Allodiploid hybrid species, Aspergillus latus, belonging to section Nidulantes, is a hybrid of A. spinulosporus and an unknown species closely related to A. quadrilineatus and A. sublatus. This hybrid has often been misidentified as the species in section Nidulantes, such as A. nidulans, A. spinulosporus, A. sublatus, or other cryptic species. Aspergillus latus has not been reported in Japan as well as Asia so far. In this study, we screened 23 clinical strains identified as A. spinulosporus isolated in Japan from 2012 to 2023 and found seven A. latus strains. To characterize the A. latus strains, we conducted comprehensive phenotyping including morphological observation, whole genome sequences, and phylogenetic analysis based on calmodulin (CaM) gene. In addition, we conducted antifungal susceptibility testing for A. latus strains. As a result, the morphological characters of A. latus were more similar to those of A. spinulosporus compared to A. sublatus. However, the ascospore of A. latus differed from that of A. spinulosporus. Phylogenetic analysis revealed that different CaM alleles from the same isolate clustered separately with A. spinulosporus and A. sublatus, consistent with its hybrid origin. Furthermore, A. latus strains showed reduced susceptibility to caspofungin and amphotericin B compared to A. spinulosporus, while they were susceptible to azoles. Our results suggest that A. latus has been a causative pathogen of aspergillosis in Japan since 2013.
- MeSH
- Antifungal Agents pharmacology MeSH
- Aspergillus * genetics classification isolation & purification drug effects MeSH
- Aspergillosis * microbiology epidemiology MeSH
- Phylogeny MeSH
- Calmodulin genetics MeSH
- Humans MeSH
- Microbial Sensitivity Tests MeSH
- Whole Genome Sequencing MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Geographicals
- Japan MeSH
UNLABELLED: The Aspergillus genus encompasses a diverse array of species, some of which are opportunistic pathogens. Traditionally, human aspergillosis has primarily been linked to a few Aspergillus species, predominantly A. fumigatus. Changes in epidemiology and advancements in molecular techniques have brought attention to less common and previously unrecognized pathogenic cryptic species. Despite the taxonomic recognition of many cryptic species in section Terrei, their virulence potential and clinical implications, compared to A. terreus sensu stricto, remain poorly understood. Hence, the current study utilized the alternative in vivo model Galleria mellonella to evaluate the virulence potential of 19 accepted Aspergillus species in section Terrei, classified into three series (major phylogenetic clades): Terrei, Nivei, and Ambigui. Analyzing the median survival rates of infected larvae of all species in each series revealed that series Ambigui has a significantly lower virulence compared to series Terrei and Nivei. Taking a closer look at series Terrei and Nivei revealed a trend of survival within each clade, dividing the species into two groups: highly virulent (up to 72 h survival) and less virulent (up to 144 h survival). Histological observation, considering fungal distribution and filamentation, further supported this assessment, revealing increased distribution and hyphal formation in virulent species. Additionally, the susceptibility profile of conventional antifungals was determined, revealing an increased azole minimum inhibitory concentration for some tested cryptic species such as A. niveus and A. iranicus. Our results highlight the importance of cryptic species identification, as they can exhibit different levels of virulence and show reduced antifungal susceptibility. IMPORTANCE: With changing fungal epidemiology and an increasingly vulnerable population, cryptic Aspergillus species are emerging as human pathogens. Their diversity and clinical relevance remain underexplored, with some species showing reduced antifungal susceptibility and higher virulence, highlighting the need for better preparedness in clinical practice. Using the Galleria mellonella model, we assessed the virulence of Aspergillus species of section Terrei, including cryptic and non-cryptic species, across three series Terrei, Nivei, and Ambigui. The results revealed significant virulence variation among the series, with some cryptic species displaying high virulence. Histological analysis confirmed increased hyphal formation and fungal spread in the more virulent species. Additionally, elevated azole minimum inhibitory concentrations were also observed in certain cryptic species. This study presents novel insights into the pathogenicity of Aspergillus section Terrei, emphasizing the critical importance of accurately identifying cryptic species due to their diverse virulence potential and antifungal resistance, which may have substantial clinical implications.
- MeSH
- Antifungal Agents pharmacology MeSH
- Aspergillus * pathogenicity classification drug effects genetics MeSH
- Aspergillosis * microbiology MeSH
- Phylogeny MeSH
- Larva microbiology MeSH
- Humans MeSH
- Disease Models, Animal MeSH
- Moths * microbiology MeSH
- Virulence MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
Aspergillus section Nidulantes encompasses almost 80 homothallic and anamorphic species, mostly isolated from soil, plant material, or the indoor environment. Some species are clinically relevant or produce mycotoxins. This study reevaluated the species boundaries within several clades of section Nidulantes. Five data sets were assembled, each containing presumptive new species and their closest relatives, and phylogenetic and phenotypic analyses were performed. We tested the hypotheses that the newly isolated or reexamined strains constitute separate species (splitting approach) or should be treated as part of broadly defined species (lumping approach). Four DNA sequence loci were amplified, internal transcribed spacer (ITS) and large subunit (LSU) regions of the rDNA and partial sequences of the β-tubulin (benA), calmodulin (CaM), and RNA polymerase II second largest subunit (RPB2) genes. The latter three loci were used for the phylogenetic analysis and served as input for single-locus (GMYC, bGMYC, PTP, and bPTP) and multilocus (STACEY and BP&P) species delimitation analyses. The phenotypic analysis comprised macro- and micromorphology (including scanning electron microscopy) and comparison of cardinal growth temperatures. The phylogenetic analysis supported the splitting hypothesis in all cases, and based on the combined approach, we propose six new species, four that are homothallic and two anamorphic. Four new species were isolated from the indoor environment (Jamaica, Trinidad and Tobago, USA), one originated from soil (Australia), and one from a kangaroo rat cheek pouch (USA).
- MeSH
- Aspergillus * classification cytology genetics isolation & purification MeSH
- Phylogeny MeSH
- Genes, Fungal MeSH
- Multilocus Sequence Typing MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Geographicals
- Australia MeSH
- Jamaica MeSH
- United States MeSH
- Trinidad and Tobago MeSH
Invasive fungal disease represents one of the severe complications in haematopoietic stem cell transplant recipients. We describe a case of a patient treated for relapse of chronic lymphoblastic leukaemia 6 years after HSCT. The patient was treated for invasive pulmonary aspergillosis but died 3 months later from multiple organ failures consisting of haemorrhagic necrotizing fungal pneumonia, refractory chronic hepatic graft versus host disease and cytomegalovirus hepatitis. Autopsy samples revealed histopathological evidence of fungal hyphae and an unusual Aspergillus nidulans-like species was isolated in pure culture. More precise identification was achieved by using scanning electron microscopy of ascospores and sequencing of calmodulin gene, and the isolate was subsequently re-identified as A. sublatus (section Nidulantes) and showed good in vitro susceptibility against all classes of antifungals. Commonly used ITS rDNA region and β-tubulin gene fail to discriminate A. sublatus from related pathogenic species, especially A. quadrilineatus and A. nidulans. Although this is the first case of proven IPA attributed to A. sublatus, we demonstrated that at least some previously reported infections due to A. quadrilineatus were probably caused by this cryptic species.
- MeSH
- Antifungal Agents administration & dosage MeSH
- Aspergillus classification cytology genetics isolation & purification MeSH
- Leukemia, Lymphocytic, Chronic, B-Cell complications MeSH
- Cytomegalovirus Infections complications diagnosis MeSH
- DNA, Fungal chemistry genetics MeSH
- Fatal Outcome MeSH
- Phylogeny MeSH
- Invasive Pulmonary Aspergillosis diagnosis drug therapy microbiology pathology MeSH
- Calmodulin genetics MeSH
- Middle Aged MeSH
- Humans MeSH
- DNA, Ribosomal Spacer chemistry genetics MeSH
- Microbial Sensitivity Tests MeSH
- Microscopy, Electron, Scanning MeSH
- Graft vs Host Disease complications diagnosis MeSH
- Transplant Recipients MeSH
- Sequence Analysis, DNA MeSH
- Cluster Analysis MeSH
- Hematopoietic Stem Cell Transplantation adverse effects MeSH
- Tubulin genetics MeSH
- Hepatitis, Viral, Human complications diagnosis MeSH
- Check Tag
- Middle Aged MeSH
- Humans MeSH
- Male MeSH
- Publication type
- Journal Article MeSH
- Case Reports MeSH
Aspergillus candidus is a species frequently isolated from stored grain, food, indoor environments, soil and occasionally also from clinical material. Recent bioprospecting studies highlighted the potential of using A. candidus and its relatives in various industrial sectors as a result of their significant production of enzymes and bioactive compounds. A high genetic variability was observed among A. candidus isolates originating from various European countries and the USA, that were mostly isolated from indoor environments, caves and clinical material. The A. candidus sensu lato isolates were characterized by DNA sequencing of four genetic loci, and agreement between molecular species delimitation results, morphological characters and exometabolite spectra were studied. Classical phylogenetic methods (maximum likelihood, Bayesian inference) and species delimitation methods based on the multispecies coalescent model supported recognition of up to three species in A. candidus sensu lato. After evaluation of phenotypic data, a broader species concept was adopted, and only one new species, Aspergillus dobrogensis, was proposed. This species is represented by 22 strains originating from seven countries (ex-type strain CCF 4651T=NRRL 62821T=IBT 32697T=CBS 143370T) and its differentiation from A. candidus is relevant for bioprospecting studies because these species have different exometabolite profiles. Evaluation of the antifungal susceptibility of section Candidi members to six antifungals using the reference EUCAST method showed that all species have low minimum inhibitory concentrations for all tested antifungals. These results suggest applicability of a wide spectrum of antifungal agents for treatment of infections caused by species from section Candidi.
OBJECTIVES: A prospective international multicentre surveillance study was conducted to investigate the prevalence and amphotericin B susceptibility of Aspergillus terreus species complex infections. METHODS: A total of 370 cases from 21 countries were evaluated. RESULTS: The overall prevalence of A. terreus species complex among the investigated patients with mould-positive cultures was 5.2% (370/7116). Amphotericin B MICs ranged from 0.125 to 32 mg/L, (median 8 mg/L). CONCLUSIONS: Aspergillus terreus species complex infections cause a wide spectrum of aspergillosis and the majority of cryptic species display high amphotericin B MICs.
- MeSH
- Amphotericin B pharmacology MeSH
- Antifungal Agents pharmacology MeSH
- Aspergillus classification drug effects isolation & purification MeSH
- Aspergillosis epidemiology microbiology MeSH
- Epidemiological Monitoring MeSH
- Humans MeSH
- Microbial Sensitivity Tests MeSH
- Prevalence MeSH
- Prospective Studies MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Multicenter Study MeSH
- Geographicals
- Europe epidemiology MeSH
A wide array of PCR tests has been developed to aid the diagnosis of invasive aspergillosis (IA), providing technical diversity but limiting standardisation and acceptance. Methodological recommendations for testing blood samples using PCR exist, based on achieving optimal assay sensitivity to help exclude IA. Conversely, when testing more invasive samples (BAL, biopsy, CSF) emphasis is placed on confirming disease, so analytical specificity is paramount. This multicenter study examined the analytical specificity of PCR methods for detecting IA by blind testing a panel of DNA extracted from a various fungal species to explore the range of Aspergillus species that could be detected, but also potential cross reactivity with other fungal species. Positivity rates were calculated and regression analysis was performed to determine any associations between technical specifications and performance. The accuracy of Aspergillus genus specific assays was 71.8%, significantly greater (P < .0001) than assays specific for individual Aspergillus species (47.2%). For genus specific assays the most often missed species were A. lentulus (25.0%), A. versicolor (24.1%), A. terreus (16.1%), A. flavus (15.2%), A. niger (13.4%), and A. fumigatus (6.2%). There was a significant positive association between accuracy and using an Aspergillus genus PCR assay targeting the rRNA genes (P = .0011). Conversely, there was a significant association between rRNA PCR targets and false positivity (P = .0032). To conclude current Aspergillus PCR assays are better suited for detecting A. fumigatus, with inferior detection of most other Aspergillus species. The use of an Aspergillus genus specific PCR assay targeting the rRNA genes is preferential.
- MeSH
- Aspergillus classification genetics isolation & purification MeSH
- Molecular Diagnostic Techniques methods MeSH
- Invasive Pulmonary Aspergillosis diagnosis MeSH
- Humans MeSH
- Polymerase Chain Reaction methods MeSH
- Sensitivity and Specificity MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Multicenter Study MeSH
Aspergily jsou ubikvitní houby, které u vnímavých jedinců mohou způsobit závažná onemocnění. Nejčastěji infikovaným orgánem jsou plíce. Závažnost onemocnění je závislá na stupni invaze houby do plicní tkáně, ta stoupá úměrně s rozvojem imunodeficitu pacienta. Invazivitu procesu prokáže jednoznačně pouze histologické vyšetření vzorku infikované plicní tkáně. Klasické mykologické metody detekují mikroskopickými nebo kultivačními technikami přítomnost houby ve vyšetřovaných vzorcích z dýchacích cest. Dále je možné prokázat v séru nebo bronchoalveolární tekutině specifický aspergilový antigen galaktomanan a nespecifický beta-D-glukan. Detekce protilátek proti aspergilům má přínos pouze u chronických forem onemocnění. V článku se autorka zabývá přínosem jednotlivých mykologických vyšetřovacích metod v diagnostice různých forem plicní aspergilózy.
Aspergillus are ubiquitous fungi that can cause serious illnesses in susceptible individuals. The most commonly infected organ is the lungs. The severity of the disease depends on the degree of the invasion of the lung tissue by fungi, which rises proportionally with the development of immunodeficiency. The only way to clearly determine the degree to which the lungs have been invaded is to carry out a histological examination of a sample of the infected tissue. Conventional mycological methods can detect the presence of the fungus in samples from a patient's airways by using microscopy or culture techniques. Furthermore, it is possible to determine the presence of the specific Aspergillus antigen, galactomannan, and of non-specific beta-D-glucan in the serum or bronchoalveolar lavage fluid. The detection of antibodies against Aspergillus is only relevant for chronic forms of the disease. This article discusses the benefits of different mycological examination methods in the diagnosis of various forms of pulmonary aspergillosis.
- Keywords
- galaktomanan, lateral flow device,
- MeSH
- Aspergillus classification pathogenicity MeSH
- beta-Glucans analysis MeSH
- Chromatography, Affinity methods MeSH
- Culture Techniques MeSH
- Humans MeSH
- Lipopolysaccharides blood MeSH
- Mycological Typing Techniques MeSH
- Lung anatomy & histology MeSH
- Pulmonary Aspergillosis * diagnosis pathology MeSH
- Sensitivity and Specificity MeSH
- Check Tag
- Humans MeSH
- Publication type
- Overall MeSH
The aim of this study was to experimentally test chromogenic media and reagents for a rapid diagnosis of toxigenic microfungi Aspergillus flavus, A. carbonarius, A. ochraceus, Penicillium expansum, P. verrucosum, and P. citrinum isolated from foodstuffs. Toxigenic microfungi of Aspergillus and Penicillium species are considered to be of major relevance for human and animal health. The presence of these toxigenic microfungi in foodstuffs and feedstuffs raises a potential risk of mycotoxins (for example presence of aflatoxins, cyclopiazonic acid, ochratoxin A, patulin or citrinin). In order to reduce the risks for consumers’ health, it is necessary to set rapid methods for toxigenic microfungi isolated from foodstuffs and also feedstuffs. Application of chromogenic media and reagents presents one of the possibilities. The selected strains of toxigenic microfungi, producing mycotoxins, were tested on chromogenic media. The chromogenic media and reagents in our study are suitable for a microbiological and mycological laboratory. The benefits of chromogenic media and reagents include a rapid diagnosis within 48–72 h, simple preparation, easy availability and relatively low prices.
- MeSH
- Food Analysis MeSH
- Aspergillus isolation & purification classification pathogenicity MeSH
- Food Safety MeSH
- Chromogenic Compounds MeSH
- Indicators and Reagents MeSH
- Animal Feed MeSH
- Culture Media * classification MeSH
- Microbiological Techniques * methods MeSH
- Mitosporic Fungi * isolation & purification classification metabolism pathogenicity MeSH
- Mycotoxins biosynthesis isolation & purification classification adverse effects MeSH
- Penicillium isolation & purification classification pathogenicity MeSH
- Food Microbiology MeSH
- Publication type
- Research Support, Non-U.S. Gov't MeSH
Aspergillus section Flavipedes contains species found worldwide in soils and rhizospheres, indoor and cave environments, as endophytes, food contaminants and occasionally as human pathogens. They produce many extensively studied bioactive secondary metabolites and biotechnologically relevant enzymes. The taxa were revised based on phylogenetic analysis of sequences from four loci (β-tubulin, calmodulin, RPB2, ITS rDNA), two PCR fingerprinting methods, micro- and macromorphology and physiology. Section Flavipedes includes three known and seven new species: A. ardalensis, A. frequens, A. luppii, A. mangaliensis, A. movilensis, A. polyporicola and A. spelaeus. The name A. neoflavipes was proposed for Fennellia flavipes a distinct species from its supposed asexual state A. flavipes. Aspergillus iizukae, A. frequens and A. mangaliensis are the most common and widely distributed species, whereas A. flavipes s. str. is rare. A dichotomous key based on the combination of morphology and physiology is provided for all recognized species. Aspergillus section Jani is established to contain A. janus and A. brevijanus, species previously classified as members of sect. Versicolores, Terrei or Flavipedes. This new section is strongly supported by phylogenetic data and morphology. Section Jani species produce three types of conidiophores and conidia, and colonies have green and white sectors making them distinctive. Accessory conidia found in pathogenic A. terreus were found in all members of sects. Flavipedes and Jani. Our data indicated that A. frequens is a clinically relevant and produces accessory conidia during infection.
- MeSH
- Aspergillus classification genetics growth & development isolation & purification MeSH
- Aspergillosis MeSH
- DNA, Fungal genetics MeSH
- Fungal Proteins genetics MeSH
- Phylogeny MeSH
- Humans MeSH
- Molecular Sequence Data MeSH
- Food Microbiology MeSH
- Soil Microbiology MeSH
- Spores, Fungal classification genetics growth & development isolation & purification MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
