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BACKGROUND: Fungal keratitis is a severe eye infection that can result in blindness and visual impairment, particularly in developing countries. Fusarium spp. are the primary causative agents of this condition. Diagnosis of Fusarium keratitis (FK) is challenging, and delayed treatment can lead to serious complications. However, there is limited epidemiological data on FK, especially in tropical areas. OBJECTIVES: This study aimed to describe the clinical, laboratorial and epidemiological characteristics of FK in a tropical semi-arid region of Brazil. PATIENTS/METHODS: Adult patients with laboratory-confirmed FK diagnosed between October 2019 and March 2022 were evaluated. Fusarium isolates were characterized at molecular level and evaluated regarding antifungal susceptibility. RESULTS: A total of 226 clinical samples from patients suspected of keratitis were evaluated; fungal growth was detected in 50 samples (22.12%); out of which 42 were suggestive of Fusarium spp. (84%). Molecular analysis of a randomly selected set of 27 isolates identified F. solani species complex (n = 14); F. fujikuroi sensu lato (n = 6) and F. dimerum sensu lato (n = 7); a total of 10 haplotypes were identified among the strains. All but one Fusarium strains were inhibited by amphotericin B, natamycin and fluconazole. Most patients were male (71.42%; 30 out of 42), aged from 27 to 73 years old. Trauma was the most important risk factor for FK (40.47%; 17 out of 42). Patients were treated with antifungals, corticoids and antibiotics; keratoplasty and eye enucleation were also performed. CONCLUSIONS: The study provided insights into the characteristics of FK in tropical regions and emphasized the importance of enhanced surveillance and management strategies.
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Antifúngicos , Infecciones Fúngicas del Ojo , Fusariosis , Fusarium , Queratitis , Pruebas de Sensibilidad Microbiana , Humanos , Brasil/epidemiología , Fusarium/genética , Fusarium/efectos de los fármacos , Fusarium/aislamiento & purificación , Fusarium/clasificación , Masculino , Femenino , Antifúngicos/farmacología , Antifúngicos/uso terapéutico , Adulto , Queratitis/microbiología , Queratitis/epidemiología , Queratitis/tratamiento farmacológico , Persona de Mediana Edad , Fusariosis/microbiología , Fusariosis/epidemiología , Fusariosis/tratamiento farmacológico , Infecciones Fúngicas del Ojo/microbiología , Infecciones Fúngicas del Ojo/epidemiología , Infecciones Fúngicas del Ojo/tratamiento farmacológico , Anciano , Adulto Joven , Adolescente , Clima Tropical , Anciano de 80 o más Años , Anfotericina B/farmacología , Anfotericina B/uso terapéuticoRESUMEN
Although Fusarium spp. rarely cause infections in healthy people, they can cause fusariosis, particularly in neutropenic hematological malignancies, bone marrow transplant patients, and immunocompromised patients, such as those with acquired immune deficiency syndrome (AIDS), and rarely in solid organ transplant recipients. Here, we present a case of a liver transplant recipient with F. solani species complex (FSSC) infection treated with posaconazole. A 61-year-old man presented with multiple itchy, painful, palpable, irregular, subcutaneous nodules on the right leg and total dystrophic onychomycosis in the right toenails. Incisional skin biopsies of the lesions were performed, and the samples were sent to the pathology and mycology laboratories for analysis. The clinical isolate was identified as FSSC using phenotypic, matrix-assisted laser desorption/ionization-time of flight mass spectrometry, and genotypic methods. Liposomal amphotericin B could not be administered owing to the development of side effects; hence, the patient was treated with posaconazole for 4 months. While some nodular lesions disappeared completely under this treatment, the others showed dimensional regression. This is the first case of FSSC infection with skin and nail involvement in a non-neutropenic, liver transplant patient in Turkey. Fusariosis may develop with rare species, such as FSSC, as first reported in this case of a liver transplant patient. Regardless of the species, amphotericin B is the first choice for treating fusariosis; however, posaconazole is an effective and safe alternative to amphotericin B.
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Fusariosis , Fusarium , Trasplante de Hígado , Masculino , Humanos , Persona de Mediana Edad , Fusariosis/diagnóstico , Fusariosis/tratamiento farmacológico , Anfotericina B/uso terapéutico , Antifúngicos/uso terapéuticoRESUMEN
Fungal-infections are mostly due to fungi in an adhering, biofilm-mode of growth and not due to planktonically growing, suspended-fungi. 1, 8-cineole is a natural product, which has been shown to possess antifungal effect. However, the anti-biofilm effect and mechanism of 1,8-cineole against Fusarium solani species complex has not reported previously. In this study, we found that 1,8-cineole has a good antifungal activity against F. solani with an MIC value of 46.1 µg/ml. Notably, 1,8-cineole showed good anti-biofilm formation activity against F. solani via inhibiting cell adhesion, hypha formation and decreasing the secretion of extracellular matrix at the concentration of ≥5.76 µg/ml. In addition, transcriptome sequencing analysis results showed that F. solani species complex genes related to ECM, protein synthesis and energy metabolism were down-expressed in the biofilms formation process treated with 1,8-cineole. In conclusion, these results show that 1,8-cineole has good anti-biofilm formation activity against F. solani species complex, and it exerts its anti-biofilm formation activity by downregulating of ergosterol biosynthetic genes, inhibiting adhesion, hindering the synthesis of ECM and interfering mitochondrial activity. This study suggests that 1,8-cineole is a promising anti-biofilm agent against F. solani species complex.
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Fusarium is a phytopathogenic fungus involved in human pathology and is present in space stations. It is essential to understand the effects of microgravity on the physiology of this fungus to determine the potential risks to the health of crew members and to propose the necessary countermeasures. This study aimed to determine changes in the physiological parameters of the Fusarium solani species complex under simulated microgravity generated using a random positioning machine (RPM) and phenotypic approaches. We observed increased growth, spore production, and germination while biofilm production was reduced under RPM exposure. These in vitro data show the importance of further studying this fungus as it has been repeatedly demonstrated that microgravity weakens the immune system of astronauts.
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BACKGROUND: The Fusarium solani species complex (FSSC) comprises fungal pathogens responsible for mortality in a diverse range of animals and plants, but their genome diversity and transcriptome responses in animal pathogenicity remain to be elucidated. We sequenced, assembled and annotated six chromosome-level FSSC clade 3 genomes of aquatic animal and plant host origins. We established a pathosystem and investigated the expression data of F. falciforme and F. keratoplasticum in Chinese softshell turtle (Pelodiscus sinensis) host. RESULTS: Comparative analyses between the FSSC genomes revealed a spectrum of conservation patterns in chromosomes categorised into three compartments: core, fast-core (FC), and lineage-specific (LS). LS chromosomes contribute to variations in genomes size, with up to 42.2% of variations between F. vanettenii strains. Each chromosome compartment varied in structural architectures, with FC and LS chromosomes contain higher proportions of repetitive elements with genes enriched in functions related to pathogenicity and niche expansion. We identified differences in both selection in the coding sequences and DNA methylation levels between genome features and chromosome compartments which suggest a multi-speed evolution that can be traced back to the last common ancestor of Fusarium. We further demonstrated that F. falciforme and F. keratoplasticum are opportunistic pathogens by inoculating P. sinensis eggs and identified differentially expressed genes also associated with plant pathogenicity. These included the most upregulated genes encoding the CFEM (Common in Fungal Extracellular Membrane) domain. CONCLUSIONS: The high-quality genome assemblies provided new insights into the evolution of FSSC chromosomes, which also serve as a resource for studies of fungal genome evolution and pathogenesis. This study also establishes an animal model for fungal pathogens of trans-kingdom hosts.
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Fusarium , Animales , Fusarium/genética , Transcriptoma , Enfermedades de las Plantas/genética , Enfermedades de las Plantas/microbiología , Filogenia , Genómica , Plantas/genéticaRESUMEN
Immature stages of insects are vulnerable to various antagonists, including pathogens. While the abiotic factors affecting pathogen prevalence in insect populations are reasonably well documented, much less is known about relevant ecological interactions. We studied the probability of the larvae of three lepidopteran species to die from fungal infection as a function of insect species and food plants in central Argentina. Local free-growing food plants were used to feed the lepidopteran larvae. The prevalence of entomopathogenic fungi remained low (about 5%), which is a value well consistent with observations on similar systems in other regions. Eight fungal species recorded, primarily belonging to Fusarium and Aspergillus, add evidence to the reconsideration of the nutritional modes in these genera in distinguishing the role of some species (complexes) to cause insect infections. Food plant species were found to have a substantial effect on the prevalence of entomopathogenic fungi. This was especially clear for the most abundant fungal species, a representative of the Fusarium fujikuroi complex. Feeding on a particular plant taxon can thus have a specific fitness cost. Compared to the data collected from Northern Europe, the Argentinian assemblages from the families Aspergillaceae and Nectriaceae overlapped at the genus level but did not share species. It remains to be confirmed if this level of divergence in the composition of assemblages of entomopathogenic fungi among distant regions represents a global pattern.
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Onychomycosis, a nail fungal infection, is normally caused by dermatophytes. However, yeasts and non-dermatophyte moulds (NDM) are among pathogens that cause nail disease. Regarding, this study aimed to describe the molecular epidemiology of Fusarium onychomycosis in the North of Iran. Two hundred and fifty seven nail samples collected from the patients clinically suspected of onychomycosis were subjected to direct microscopy, calcofluor white staining and culture. Fusarium isolates were identified at a species level through determination of multi-locus sequences for internal transcribed spacer and translation elongation factor 1 alpha. Based on the findings, Fusarium species were isolated from onychomycosis patients (n = 27). According to a previous partial genes analysis, the species in the recent study belonged to the members of F. fujikuroi species complex (n = 14), Fusarium incarnatum-equiseti species complex (n = 1) and F. solani species complex (n = 12). In this study, F. proliferatum was the dominant Fusarium species collected from the samples. The correct identification of Fusarium species is essential regarding the increased prevalence of Fusarium onychomycosis and the inherent resistance of these agents to a wide spectrum of antifungals. The obtained results indicated variation in the epidemiology of Fusarium species isolated from onychomycosis. Moreover, the minimum inhibitory concentration (MIC) of luliconazole and lanoconazole was in the range of 0.001-1 µg/ml, with the geometric mean of MICs obtained at 0.0103 and 0.0343 µg/ml against Fusarium species, respectively. These findings can increase researchers' knowledge regarding diversity of species, distribution of onychomycosis and the choice of a proper treatment.
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Fusarium , Onicomicosis , Antifúngicos/farmacología , Variación Genética , Humanos , Irán/epidemiología , Onicomicosis/epidemiología , Onicomicosis/microbiología , Factor 1 de Elongación Peptídica/genética , PrevalenciaRESUMEN
Sweetpotato (Ipomoea batatas) is the eighth most important crop globally. However, the production and quality of sweetpotatoes are threatened by Fusarium diseases that are prevalent around the world. In this study, a Fusarium species that causes root and stem rot in sweetpotatoes was studied. The pathogenic fungus CRI 24-3 was isolated and sequenced using third- and next-generation sequencing techniques and a 49.6 Mb chromosome-level draft genome containing 15,374 putative coding genes were obtained. Molecular phylogenetic analysis showed that CRI 24-3 was an F. solani-melongenae strain within clade 3 of the F. solani species complex (FSSC). CRI 24-3 showed a relatively high number of virulence factors, such as carbohydrate-active enzymes (CAZymes), pathogen-host interaction (PHI) proteins, and terpene synthases (TSs), compared with the number of those identified in other sequenced FSSC members. Comparative genome analysis revealed considerable conservation and unique characteristics between CRI 24-3 and other FSSC species. In conclusion, the findings in the current study provide important genetic information about F. solani-melongenae and should be useful in the exploration of pathogenicity mechanisms and the development of Fusarium disease management strategies. IMPORTANCE Fusarium root and stem rot in sweetpotato are prevalent in the main sweetpotato-growing areas in China, and fungal isolation, morphological characteristics, and molecular phylogenetic analysis of the disease causal agent (F. solani-melongenae isolate CRI 24-3) were systematically studied. The genome sequence of F. solani-melongenae isolates CRI 24-3 was first reported, which should provide a basis for genome assembly of other closely related Fusarium species. Carbohydrate-active enzymes predicted in CRI 24-3 may be important to convert the substantial polysaccharides to sustainable and renewable energy. Moreover, other virulence factors facilitating Fusarium diseases, including effectors and toxic secondary metabolites, are ideal objects for pathogenicity mechanism research and molecular targets for fungicide development. The findings of comparative genome analysis of CRI 24-3 and 15 sequenced members of the F. solani species complex help promote an integral understanding of genomic features and evolutionary relationships in Fusarium.
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Fusarium , Carbohidratos , Fusarium/genética , Filogenia , Enfermedades de las Plantas/microbiología , Factores de Virulencia/genéticaRESUMEN
A novel dsRNA mycovirus was found in Fusarium solani (F. solani) strain NW-FVA 2572. The fungus was originally isolated from a root, associated with stem collar necrosis of Fraxinus excelsior L. The viral genome is composed of four segments, which range from around 3.5 kbp to 1.7 kbp (RNA 1: 3522 bp; RNA 2: 2633 bp; RNA 3: 2403 bp; RNA 4: 1721 bp). The segments share a conserved and capped 5'-terminus and their 3'-termini are polyadenylated. Protein sequencing showed that the viral RdRP is encoded on segment 1. The virus clusters together with Aspergillus mycovirus 341 (AsV341), Aspergillus heteromorphus alternavirus 1 (AheAV1), Aspergillus foetidus virus-fast (AfV-F) and Cordyceps chanhua alternavirus 1 (CcAV1). As highest value, the RdRP showed 61.50% identical amino acids with P1 of the AfV-F. The capsid protein is encoded on segment 3, the proteins encoded on RNA 2 and RNA 4 are of unknown function. Segment 4 harbors large UTRs (186 nts at the 5'-terminus and 311 nts at the 3'-terminus). Based on its genome organization and phylogenetic position, the virus is suggested to be a new member of the proposed family Alternaviridae and was therefore named Fusarium solani alternavirus 1 (FsAV1). This is the first report of an Alternavirus infecting a fungus of the F. solani species complex (FSSC).
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Virus Fúngicos , Fusarium , Virus ARN , Virus no Clasificados , Fusarium/genética , Genoma Viral , Sistemas de Lectura Abierta , Filogenia , ARN Bicatenario/genética , ARN Viral/genética , ARN Polimerasa Dependiente del ARN , Virus no Clasificados/genéticaRESUMEN
The fungal pathogens Fusarium falciforme and Fusarium keratoplasticum are responsible for the sea turtle egg fusariosis (STEF) throughout main nesting areas of the world. In this study, we investigated whether eggs of the invasive alien red-eared slider turtle, Trachemys scripta, can carry these fungal pathogens. Using multilocus sequence typing of four nuclear DNA regions, we found that eggs of T. scripta naturally can carry these two Fusarium pathogenic species, as well as other Fusarium species belonging to the Fusarium solani species complex. Physiological studies on F. falciforme and F. keratoplasticum isolates revealed that their optimal growth temperature coincided with the pivotal temperature for T. scripta embryos, ca 29.5 ± 0.5 °C, providing an evidence of a potential advantageous biological property for host colonization and virulence. A host-pathogen interaction network analysis of species of the FSSC and their hosts confirmed that F. falciforme and F. keratoplasticum are generalist pathogens in a wide range of animal hosts of worldwide geographical distribution. Finally, we show that nesting areas of this invasive turtle T. scripta in the Mediterranean freshwater marshes can act as chronic reservoirs of these STEF pathogens, and this invasive species can act as a potential vector for the spread of STEF among wild native species and even to humans.
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Fusariosis , Tortugas , Animales , Agua Dulce , Fusariosis/microbiología , Especies Introducidas , Tipificación de Secuencias Multilocus , Tortugas/genética , Tortugas/microbiologíaRESUMEN
BACKGROUND: Succinate dehydrogenase inhibitors (SDHIs) have been widely used to manage plant diseases caused by phytopathogenic fungi. Although attention to and use of SDHI fungicides has recently increased, molecular responses of fungal pathogens to SDHIs have often not been investigated. A SDHI fungicide, fluopyram, has been used as a soybean seed treatment and has displayed effective control of Fusarium virguliforme, one of the causal agents of soybean sudden death syndrome. To examine genome-wide gene expression of F. virguliforme to fluopyram, RNA-seq analysis was conducted on two field strains of F. virguliforme with differing SDHI fungicide sensitivity in the absence and presence of fluopyram. RESULTS: The analysis indicated that several xenobiotic detoxification-related genes, such as those of deoxygenase, transferases and transporters, were highly induced by fluopyram. Among the genes, four ATP-binding cassette (ABC) transporters were characterized by the yeast expression system. The results revealed that expression of three ABCG transporters was associated with reduced sensitivity to multiple fungicides including fluopyram. In addition, heterologous expression of a major facilitator superfamily (MFS) transporter that was highly expressed in the fluopyram-insensitive F. virguliforme strain in the yeast system conferred decreased sensitivity to fluopyram. CONCLUSION: This study demonstrated that xenobiotic detoxification-related genes were highly upregulated in response to fluopyram, and expression of ABC or MFS transporter genes was associated with reduced sensitivity to the SDHI fungicide. This is the first transcriptomic analysis of the fungal species response to fluopyram and the finding will help elucidate the molecular mechanisms of SDHI resistance. © 2021 Society of Chemical Industry.
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Fungicidas Industriales , Fusarium , Enfermedades de las Plantas , Benzamidas/farmacología , Fungicidas Industriales/farmacología , Fusarium/genética , Fusarium/patogenicidad , Enfermedades de las Plantas/microbiología , Piridinas/farmacología , Glycine max/microbiología , Succinato Deshidrogenasa/antagonistas & inhibidores , Ácido SuccínicoRESUMEN
Background and Objectives: Fusarium species are known to be one of the common causes of keratitis. This study was conducted to identify Fusarium spp. causing keratitis and to investigate their genetic diversity using TEF1 and RPB2 gene sequences. Materials and Methods: Twenty-four clinical isolates of Fusarium were isolated from the patient with keratitis. Phylogenetic analysis of two-locus of the 24 clinical isolates and three reference strains was carried out using the maximum parsimony and RAxML methods. Results: Based on gene sequences of the 24 clinical isolates, 17, 4, and 3 isolates were identified as Fusarium solani species complex (FSSC), Fusarium fujikuroi species complex (FFSC), and Fusarium oxysporum, respectively. FFSC include F. proliferatum (n=1), F. globosum (n=1), F. verticillioides (n=1), and F. brevicatenulatum (n=1), respectively. Conclusion: Given that sequence of a sole gene can be challenging and on the other hand, due to the high resistance to antifungal drugs, identification of Fusarium species is of substantial significance. In this study, by designing a novel set of primers for the RPB2 area and using TEF1 primer, we were able to differentiate 24 Fusarium spp. isolated from patients with keratitis.
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Ficus carica plantations in Japan were first reported to be infested by an ambrosia beetle species, identified as Euwallacea interjectus, in 1996. The purpose of this study was to determine the symbiotic fungi of female adults of E. interjectus emerging from F. carica trees infected with fig wilt disease (FWD). Dispersal adults (51 females) of E. interjectus, which were collected from logs of an infested fig tree in Hiroshima Prefecture, Western Japan, were separated into three respective body parts (head, thorax, and abdomen) and used for fungal isolation. Isolated fungi were identified based on the morphological characteristics and DNA sequence data. Over 13 species of associated fungi were detected, of which a specific fungus, Fusarium kuroshium, was dominant in female head (including oral mycangia). The plant-pathogenic fungus of FWD, Ceratocystis ficicola, was not observed within any body parts of E. interjectus. We further discussed the relationship among E. interjectus and its associated fungi in fig tree.
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Chickpea (Cicer aretinium L.) is a legume crop of great importance worldwide. In January 2019, wilting symptoms on chickpea (stunted grow, withered leaves, root rot and wilted plants) were observed in three fields of Culiacan Sinaloa Mexico, with an incidence of 3 to 5%. To identify the cause, eighty symptomatic chickpea plants were sampled. Tissue from roots was plated on potato dextrose agar (PDA) medium. Typical Fusarium spp. colonies were obtained from all root samples. Ten pure cultures were obtained by single-spore culturing (Ff01 to Ff10). On PDA the colonies were abundant with white aerial mycelium, hyphae were branched and septae and light purple pigmentation was observed in the center of old cultures (Leslie and Summerell 2006). From 10-day-old cultures grown on carnation leaf agar medium, macroconidias were falciform, hyaline, with slightly curved apexes, three to five septate, with well-developed foot cells and blunt apical cells, and measured 26.6 to 45.8 × 2.2 to 7.0 µm (n = 40). The microconidia (n = 40) were hyaline, one to two celled, produced in false heads that measured 7.4 to 20.1 (average 13.7) µm × 2.4 to 8.9 (average 5.3) µm (n = 40) at the tips of long monophialides, and were oval or reniform, with apexes rounded, 8.3 to 12.1 × 1.6 to 4.7 µm; chlamydospores were not evident. These characteristics fit those of the Fusarium solani (Mart.) Sacc. species complex, FSSC (Summerell et al. 2003). The internal transcribed spacer and the translation elongation factor 1 alpha (EF1-α) genes (O'Donnell et al. 1998) were amplified by polymerase chain reaction and sequenced from the isolate Ff02 and Ff08 (GenBank accession nos. KJ501093 and MN082369). Maximum likelihood analysis was carried out using the EF1-α sequences (KJ501093 and MN082369) from the Ff02 and Ff08 isolates and other species from the Fusarium solani species complex (FSSC). Phylogenetic analysis revealed the isolate most closely related with F. falciforme (100% bootstrap). For pathogenicity testing, a conidial suspension (1x106 conidia/ml) was prepared by harvesting spores from 10-days-old cultures on PDA. Twenty 2-week-old chickpea seedlings from two cultivars (P-2245 and WR-315) were inoculated by dipping roots into the conidial suspension for 20 min. The inoculated plants were transplanted into a 50-hole plastic tray containing sterilized soil and maintained in a growth chamber at 25°C, with a relative humidity of >80% and a 12-h/12-h light/dark cycle. After 8 days, the first root rot symptoms were observed on inoculating seedlings and the infected plants eventually died within 3 to 4 weeks after inoculation. No symptoms were observed plants inoculated with sterilized distilled water. The fungus was reisolated from symptomatic tissues of inoculated plants and was identified by sequencing the partial EF1-α gene again and was identified as F. falciforme (FSSC 3 + 4) (O'Donnell et al. 2008) based on its morphological characteristics, genetic analysis, and pathogenicity test, fulfilling Koch's postulates. The molecular identification was confirmed via BLAST on the FusariumID and Fusarium MLST databases. Although FSSC has been previously reported causing root rot in chickpea in USA, Chile, Spain, Cuba, Iran, Poland, Israel, Pakistan and Brazil, to our knowledge this is the first report of root rot in chickpea caused by F. falciforme in Mexico. This is important for chickpea producers and chickpea breeding programs.
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BACKGROUND: Invasive fusariosis is associated with marked morbidity and mortality in immunocompromised hosts, and clinical outcomes are poor with conventional therapy. Olorofim (F901318) is an investigational antifungal in the orotomide class that selectively targets fungal dihydroorotate dehydrogenase (DHODH) causing inhibition of pyrimidine biosynthesis. OBJECTIVE: We evaluated the in vitro activity of olorofim against 61 clinical isolates of the Fusarium oxysporum and F solani species complexes (FOSC and FSSC, respectively), the most prevalent causes of invasive fusariosis. METHODS: Clinical isolates of FOSC (n = 45) and FSSC (n = 16) were identified using DNA sequence analysis of the translation elongation factor 1-alpha (TEF1α) and RNA polymerase II second largest subunit (RPB2). Antifungal susceptibility testing was performed by CLSI M38 broth microdilution for olorofim, amphotericin B, isavuconazole, posaconazole, voriconazole and micafungin. RESULTS: Olorofim demonstrated good in vitro activity against both FOSC and FSSC. Against the 45 FOSC isolates, olorofim MICs ranged between 0.03-0.5 mg/L and 0.06->4 mg/L at the 50% and 100% inhibition endpoints, respectively. Against FSSC isolates, olorofim MIC ranged between 0.25-1 mg/L and 1->4 mg/L at 50% and 100% inhibition, respectively. While amphotericin B also demonstrated similar in vitro activity (MIC ranges 1-4 and 0.25-4 mg/L against FOSC and FSSC, respectively), neither the triazoles nor micafungin demonstrated consistent in vitro activity against Fusarium isolates at clinically relevant concentrations. CONCLUSIONS: The investigational agent olorofim demonstrated good in vitro activity against FOSC and FSSC clinical isolates. Further studies are warranted to determine how well this in vitro activity translates into in vivo efficacy.
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Acetamidas/farmacología , Fusarium , Piperazinas/farmacología , Pirimidinas/farmacología , Pirroles/farmacología , Antifúngicos/farmacología , Fusariosis/tratamiento farmacológico , Fusariosis/microbiología , Fusarium/efectos de los fármacos , Fusarium/aislamiento & purificación , Humanos , Pruebas de Sensibilidad MicrobianaRESUMEN
BACKGROUND: The transition to a biobased economy involving the depolymerization and fermentation of renewable agro-industrial sources is a challenge that can only be met by achieving the efficient hydrolysis of biomass to monosaccharides. In nature, lignocellulosic biomass is mainly decomposed by fungi. We recently identified six efficient cellulose degraders by screening fungi from Vietnam. RESULTS: We characterized a high-performance cellulase-producing strain, with an activity of 0.06 U/mg, which was identified as a member of the Fusarium solani species complex linkage 6 (Fusarium metavorans), isolated from mangrove wood (FW16.1, deposited as DSM105788). The genome, representing nine potential chromosomes, was sequenced using PacBio and Illumina technology. In-depth secretome analysis using six different synthetic and artificial cellulose substrates and two agro-industrial waste products identified 500 proteins, including 135 enzymes assigned to five different carbohydrate-active enzyme (CAZyme) classes. The F. metavorans enzyme cocktail was tested for saccharification activity on pre-treated sugarcane bagasse, as well as untreated sugarcane bagasse and maize leaves, where it was complemented with the commercial enzyme mixture Accellerase 1500. In the untreated sugarcane bagasse and maize leaves, initial cell wall degradation was observed in the presence of at least 196 µg/mL of the in-house cocktail. Increasing the dose to 336 µg/mL facilitated the saccharification of untreated sugarcane biomass, but had no further effect on the pre-treated biomass. CONCLUSION: Our results show that F. metavorans DSM105788 is a promising alternative pre-treatment for the degradation of agro-industrial lignocellulosic materials. The enzyme cocktail promotes the debranching of biopolymers surrounding the cellulose fibers and releases reduced sugars without process disadvantages or loss of carbohydrates.
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Erythrina spp. trees have been declining since the 2000s worldwide, and fungi belonging to Fusarium solani species complex (FSSC) have been suggested to be a causal factor of decline and mortality of Erythrina variegata trees in Okinawa Island, Japan. In addition to the FSSC isolate grouped as "it-1" based on ITS sequence data (previously called strain A), we conducted an inoculation experiment with two isolates grouped as "it-2" (previously strain B), which is genetically close to it-1. Two it-2 isolates originating from two islands showed pathogenicity to E. variegata with the same symptoms as those caused by it-1 isolate. We also found the isolates of it-1 and it-2 were widely distributed, including on Ishigaki Island, â¼400 km south of Okinawa Island across the ocean. All isolates of it-1 and it-2 belong to the ambrosia Fusarium clade of the FSSC, a group of symbionts of ambrosia beetles, including the pathogens of Fusarium dieback in avocados and teas. The detection of ambrosia beetles Euwallacea spp. from our specimens provided information on the vectors of the pathogens. Our present results suggest the fungi of the FSSC could be responsible for the Erythrina decline in other areas with damage.
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Erythrina , Fusarium , Animales , Japón , Filogenia , Enfermedades de las Plantas , VirulenciaRESUMEN
Fungal keratitis (FK) is a site-threatening infection of the cornea associated with ocular trauma and contact lens wear. Members of the Fusarium solani species complex (FSSC) are predominant agents of FK worldwide, but genes that support their corneal virulence are poorly understood. As a means to bolster genetic analysis in FSSC pathogens, we sought to employ a CRISPR/Cas9 system in an FK isolate identified as Fusarium petroliphilum. Briefly, this approach involves the introduction of two components into fungal protoplasts: (1) A purified Cas9 protein complexed with guide RNAs that will direct the ribonuclease to cut on either side of the gene of interest, and (2) a "repair template" comprised of a hygromycin resistance cassette flanked by 40 bp of homology outside of the Cas9 cuts. In this way, Cas9-induced double strand breaks should potentiate double homologous replacement of the repair template at the desired locus. We targeted a putative ura3 ortholog since its deletion would result in an easily discernable uracil auxotrophy. Indeed, 10% of hygromycin-resistant transformants displayed the auxotrophic phenotype, all of which harbored the expected ura3 gene deletion. By contrast, none of the transformants from the repair template control (i.e., no Cas9) displayed the auxotrophic phenotype, indicating that Cas9 cutting was indeed required to promote homologous integration. Taken together, these data demonstrate that the in vitro Cas9 system is an easy and efficient approach for reverse genetics in FSSC organisms, including clinical isolates, which should enhance virulence research in these important but understudied ocular pathogens.
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Introduction. The remarkable intrinsic resistance of Fusarium species to most antifungal agents results in high mortality rates in the immunocompromised population.Aims. This study aimed to investigate the epidemiology, clinical features and antifungal susceptibility of Fusarium isolates in patients with invasive fusariosis.Methodology. A total of 27 patients admitted to a referral hospital from January 2008 to June 2017 were evaluated. Antifungal susceptibility testing of isolates was performed by broth microdilution according to the Clinical and Laboratory Standards Institute guidelines.Results. Haematological malignancy was the predominant underlying condition, with an incidence of invasive fusariosis of 14.8 cases per 1000 patients with acute lymphoid leukaemia and 13.1 cases per 1000 patients with acute myeloid leukaemia. The Fusarium solani species complex (FSSC) was the most frequent agent group, followed by the Fusarium oxysporum species complex (FOSC). Voriconazole showed the best activity against Fusarium, followed by amphotericin B. Itraconazole showed high minimum inhibitory concentration values, indicating in vitro resistance. Clinical FSSC isolates were significantly (P<0.05) more resistant to amphotericin B and voriconazole than FOSC isolates.Conclusion. The present antifungal susceptibility profiles indicate a high incidence of fusariosis in patients with haematological malignancy. Species- and strain-specific differences in antifungal susceptibility exist within Fusarium in this setting.