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1.
Appl Environ Microbiol ; 89(6): e0007823, 2023 06 28.
Article in English | MEDLINE | ID: mdl-37255465

ABSTRACT

The preharvest maize mycobiome may be crucial in defining the health of the crop in terms of potential disease burden and mycotoxins. We investigated the preharvest maize mycobiome structure, including the influence of weather patterns, in terms of rainfall intensity, on its composition. In addition, we investigated correlation of genera Fusarium and Aspergillus with maize fumonisin-B1 and aflatoxin. Forty maize fields from selected districts in the wetter northern (N) and drier southern (S) agroecological zones of Zambia were sampled twice over two seasons (1 and 2). The defined weather variables over the two seasons were low rainfall with dry spell (S1), low rainfall (S2), and high rainfall (N1 and N2). High-throughput DNA amplicon sequencing of internal transcribed spacer 1 (ITS1) was used to determine the mycobiome structure and the composition in relation to rainfall patterns. We detected 61 genera, with Fusarium and previously unreported Sarocladium in Zambia to have the highest frequency of detection on the maize. There was a significant difference in fungal genera composition between S1 and S2 but no difference between N1 and N2. The weather pattern with dry spell, S1, had a strong proliferation of Meyerozyma and xerophiles Penicillium, Kodamaea, and Aspergillus. The four genera drove the difference in composition between S1 and S2 and the significantly higher fungal diversity in S1 compared to N2. Of the mycotoxin-important fungi, dry conditions (S1) were a key driver for proliferation of Aspergillus, while Fusarium proliferation occurred irrespective of weather patterns. The relative abundance of Aspergillus and Fusarium resonated with maize aflatoxin and fumonisin-B1 levels, respectively. IMPORTANCE Fungi contaminate various crops worldwide. Maize, an important human staple and livestock cereal, is susceptible to contamination with fungi in the field. Fungi are drivers of plant disease and can compromise yield. Some species of fungi are known to produce chemical compounds (mycotoxins), which are cancer-causing agents in humans and impair livestock productivity. It is important to understand the spectrum of fungi on maize and how weather conditions can impact their abundance. This is because the abundance of fungi in the field can have a bearing on the health of the crop as well as potential for mycotoxins contamination. By understanding the spectrum of the preharvest fungi, it becomes possible to know the key fungi adapted to the maize and subsequently the potential for crop disease as well as mycotoxins contamination. The influence of weather conditions on the spectrum of preharvest fungi on maize has not been fully explored.


Subject(s)
Aflatoxins , Fusarium , Mycobiome , Mycotoxins , Humans , Mycotoxins/analysis , Zea mays/chemistry , Zambia , Aspergillus , Food Contamination/analysis
2.
Vet Pathol ; 57(1): 139-146, 2020 01.
Article in English | MEDLINE | ID: mdl-31662046

ABSTRACT

With the aim of evaluating the presence of Fusarium spp. in sea turtles with and without lesions and assessing the risk factors favoring colonization and/or infection, 74 loggerhead sea turtles (Caretta caretta) admitted to rescue and rehabilitation clinics in Italy were analyzed. The study compared 31 individuals with no apparent macroscopic lesions and 43 individuals with macroscopic lesions. Shell and skin samples were analyzed using Calcofluor white with 10% potassium hydroxide, standard histopathological examination, and fungal cultures. Fusarium spp. were isolated more frequently from animals with superficial lesions (39%) than from those with no macroscopic lesions (16%). Isolates from animals with superficial lesions were Fusarium solani species complex (FSSC) lineages haplotypes 9, 12, and 27 (unnamed lineages), FSSC-2 (Fusarium keratoplasticum), Fusarium oxysporum (27%), and Fusarium brachygibbosum (3%). In contrast, only F. solani haplotypes 9 and 12 were isolated from animals with no macroscopic lesions. The presence of lesions was identified as a risk factor for the occurrence of Fusarium spp. Of the 74 animals, only 7 (9.5%) scored positive on microscopic examination with Calcofluor, and histological examination of those 7 animals revealed necrosis, inflammatory cells, and fungal hyphae in the carapace and skin. The results of this study suggest that fusariosis should be included in the differential diagnosis of shell and skin lesions in sea turtles. Direct examination using Calcofluor and potassium hydroxide was not useful to diagnose the infection. Histopathological examination and fungal culture should be performed to ensure correct treatment and infection control.


Subject(s)
Fusariosis/veterinary , Fusarium/isolation & purification , Necrosis/veterinary , Turtles/microbiology , Animal Shells/microbiology , Animal Shells/pathology , Animals , Female , Fusariosis/microbiology , Fusariosis/pathology , Hyphae , Italy , Male , Necrosis/microbiology , Necrosis/pathology , Skin/microbiology , Skin/pathology
3.
BMC Genomics ; 20(1): 570, 2019 Jul 10.
Article in English | MEDLINE | ID: mdl-31291889

ABSTRACT

BACKGROUND: Cucumber Fusarium wilt, caused by Fusarium oxysporum f. sp. cucumerinum (Foc), is one of the most notorious diseases in cucumber production. Our previous research showed the virulence of Foc significantly increases over consecutive rounds of infection in a resistant cultivar. To understand the virulence variation of Foc under host pressure, the mildly virulent strain foc-3b (WT) and its virulence-enhanced variant Ra-4 (InVir) were selected and their transcriptome profiles in infected cucumber roots were analyzed at 24 h after inoculation (hai) and 120 hai. RESULTS: A series of differentially expressed genes (DEGs) potentially involved in fungal pathogenicity and pathogenicity variation were identified and prove mainly involved in metabolic, transport, oxidation-reduction, cell wall degradation, macromolecules modification, and stress and defense. Among these DEGs, 190 up- and 360 down-regulated genes were expressed in both strains, indicating their importance in Foc infection. Besides, 286 and 366 DEGs showed up-regulated expression, while 492 and 214 showed down-regulated expression in InVir at 24 and 120 hai, respectively. These DEGs may be involved in increased virulence. Notably, transposases were more active in InVir than WT, indicating transposons may contribute to adaptive evolution. CONCLUSIONS: By a comparative transcriptome analysis of the mildly and highly virulent strains of Foc during infection of cucumber, a series of DEGs were identified that may be associated with virulence. Hence, this study provides new insight into the transcriptomic profile underlying pathogenicity and virulence differentiation of Foc.


Subject(s)
Cucumis sativus/microbiology , Fusarium/genetics , Fusarium/pathogenicity , Gene Expression Profiling , Adaptation, Physiological/genetics , Fusarium/physiology , Gene Regulatory Networks , Plant Roots/microbiology , Species Specificity , Virulence/genetics
4.
Med Mycol ; 56(suppl_1): 144-152, 2018 Apr 01.
Article in English | MEDLINE | ID: mdl-29538734

ABSTRACT

The Fusarium solani species complex (FSSC) is the most common group of fusaria associated with superficial and life-threatening infections in humans. Here we formally introduce Fusarium metavorans sp. nov., widely known as FSSC6 (Fusarium solani species complex lineage 6), one of the most frequent agents of human opportunistic infections. The species is described with multilocus molecular data including sequences of internal transcribed spacer region (ITS), portions of the translation elongation factor 1-a gene (TEF1), and the partial RNA polymerase II gene (rPB2). A phylogenetic approach was used to evaluate species delimitation. Topologies of the trees were concordant. Phylogenetic analyses suggest that the FSSC consists of three major clades encompassing a large number of phylogenetic species; Fusarium metavorans corresponds to phylogenetic species 6 within FSSC clade 3. The species has a global distribution and a wide ecological amplitude, also including strains from soil and agents of opportunistic plant disease; it was also isolated from the gut of the wood-boring cerambycid beetle Anoplophora glabripennis.


Subject(s)
Fusariosis/microbiology , Fusarium/classification , Fusarium/physiology , Phylogeny , Antifungal Agents/pharmacology , DNA, Fungal , DNA, Ribosomal Spacer/genetics , Fusarium/drug effects , Microbial Sensitivity Tests , Microbial Viability/drug effects , Multilocus Sequence Typing , Peptide Elongation Factor 1/genetics , Phenotype , RNA Polymerase II/genetics
5.
Mycopathologia ; 183(6): 859-877, 2018 Dec.
Article in English | MEDLINE | ID: mdl-30506286

ABSTRACT

Mycopathologia was founded in 1938 to 'diffuse the understanding of fungal diseases in man and animals among mycologists.' This was an important mission considering that pathogenic fungi for humans and animals represent a tiny minority of the estimated 1.5-5 million fungal inhabitants on Earth. These pathogens have diverged from the usual saprotrophic lifestyles of most fungi to colonize and infect humans and animals. Medical and veterinary mycology is the subdiscipline of microbiology that dwells into the mysteries of parasitic, fungal lifestyles. Among the oldest continuing scientific publications on the subject, Mycopathologia had its share of 'classic papers' since the first issue was published in 1938. An analysis of the eight decades of notable contributions reveals many facets of host-pathogen interactions among 183 volumes comprising about 6885 articles. We have analyzed the impact and relevance of this body of work using a combination of citation tools (Google Scholar and Scopus) since no single citation metric gives an inclusive perspective. Among the highly cited Mycopathologia publications, those on experimental mycology accounted for the major part of the articles (36%), followed by diagnostic mycology (16%), ecology and epidemiology (15%), clinical mycology (14%), taxonomy and classification (10%), and veterinary mycology (9%). The first classic publication, collecting nearly 200 citations, appeared in 1957, while two articles published in 2010 received nearly 150 citations each, which is notable for a journal covering a highly specialized field of study. An empirical analysis of the publication trends suggests continuing interests in novel diagnostics, fungal pathogenesis, review of clinical diseases especially with relevance to the laboratory scientists, taxonomy and classification of fungal pathogens, fungal infections and carriage in pets and wildlife, and changing ecology and epidemiology of fungal diseases around the globe. We anticipate that emerging and re-emerging fungal pathogens will continue to cause significant health burden in the coming decades. It remains vital that scientists and physicians continue to collaborate by learning each other's language for the study of fungal diseases, and Mycopathologia will strive to be their partner in this increasingly important endeavor to its 100th anniversary in 2038 and beyond.


Subject(s)
Bibliometrics , Fungi/physiology , Host-Pathogen Interactions , Mycology/history , Mycoses/microbiology , Mycoses/veterinary , Periodicals as Topic , Animals , History, 20th Century , History, 21st Century , Humans , Retrospective Studies
6.
BMC Genomics ; 18(1): 735, 2017 Sep 18.
Article in English | MEDLINE | ID: mdl-28923029

ABSTRACT

BACKGROUND: The Fusarium oxysporum species complex (FOSC) contains several phylogenetic lineages. Phylogenetic studies identified two to three major clades within the FOSC. The mitochondrial sequences are highly informative phylogenetic markers, but have been mostly neglected due to technical difficulties. RESULTS: A total of 61 complete mitogenomes of FOSC strains were de novo assembled and annotated. Length variations and intron patterns support the separation of three phylogenetic species. The variable region of the mitogenome that is typical for the genus Fusarium shows two new variants in the FOSC. The variant typical for Fusarium is found in members of all three clades, while variant 2 is found in clades 2 and 3 and variant 3 only in clade 2. The extended set of loci analyzed using a new implementation of the genealogical concordance species recognition method support the identification of three phylogenetic species within the FOSC. Comparative analysis of the mitogenomes in the FOSC revealed ongoing mitochondrial recombination within, but not between phylogenetic species. CONCLUSIONS: The recombination indicates the presence of a parasexual cycle in F. oxysporum. The obstacles hindering the usage of the mitogenomes are resolved by using next generation sequencing and selective genome assemblers, such as GRAbB. Complete mitogenome sequences offer a stable basis and reference point for phylogenetic and population genetic studies.


Subject(s)
Fusarium/genetics , Genome, Mitochondrial/genetics , Recombination, Genetic , Conserved Sequence , Genetic Variation , Genomics , Introns/genetics , Phylogeny
7.
PLoS Comput Biol ; 12(6): e1004753, 2016 06.
Article in English | MEDLINE | ID: mdl-27308864

ABSTRACT

GRAbB (Genomic Region Assembly by Baiting) is a new program that is dedicated to assemble specific genomic regions from NGS data. This approach is especially useful when dealing with multi copy regions, such as mitochondrial genome and the rDNA repeat region, parts of the genome that are often neglected or poorly assembled, although they contain interesting information from phylogenetic or epidemiologic perspectives, but also single copy regions can be assembled. The program is capable of targeting multiple regions within a single run. Furthermore, GRAbB can be used to extract specific loci from NGS data, based on homology, like sequences that are used for barcoding. To make the assembly specific, a known part of the region, such as the sequence of a PCR amplicon or a homologous sequence from a related species must be specified. By assembling only the region of interest, the assembly process is computationally much less demanding and may lead to assemblies of better quality. In this study the different applications and functionalities of the program are demonstrated such as: exhaustive assembly (rDNA region and mitochondrial genome), extracting homologous regions or genes (IGS, RPB1, RPB2 and TEF1a), as well as extracting multiple regions within a single run. The program is also compared with MITObim, which is meant for the exhaustive assembly of a single target based on a similar query sequence. GRAbB is shown to be more efficient than MITObim in terms of speed, memory and disk usage. The other functionalities (handling multiple targets simultaneously and extracting homologous regions) of the new program are not matched by other programs. The program is available with explanatory documentation at https://github.com/b-brankovics/grabb. GRAbB has been tested on Ubuntu (12.04 and 14.04), Fedora (23), CentOS (7.1.1503) and Mac OS X (10.7). Furthermore, GRAbB is available as a docker repository: brankovics/grabb (https://hub.docker.com/r/brankovics/grabb/).


Subject(s)
Genomics/methods , Software , Algorithms , Computational Biology , Computer Simulation , DNA, Fungal/genetics , DNA, Ribosomal/genetics , Fusarium/genetics , Genome, Fungal , Genome, Mitochondrial , Genomics/statistics & numerical data , High-Throughput Nucleotide Sequencing/statistics & numerical data
8.
Med Mycol ; 54(3): 287-94, 2016 Mar.
Article in English | MEDLINE | ID: mdl-26705832

ABSTRACT

Fusarium species are reported frequently as the most common causative agents of fungal keratitis in tropical countries such as India. Sixty-five fusaria isolated from patients were subjected to multilocus DNA sequencing to characterize the spectrum of the species associated with keratitis infections in India. Susceptibilities of these fusaria to ten antifungals were determined in vitro by the broth microdilution method. An impressive phylogenetic diversity of fusaria was reflected in susceptibilities differing at species level. Typing results revealed that the isolates were distributed among species in the species complexes (SCs) of F. solani (FSSC; n = 54), F. oxysporum (FOSC; n = 1), F. fujikuroi (FFSC; n = 3), and F. dimerum (FDSC; n = 7). Amphotericin B, voriconazole, and clotrimazole proved to be the most effective drugs, followed by econazole.


Subject(s)
Drug Resistance, Fungal , Eye Infections, Fungal/microbiology , Fusariosis/microbiology , Fusarium/classification , Fusarium/drug effects , Keratitis/microbiology , Phylogeny , Antifungal Agents/pharmacology , Fusarium/genetics , Fusarium/isolation & purification , Genetic Variation , Genotype , Humans , India , Microbial Sensitivity Tests , Multilocus Sequence Typing
9.
BMC Infect Dis ; 16: 148, 2016 Apr 14.
Article in English | MEDLINE | ID: mdl-27074951

ABSTRACT

BACKGROUND: Trichosporon species are ubiquitously spread and known to be part of the normal human flora of the skin and gastrointestinal tract. Trichosporon spp. normally cause superficial infections. However, in the past decade Trichosporon spp. are emerging as opportunistic agents of invasive fungal infections, particularly in severely immunocompromised patients. Clinical isolates are usually sensitive to triazoles, but strains resistant to multiple triazoles have been reported. CASE PRESENTATION: We report a high-level pan-azole resistant Trichosporon dermatis isolate causing an invasive cholangitis in a patient after liver re-transplantation. This infection occurred despite of fluconazole and low dose amphotericin B prophylaxis, and treatment with combined liposomal amphotericin B and voriconazole failed. CONCLUSION: This case and recent reports in literature show that not only bacteria are evolving towards pan-resistance, but also pathogenic yeasts. Prudent use of antifungals is important to withstand emerging antifungal resistance.


Subject(s)
Trichosporonosis/diagnosis , Amphotericin B/therapeutic use , Antifungal Agents/therapeutic use , Cholangitis/diagnosis , Cholangitis/drug therapy , Cholangitis/microbiology , Drug Resistance, Fungal , Hepatic Encephalopathy/diagnosis , Humans , Liver Cirrhosis/therapy , Liver Transplantation , Male , Microbial Sensitivity Tests , Middle Aged , Peritonitis/diagnosis , Phylogeny , Trichosporon/classification , Trichosporon/drug effects , Trichosporon/isolation & purification , Trichosporonosis/drug therapy , Trichosporonosis/microbiology , Voriconazole/therapeutic use
10.
Eukaryot Cell ; 14(2): 158-69, 2015 Feb.
Article in English | MEDLINE | ID: mdl-25480940

ABSTRACT

Sporotrichosis is one of the most frequent subcutaneous fungal infections in humans and animals caused by members of the plant-associated, dimorphic genus Sporothrix. Three of the four medically important Sporothrix species found in Brazil have been considered asexual as no sexual stage has ever been reported in Sporothrix schenckii, Sporothrix brasiliensis, or Sporothrix globosa. We have identified the mating type (MAT) loci in the S. schenckii (strain 1099-18/ATCC MYA-4821) and S. brasiliensis (strain 5110/ATCC MYA-4823) genomes by using comparative genomic approaches to determine the mating type ratio in these pathogen populations. Our analysis revealed the presence of a MAT1-1 locus in S. schenckii while a MAT1-2 locus was found in S. brasiliensis representing genomic synteny to other Sordariomycetes. Furthermore, the components of the mitogen-activated protein kinase (MAPK)-pheromone pathway, pheromone processing enzymes, and meiotic regulators have also been identified in the two pathogens, suggesting the potential for sexual reproduction. The ratio of MAT1-1 to MAT1-2 was not significantly different from 1:1 for all three Sporothrix species, but the population of S. brasiliensis in the outbreaks originated from a single mating type. We also explored the population genetic structure of these pathogens using sequence data of two loci to improve our knowledge of the pattern of geographic distribution, genetic variation, and virulence phenotypes. Population genetics data showed significant population differentiation and clonality with a low level of haplotype diversity in S. brasiliensis isolates from different regions of sporotrichosis outbreaks in Brazil. In contrast, S. schenckii isolates demonstrated a high degree of genetic variability without significant geographic differentiation, indicating the presence of recombination. This study demonstrated that two species causing the same disease have contrasting reproductive strategies and genetic variability patterns.


Subject(s)
Genes, Mating Type, Fungal/genetics , Reproduction, Asexual , Sporothrix/genetics , Sporotrichosis/transmission , Animals , Brazil , Cats , Disease Outbreaks , Humans , MAP Kinase Signaling System , Polymorphism, Genetic , Sporothrix/pathogenicity , Sporothrix/physiology , Sporotrichosis/veterinary , Virulence/genetics
11.
Mycoses ; 59(11): 705-709, 2016 Nov.
Article in English | MEDLINE | ID: mdl-27292696

ABSTRACT

The two most common filamentous fungi causing mycotic keratitis are Aspergillus and Fusarium spp. Around 70 Fusarium spp. are involved in causing human infections. In this study, four cases of keratitis in sugarcane farmers in India are being reported, caused by the sugar cane pathogen Fusarium sacchari, a species of the Fusarium fujikuroi species complex. Fusarial keratitis was established by potassium hydroxide/Calcofluor white wet mounts and fungal culture of corneal scrapings on conventional media. Final identification was done by genetic sequencing at CBS-KNAW, Utrecht, The Netherlands. The antifungal susceptibility testing was done using broth microdilution method as per CLSI document M38-A2. Four cases of F. sacchari keratitis were identified. Three of them had trauma with sugarcane leaves, whereas one sugarcane farmer reported trauma by vegetative matter. The morphological similarities among various Fusarium species warrant use of molecular methods for identification of cryptic species. A wide distribution of sugarcane farming could be the possible explanation for emergence of F. sacchari keratitis in India.


Subject(s)
Agricultural Workers' Diseases/microbiology , Eye Infections, Fungal/microbiology , Farmers , Fusarium/isolation & purification , Keratitis/microbiology , Saccharum/microbiology , Adult , Antifungal Agents/pharmacology , Cornea/microbiology , Female , Fusarium/drug effects , Fusarium/genetics , Fusarium/ultrastructure , Humans , India , Keratitis/diagnosis , Male , Microbial Sensitivity Tests , Middle Aged , Plant Leaves/microbiology , Sequence Analysis, DNA , Wounds and Injuries/complications , Wounds and Injuries/microbiology
12.
Mycopathologia ; 181(3-4): 161-3, 2016 Apr.
Article in English | MEDLINE | ID: mdl-26966007

ABSTRACT

Fusarium species are emerging human pathogens, next to being plant pathogens. Problems with Fusarium are in their diagnostics and in their difficult treatment, but also in what are actual Fusarium species or rather Fusarium-like species. In this issue Guevara-Suarez et al. (Mycopathologia. doi: 10.1007/s11046-016-9983-9 , 2016) characterized 89 isolates of Fusarium from Colombia showing especially lineages within the Fusarium solani and oxysporum species complexes to be responsible for onychomycosis.


Subject(s)
Fusarium/classification , Onychomycosis , Humans
13.
Mycopathologia ; 181(7-8): 497-504, 2016 Aug.
Article in English | MEDLINE | ID: mdl-27138574

ABSTRACT

Onychomycosis refers to fungal infection of the nail and is commonly caused by dermatophytes, while yeasts and non-dermatophytic molds (NDM) are increasingly recognized as pathogens in nail infections. The present study was done to delineate molecular epidemiology of Fusarium onychomycosis in India. Five hundred nail samples of Indian patients clinically suspected of onychomycosis were subjected to direct microscopy and fungal culture. Representative Fusarium isolates were further identified to species level by multi-locus sequencing for internal transcribed spacer, translation elongation factor 1 alpha (tef1-α) and RNA polymerase II subunit (rpb2) regions (primer pairs: ITS1/ITS4, EF1/EF2, 5f2/7cr, respectively). These representative strains were also tested for in vitro antifungal susceptibility by the broth microdilution method. Members of the genus Fusarium proved to be the most common NDM responsible for onychomycosis. The Fusarium spp. responsible for onychomycosis belonged to the Fusarium solani species complex (F. keratoplasticum and F. falciforme) and Fusarium fujikuroi species complex (F. proliferatum, F. acutatum and F. sacchari). Antifungal susceptibility results indicated that amphotericin B was the most effective antifungal across all isolates (MIC ranging 0.5-2 mg/L), followed by voriconazole (MIC ranging 1-8 µg/ml). However, a large variation was shown in susceptibility to posaconazole (MIC ranging 0.5 to >16 µg/ml). To conclude, we identified different Fusarium spp. responsible for onychomycosis in India with variation within species in susceptibility to antifungal agents, showing that fusariosis requires correct and prompt diagnosis as well as antifungal susceptibility testing.


Subject(s)
Antifungal Agents/pharmacology , Fusariosis/microbiology , Fusarium/classification , Fusarium/drug effects , Genotype , Onychomycosis/microbiology , Adult , Cluster Analysis , DNA, Fungal/chemistry , DNA, Fungal/genetics , DNA, Ribosomal Spacer/chemistry , DNA, Ribosomal Spacer/genetics , Female , Fusariosis/epidemiology , Fusarium/isolation & purification , Genotyping Techniques , Humans , India/epidemiology , Male , Microbial Sensitivity Tests , Middle Aged , Molecular Epidemiology , Multilocus Sequence Typing , Onychomycosis/epidemiology , Peptide Elongation Factor 1/genetics , Phylogeny , RNA Polymerase II/genetics , Young Adult
14.
Mol Biol Evol ; 31(4): 962-74, 2014 Apr.
Article in English | MEDLINE | ID: mdl-24448643

ABSTRACT

In filamentous fungi, allorecognition takes the form of heterokaryon incompatibility, a cell death reaction triggered when genetically distinct hyphae fuse. Heterokaryon incompatibility is controlled by specific loci termed het-loci. In this article, we analyzed the natural variation in one such fungal allorecognition determinant, the het-c heterokaryon incompatibility locus of the filamentous ascomycete Podospora anserina. The het-c locus determines an allogenic incompatibility reaction together with two unlinked loci termed het-d and het-e. Each het-c allele is incompatible with a specific subset of the het-d and het-e alleles. We analyzed variability at the het-c locus in a population of 110 individuals, and in additional isolates from various localities. We identified a total of 11 het-c alleles, which define 7 distinct incompatibility specificity classes in combination with the known het-d and het-e alleles. We found that the het-c allorecognition gene of P. anserina is under diversifying selection. We find a highly unequal allele distribution of het-c in the population, which contrasts with the more balanced distribution of functional groups of het-c based on their allorecognition function. One explanation for the observed het-c diversity in the population is its function in allorecognition. However, alleles that are most efficient in allorecognition are rare. An alternative and not exclusive explanation for the observed diversity is that het-c is involved in pathogen recognition. In Arabidopsis thaliana, a homolog of het-c is a pathogen effector target, supporting this hypothesis. We hypothesize that the het-c diversity in P. anserina results from both its functions in pathogen-defense, and allorecognition.


Subject(s)
Carrier Proteins/genetics , Fungal Proteins/genetics , Podospora/genetics , Amino Acid Sequence , Carrier Proteins/chemistry , Codon , Fungal Proteins/chemistry , Gene Frequency , Genetic Loci , Microbial Interactions , Molecular Sequence Data , Polymorphism, Genetic , Selection, Genetic
15.
J Antimicrob Chemother ; 70(4): 1068-71, 2015 Apr.
Article in English | MEDLINE | ID: mdl-25538167

ABSTRACT

OBJECTIVES: The aim of the present study was to evaluate and assess the in vitro activity of eight drugs, including the new azole isavuconazole, against 81 strains representing 13 species of the Fusarium fujikuroi species complex. METHODS: A total of 81 Fusarium spp. isolates, within the F. fujikuroi species complex, were identified by molecular methods and tested according to CLSI M38-A2. Eight antifungal compounds, including the new azole isavuconazole, were tested. Isolates were selected to represent the widest variety of geographical regions and to include clinical as well as environmental strains. RESULTS: Susceptibility profiles differed between and within species, with Fusarium verticillioides showing the lowest MICs and Fusarium nygamai the highest MICs. Amphotericin B was the most active drug, followed by voriconazole, posaconazole, isavuconazole and natamycin. The remaining antifungals (fluconazole, itraconazole and micafungin) showed poor activity with MIC/minimum effective concentration values of ≥ 32, ≥ 16 and >8 mg/L, respectively. CONCLUSIONS: Resistance patterns in the F. fujikuroi species complex are species specific and therefore identification down to species level is important for the choice of antifungal treatment.


Subject(s)
Antifungal Agents/pharmacology , Environmental Microbiology , Fusariosis/microbiology , Fusarium/drug effects , Fusarium/isolation & purification , Drug Resistance, Fungal , Fusarium/classification , Humans , Microbial Sensitivity Tests
16.
Mycoses ; 58(1): 48-57, 2015 Jan.
Article in English | MEDLINE | ID: mdl-25530264

ABSTRACT

Fusarium species are emerging causative agents of superficial, cutaneous and systemic human infections. In a study of the prevalence and genetic diversity of 464 fungal isolates from a dermatological ward in Thailand, 44 strains (9.5%) proved to belong to the genus Fusarium. Species identification was based on sequencing a portion of translation elongation factor 1-alpha (tef1-α), rDNA internal transcribed spacer and RNA-dependent polymerase subunit II (rpb2). Our results revealed that 37 isolates (84%) belonged to the Fusarium solani species complex (FSSC), one strain matched with Fusarium oxysporum (FOSC) complex 33, while six others belonged to the Fusarium incarnatum-equiseti species complex. Within the FSSC two predominant clusters represented Fusarium falciforme and recently described F. keratoplasticum. No gender differences in susceptibility to Fusarium were noted, but infections on the right side of the body prevailed. Eighty-nine per cent of the Fusarium isolates were involved in onychomycosis, while the remaining ones caused paronychia or severe tinea pedis. Comparing literature data, superficial infections by FSSC appear to be prevalent in Asia and Latin America, whereas FOSC is more common in Europe. The available data suggest that Fusarium is a common opportunistic human pathogens in tropical areas and has significant genetic variation worldwide.


Subject(s)
Dermatomycoses/microbiology , Fusariosis/epidemiology , Fusariosis/microbiology , Fusarium/isolation & purification , Dermatomycoses/diagnosis , Dermatomycoses/epidemiology , Female , Fusariosis/diagnosis , Fusarium/classification , Genotyping Techniques , Humans , Male , Multilocus Sequence Typing , Onychomycosis/diagnosis , Onychomycosis/epidemiology , Onychomycosis/microbiology , Peptide Elongation Factor 1/genetics , Phylogeny , Thailand/epidemiology , Tinea Pedis/diagnosis , Tinea Pedis/epidemiology , Tinea Pedis/microbiology
17.
Mycopathologia ; 179(1-2): 119-24, 2015 Feb.
Article in English | MEDLINE | ID: mdl-25234793

ABSTRACT

Members of the Fusarium solani species complex (FSSC) are causing the majority of the fusariosis in humans. Disseminated fusariosis has a high mortality and is predominantly observed in patients with leukemia. Here, we present the case of a fatal infection by a Fusarium strain with a degenerated phenotype, in a patient with acute lymphatic leukemia. Multiple nasal and skin biopsies as well as blood cultures yielded fungal growth, while in direct and histopathological examination of biopsy material septate hyphae were visible. Initial colonies were white with slimy masses with microconidia reminiscent of Fusarium/Acremonium, but with conidiospore production directly on the hyphae. Multi-locus sequence typing discerned a pionnotal-morphologically degenerated-colony of the recently recognized F. petroliphilum as etiological agent. The culture returned to a typical F. solani species complex morphology only after several weeks of growth in culture. Antifungal susceptibility tests indicate amphotericin B as best drug for this FSSC member rather than any of the azoles or echinocandins.


Subject(s)
Antifungal Agents/therapeutic use , Fusariosis/drug therapy , Fusariosis/mortality , Fusarium/drug effects , Amphotericin B/therapeutic use , Cefepime , Cephalosporins/therapeutic use , Clarithromycin/therapeutic use , Drug Resistance, Fungal , Female , Fusariosis/microbiology , Fusarium/classification , Humans , Levofloxacin/therapeutic use , Microbial Sensitivity Tests , Middle Aged , Multilocus Sequence Typing , Mycological Typing Techniques , Precursor Cell Lymphoblastic Leukemia-Lymphoma , Trimethoprim, Sulfamethoxazole Drug Combination/therapeutic use
18.
BMC Infect Dis ; 14: 588, 2014 Nov 12.
Article in English | MEDLINE | ID: mdl-25388601

ABSTRACT

BACKGROUND: Fusarium species are among the most common fungi present in the environment and some species have emerged as major opportunistic fungal infection in human. However, in immunocompromised hosts they can be virulent pathogens and can cause death. The pathogenesis of this infection relies on three factors: colonization, tissue damage, and immunosuppression. A novel Fusarium species is reported for the first time from keratitis in an agriculture worker who acquired the infection from plant material of maize. Maize plants are the natural host of this fungus where it causes stalk rot and seeding malformation under temperate and humid climatic conditions. The clinical manifestation, microbiological morphology, physiological features and molecular data are described. METHODS: Diagnosis was established by using polymerase chain reaction of fungal DNA followed by sequencing portions of translation elongation factor 1 alpha (TEF1 α) and beta-tubulin (BT2) genes. Susceptibility profiles of this fungus were evaluated using CLSI broth microdilution method. RESULTS: The analyses of these two genes sequences support a novel opportunist with the designation Fusarium temperatum. Phylogenetic analyses showed that the reported clinical isolate was nested within the Fusarium fujikuroi species complex. Antifungal susceptibility testing demonstrated that the fungus had low MICs of micafungin (0.031 µg/ml), posaconazole (0.25 µg/ml) and amphotericin B (0.5 µg/ml). CONCLUSION: The present case extends the significance of the genus Fusarium as agents of keratitis and underscores the utility of molecular verification of these emerging fungi in the human host.


Subject(s)
Antifungal Agents/therapeutic use , Fusarium/isolation & purification , Keratitis/diagnosis , Mycoses/microbiology , Zea mays/microbiology , Amphotericin B/pharmacology , Base Sequence , DNA, Fungal/chemistry , DNA, Fungal/genetics , Echinocandins/pharmacology , Fungal Proteins/genetics , Fusarium/classification , Fusarium/genetics , Humans , Keratitis/microbiology , Lipopeptides/pharmacology , Micafungin , Microbial Sensitivity Tests , Molecular Sequence Data , Multilocus Sequence Typing , Phylogeny , Sequence Analysis, DNA , Treatment Outcome , Triazoles/pharmacology , Tubulin/genetics
19.
Mycoses ; 57(4): 249-55, 2014 Apr.
Article in English | MEDLINE | ID: mdl-24612042

ABSTRACT

Disseminated infections caused by members of the Fusarium fujikuroi species complex (FFSC) occur regularly in immunocompromised patients. Here, we present the first human case caused by FFSC-member Fusarium andiyazi. Fever, respiratory symptoms and abnormal computerised tomography findings developed in a 65-year-old man with acute myelogenous leukaemia who was under posaconazole prophylaxis during his remission-induction chemotherapy. During the course of infection, two consecutive blood galactomannan values were found to be positive, and two blood cultures yielded strains resembling Fusarium species, according to morphological appearance. The aetiological agent proved to be F. andiyazi based on multilocus sequence typing. The sequencing of the internal transcribed spacer region did not resolve the closely related members of the FFSC, but additional data on partial sequence of transcription elongation factor 1 alpha subunit did. A detailed morphological study confirmed the identification of F. andiyazi, which had previously only been reported as a plant pathogen affecting various food crops.


Subject(s)
Antigens, Fungal/analysis , Aspergillus/chemistry , Fusariosis/diagnosis , Fusariosis/pathology , Fusarium/chemistry , Fusarium/isolation & purification , Mannans/analysis , Aged , Cross Reactions , DNA, Fungal/chemistry , DNA, Fungal/genetics , DNA, Ribosomal Spacer/chemistry , DNA, Ribosomal Spacer/genetics , Diagnosis, Differential , Fusariosis/microbiology , Fusarium/classification , Fusarium/genetics , Galactose/analogs & derivatives , Humans , Immunoenzyme Techniques/methods , Leukemia, Myeloid, Acute/complications , Leukemia, Myeloid, Acute/drug therapy , Male , Multilocus Sequence Typing , Tomography, X-Ray Computed
20.
Mycotoxin Res ; 2024 Apr 22.
Article in English | MEDLINE | ID: mdl-38647834

ABSTRACT

Aspergillus section Flavi (Flavi) is a diverse group of fungal species whose common members include A. flavus and A. parasiticus. These are well-known for the production of aflatoxin (AF) B and G and other toxic metabolites, like cyclopiazonic acid (CPA). They are saprophytic soil dwellers and also become crop opportunistic epiphytes. The consequence is contamination of the crop with mycotoxins, such as carcinogenic AF. We investigated the Flavi community structure of maize and that of their surrounding soil, including their mycotoxigenicity. Furthermore, we investigated the link of the maize Flavi diversity with preharvest maize AF levels. The study was carried out in four selected districts of Zambia, in a low rainfall zone. The Flavi characterisation was triphasic, involving morphological (colony colour and sclerotia formation), metabolic (AF and CPA production) and genetic (calmodulin gene polymorphism) analyses. Flavi abundance was determined by dilution plate technique on modified rose Bengal agar. Results showed that Flavi communities on maize and in soil differed. Maize had a higher Flavi species diversity than soil. A. parasiticus dominated the soil community by frequency of field appearance (85%), while maize was dominated by A. minisclerotigenes (45%). CPA-producers with or without AF production dominated the maize (65%) while producers of only AF (B/G) dominated the soil (88%). The ratio between maize A. parasiticus and A. minisclerotigenes abundance seemed to have had a bearing on the levels of AF in maize, with a ratio close to 1:1 having higher levels than a pure community of either A. parasiticus or A. minisclerotigenes.

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