus Section

Azole resistance in Aspergillus spp. has been increasingly reported worldwide. Acquired azole resistance is probably linked to environmental exposure to fungicides used in agriculture. We collected a total of 84 soil and leaf samples from eight farms in Southern Italy. Aspergillus isolates were tested for resistance to itraconazole, posaconazole, and voriconazole by the EUCAST method. Five out of 84 samples yielded A. fumigatus isolates: four of them were itraconazole-resistant and were identified as A. fumigatus sensu stricto, three of them were posaconazole-resistant, and two were also voriconazole-resistant. All three isolates harbored the TR34/L98H resistance mechanism, which was detected by DNA sequencing of the cyp51A gene. Fifteen out of 84 samples yielded Aspergillus spp. isolates and included 11 itraconazole-resistant isolates: Aspergillus section Nigri (9) and Aspergillus section Flavi (2). Our study reports for the first time the isolation of azole-resistant A. fumigatus harboring TR34/L98H mutation from the environment of Southern Italy. The present work provides a better understanding of the magnitude of the environmental spread of azole resistance in the context of a necessary effective surveillance program to improve the management of Aspergillus-related disease.

scholarly journals Molecular Identification, Antifungal Susceptibility Testing, and Mechanisms of Azole Resistance in Aspergillus Species Received within a Surveillance Program on Antifungal Resistance in Spain

2019 ◽  
Vol 63 (9) ◽  
Author(s):  
Olga Rivero-Menendez ◽  
Juan Carlos Soto-Debran ◽  
Narda Medina ◽  
Jose Lucio ◽  
Emilia Mellado ◽  
...  
Keyword(s):  
Aspergillus Terreus ◽  
Fungal Infections ◽  
Novel Mutation ◽  
Antifungal Susceptibility ◽  
Resistance Mechanism ◽  
Sensu Stricto ◽  
Antifungal Resistance ◽  
Surveillance Program ◽  
Azole Resistance ◽  
Content Type

ABSTRACT Antifungal resistance is one of the major causes of the increasing mortality rates for fungal infections, especially for those caused by Aspergillus spp. A surveillance program was established in 2014 in the Spanish National Center for Microbiology for tracking resistance in the most prevalent Aspergillus species. A total of 273 samples were included in the study and were initially classified as susceptible or resistant according to EUCAST breakpoints. Several Aspergillus cryptic species were found within the molecularly identified isolates. Cyp51 mutations were characterized for Aspergillus fumigatus, Aspergillus terreus, and Aspergillus flavus sensu stricto strains that were classified as resistant. Three A. fumigatus sensu stricto strains carried the TR34/L98H resistance mechanism, while two harbored G54R substitution and one harbored the TR46/Y121F/T289A mechanism. Seventeen strains had no mutations in cyp51A, with ten of them resistant only to isavuconazole. Three A. terreus sensu stricto strains harbored D344N substitution in cyp51A, one of them combined with M217I, and another carried an A249G novel mutation. Itraconazole-resistant A. flavus sensu stricto strains harbored P220L and H349R alterations in cyp51A and cyp51C, respectively, that need further investigation on their implication in azole resistance.

scholarly journals Azole Resistance in Clinical and Environmental Aspergillus Isolates from the French West Indies (Martinique)

2021 ◽  
Vol 7 (5) ◽  
pp. 355
Author(s):  
Lorra Monpierre ◽  
Nicole Desbois-Nogard ◽  
Isabel Valsecchi ◽  
Marielle Bajal ◽  
Cécile Angebault ◽  
...  
Keyword(s):  
West Indies ◽  
Culture Media ◽  
Antifungal Susceptibility ◽  
Resistance Mechanisms ◽  
Clinical Samples ◽  
Azole Resistance ◽  
Environmental Isolates ◽  
French West Indies ◽  
First Time ◽  
Microdilution Broth

The emergence of azole resistant Aspergillus spp., especially Aspergillus fumigatus, has been described in several countries around the world with varying prevalence depending on the country. To our knowledge, azole resistance in Aspergillus spp. has not been reported in the West Indies yet. In this study, we investigated the antifungal susceptibility of clinical and environmental isolates of Aspergillus spp. from Martinique, and the potential resistance mechanisms associated with mutations in cyp51A gene. Overall, 208 Aspergillus isolates were recovered from clinical samples (n = 45) and environmental soil samples (n = 163). They were screened for resistance to azole drugs using selective culture media. The Minimum Inhibitory Concentrations (MIC) towards voriconazole, itraconazole, posaconazole and isavuconazole, as shown by the resistant isolates, were determined using the European Committee on Antimicrobial Susceptibility Testing (EUCAST) microdilution broth method. Eight isolates (A. fumigatus, n = 6 and A. terreus, n = 2) had high MIC for at least one azole drug. The sequencing of cyp51A gene revealed the mutations G54R and TR34/L98H in two A. fumigatus clinical isolates. Our study showed for the first time the presence of azole resistance in A. fumigatus and A. terreus isolates in the French West Indies.

scholarly journals Environmental Study of Azole-Resistant Aspergillus fumigatus and Other Aspergilli in Austria, Denmark, and Spain

2010 ◽  
Vol 54 (11) ◽  
pp. 4545-4549 ◽  
Author(s):  
Klaus Leth Mortensen ◽  
Emilia Mellado ◽  
Cornelia Lass-Flörl ◽  
Juan Luis Rodriguez-Tudela ◽  
Helle Krogh Johansen ◽  
...  
Keyword(s):  
The Netherlands ◽  
Aspergillus Fumigatus ◽  
Amphotericin B ◽  
Resistance Mechanism ◽  
Soil Samples ◽  
Environmental Study ◽  
Azole Resistance ◽  
Environmental Isolates ◽  
Intrinsic Resistance ◽  
Single Mechanism

ABSTRACT A single mechanism of azole resistance was shown to predominate in clinical and environmental Aspergillus fumigatus isolates from the Netherlands, and a link to the use of azoles in the environment was suggested. To explore the prevalence of azole-resistant A. fumigatus and other aspergilli in the environment in other European countries, we collected samples from the surroundings of hospitals in Copenhagen, Innsbruck, and Madrid, flowerbeds in an amusement park in Copenhagen, and compost bags purchased in Austria, Denmark, and Spain and screened for azole resistance using multidish agars with itraconazole, voriconazole, and posaconazole. EUCAST method E.DEF 9.1 was used to confirm azole resistance. The promoter and entire coding sequence of the cyp51A gene were sequenced to identify azole-resistant A. fumigatus isolates. A. fumigatus was recovered in 144 out of 185 samples (77.8%). Four A. fumigatus isolates from four Danish soil samples displayed elevated azole MICs (8%), and all harbored the same TR/L98H mutation of cyp51A. One A. lentulus isolate with voriconazole MIC of 4 mg/liter was detected in Spain. No azole-resistant aspergilli were detected in compost. Finally, A. terreus was present in seven samples from Austria. Multi-azole-resistant A. fumigatus is present in the environment in Denmark. The resistance mechanism is identical to that of environmental isolates in the Netherlands. No link to commercial compost could be detected. In Spain and Austria, only Aspergillus species with intrinsic resistance to either azoles or amphotericin B were found.

scholarly journals Prevalence and mechanisms of azole resistance in clinical isolates of Aspergillus section Fumigati species in a Canadian tertiary care centre, 2000 to 2013

2019 ◽  
Vol 75 (4) ◽  
pp. 849-858
Author(s):  
Maxime Parent-Michaud ◽  
Philippe J Dufresne ◽  
Eric Fournier ◽  
Benjamin Folch ◽  
Christine Martineau ◽  
...  
Keyword(s):  
Efflux Pump ◽  
Tertiary Care ◽  
Geographic Area ◽  
Resistance Mechanisms ◽  
Sensu Stricto ◽  
Azole Resistance ◽  
Intrinsic Resistance ◽  
Tertiary Care Centre ◽  
Combined Use ◽  
Aspergillus Section

Abstract Objectives Azole resistance among Aspergillus fumigatus isolates is a growing concern worldwide. Induction of mutations during azole therapy, environment-acquired mutations caused by azole fungicides and intrinsic resistance of cryptic Fumigati species all contribute to the burden of resistance. However, there is a lack of data in Canada on this emerging threat. Methods To gain insights into the magnitude and mechanisms of resistance, a 14 year collection of Aspergillus section Fumigati comprising 999 isolates from 807 patients at a Montreal hospital was screened for azole resistance, and resistance mechanisms were investigated with the combined use of genome sequencing, 3D modelling and phenotypic efflux pump assays. Results Overall azole resistance was low (4/807 patients; 0.5%). A single azole-resistant A. fumigatus sensu stricto strain, isolated from a patient with pulmonary aspergillosis, displayed efflux-pump-mediated resistance. Three patients were colonized or infected with azole-resistant cryptic Fumigati species (one Aspergillus thermomutatus, one Aspergillus lentulus and one Aspergillus turcosus). Evidence is presented that azole resistance is efflux-pump-mediated in the A. turcosus isolate, but not in the A. lentulus and A. thermomutatus isolates. Conclusions Azole resistance is rare in our geographic area and currently driven by cryptic Fumigati species. Continued surveillance of emergence of resistance is warranted.

scholarly journals Trends on Aspergillus Epidemiology—Perspectives from a National Reference Laboratory Surveillance Program

2021 ◽  
Vol 7 (1) ◽  
pp. 28
Author(s):  
Raquel Sabino ◽  
Paulo Gonçalves ◽  
Aryse Martins Melo ◽  
Daniela Simões ◽  
Mariana Oliveira ◽  
...  
Keyword(s):  
Cryptic Species ◽  
Environmental Samples ◽  
Reference Method ◽  
Sensu Stricto ◽  
Antifungal Drugs ◽  
Surveillance Program ◽  
Azole Resistance ◽  
Correct Identification ◽  
Reference Laboratory ◽  
Minimal Inhibitory Concentrations

Identification of Aspergillus to species level is important since sibling species may display variable susceptibilities to multiple antifungal drugs and also because correct identification contributes to improve the knowledge of epidemiological studies. Two retrospective laboratory studies were conducted on Aspergillus surveillance at the Portuguese National Mycology Reference Laboratory. The first, covering the period 2017–2018, aimed to study the molecular epidemiology of 256 Aspergillus isolates obtained from patients with respiratory, subcutaneous, or systemic infections and from environmental samples. The second, using our entire collection of clinical and environmental A. fumigatus isolates (N = 337), collected between 2012 and 2019, aimed to determine the frequency of azole-resistant A. fumigatus isolates. Aspergillus fumigatus sensu stricto was the most frequent species in both clinical and environmental samples. Overall, and considering all Aspergillus sections identified, a high frequency of cryptic species was detected, based on beta-tubulin or calmodulin sequencing (37% in clinical and 51% in environmental isolates). Regarding all Fumigati isolates recovered from 2012–2019, the frequency of cryptic species was 5.3% (18/337), with the identification of A. felis (complex), A. lentulus, A. udagawae, A. hiratsukae, and A. oerlinghauensis. To determine the frequency of azole resistance of A. fumigatus, isolates were screened for azole resistance using azole-agars, and 53 possible resistant isolates were tested by the CLSI microdilution reference method. Nine A. fumigatus sensu stricto and six Fumigati cryptic isolates showed high minimal inhibitory concentrations to itraconazole, voriconazole, and/or posaconazole. Real-time PCR to detect cyp51A mutations and sequencing of cyp51A gene and its promoter were performed. The overall frequency of resistance to azoles in A. fumigatus sensu stricto was 3.0%. With this retrospective analysis, we were able to detect one azole-resistant G54R mutant A. fumigatus environmental isolate, collected in 2015. The TR34/L98H mutation, linked to environmental transmission route of azole resistance, was the most frequently detected mutation (N = 4; 1.4%). Our findings underline the demand for correct identification and susceptibility testing of Aspergillus isolates.

scholarly journals Clinical and Environmental Isolates of Cryptococcus gattii from Australia That Retain Sexual Fecundity

2005 ◽  
Vol 4 (8) ◽  
pp. 1410-1419 ◽  
Author(s):  
Leona T. Campbell ◽  
James A. Fraser ◽  
Connie B. Nichols ◽  
Fred S. Dietrich ◽  
Dee Carter ◽  
...  
Keyword(s):  
Common Ancestor ◽  
Dominant Mode ◽  
Cryptococcus Gattii ◽  
Sexual Cycle ◽  
Environmental Isolates ◽  
Molecular Type ◽  
Pathogenic Yeast ◽  
Cycle Dependent ◽  
Mat Locus ◽  
First Time

ABSTRACTCryptococcus gattiiis a primary pathogenic yeast that causes disease in both animals and humans. It is closely related toCryptococcus neoformansand diverged from a common ancestor ∼40 million years ago. WhileC. gattiihas a characterized sexual cycle dependent upon a dimorphic region of the genome known as the MAT locus, mating has rarely been observed in this species. In this study, we identify for the first time clinical (both human and veterinary) and environmental isolates from Australia that retain sexual fecundity. A collection of 120 isolates from a variety of geographic locations was analyzed for molecular type, mating type, and the ability to develop mating structures when cocultured with fertile tester strains. Nine isolates produced dikaryotic filaments with paired nuclei, fused clamp connections, and basidiospores. DNA sequence analysis of three genes (URA5, the MATα-specificSXI1α gene, and the MATa-specificSXI2agene) revealed little or no variability inURA5andSXI2a, respectively. However across the 108 MATα strains sequenced, theSXI1α gene was found to exist as 11 different alleles. Phylogenetic analysis found most variation to occur in the more fertile genotypes. Although some lineages of AustralianC. gattiihave retained the ability to mate, the majority of isolates were sterile, suggesting that asexuality is the dominant mode of propagation in these populations.

NORTH AMERICAN WATER MITES OF THE FAMILY MOMONIIDAE VIETS (ACARI: ARRENUROIDEA). IV. REVISION OF SPECIES OF STYGOMOMONIA (SENSU STRICTO) SZALAY, 1943

1991 ◽  
Vol 123 (3) ◽  
pp. 501-558 ◽  
Author(s):  
Ian M. Smith
Keyword(s):  
New Species ◽  
North American ◽  
Sensu Stricto ◽  
North American Species ◽  
Water Mites ◽  
Dispersal Patterns ◽  
The Family ◽  
Three New Species ◽  
New Diagnosis ◽  
First Time

AbstractMorphological, life history, and distributional data are presented for North American species of the subgenus Stygomomonia (sensu stricto) Szalay, 1943. Adults of the seven previously recognized species are redescribed, and deutonymphs of five of these species are described for the first time. Two species, S. (s.s.) neomexicana Cook and S. (s.s.) occidentalis Cook are substantially revised on the basis of an examination of the types and extensive series of newly collected specimens. Three new species are described, S. (s.s.) californiensis on the basis of deutonymphs and adults, and S. (s.s.) imamurai and S. (s.s.) cooki on the basis of adults. A new diagnosis of the subgenus is proposed and discussed, the relationships of the various species are discussed, and a key to deutonymphs and adults of North American species is presented. New distributional data are presented for all species, and dispersal patterns from Pleistocene refugia are discussed.

Utilizing the Public GenomeTrakr Database for Foodborne Pathogen Traceback v1

2021 ◽  
Author(s):  
Ruth E Timme ◽  
Maria Sanchez ◽  
Marc Allard
Keyword(s):  
Real Time ◽  
Foodborne Pathogens ◽  
Pathogen Detection ◽  
Foodborne Pathogen ◽  
Reference Database ◽  
Surveillance Program ◽  
Environmental Isolates ◽  
The Public ◽  
The World ◽  
The Usa

This protocol outlines the all the steps necessary to become a GenomeTrakr data contributor. GenomeTrakr is an international genomic reference database of mostly food and environmental isolates from foodborne pathogens. The data and analyses are housed at the National Center for Biotechnology Information (NCBI), which is a database freely available to anyone in the world. The Pathogen Detection browser at NCBI computes daily cluster results adding the newly submitted data to the existing phylogenetic clusters of closely related genomes. Contributors to this database can see how their new isolates are related to the real-time foodborne pathogen surveillance program established in the USA and a few other countries, and at the same time adding valuable new data to the reference database. ------ Although originally published as a Chapter in Methods and Protocols, Foodborne Bacterial Pathogens, the protocol has since been adapted and split into four separate protocols all of which are contained in this collection.

Azole resistance among clinical isolates of Aspergillus fumigatus in Lima-Peru

2019 ◽  
Vol 58 (1) ◽  
pp. 54-60 ◽  
Author(s):  
Beatriz Bustamante ◽  
Luis Ricardo Illescas ◽  
Andrés Posadas ◽  
Pablo E Campos
Keyword(s):  
Aspergillus Fumigatus ◽  
Latin American ◽  
Pcr Amplification ◽  
Sensu Stricto ◽  
Clinical Isolates ◽  
Azole Resistance ◽  
Molecular Testing ◽  
Naive Patient ◽  
In Vitro Susceptibility

Abstract Azole resistance among Aspergillus fumigatus isolates, which is mainly related to mutations in the cyp51A gene, is a concern because it is rising, worldwide disseminated, and associated with treatment failure and death. Data on azole resistance of aspergillus from Latin American countries is very scarce and do not exist for Peru. Two hundred and seven Aspergillus clinical isolates collected prospectively underwent mycology and molecular testing for specie identification, and 143 isolates were confirmed as A. fumigatus sensu stricto (AFSS). All AFSS were tested for in vitro azole susceptibility, and resistant isolates underwent PCR amplification and sequencing of the whole cyp51A gene and its promoter. The in vitro susceptibility showed a minimal inhibitory concentration (MIC) range, MIC50 and MIC90 of 0.125 to >16, 0.25, and 0.5 μg/ml for itraconazole; 0.25 to 2, 0.5, and 0.5 μg/ml for voriconazole; and 0.003 to 1, 0.06, and 0.125 μg/ml for posaconazole. Three isolates (2%) showed resistance to itraconazole and exhibited different mutations of the cyp51A gene. One isolate harbored the mutation M220K, while a second one exhibited the G54 mutation plus a modification in the cyp51A gene promoter. The third isolate, from an azole naive patient, presented an integration of a 34-bp tandem repeat (TR34) in the promoter region of the gene and a substitution of leucine 98 by histidine (L98H). The three source patients had a diagnosis or suspicion of chronic pulmonary aspergillosis.

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