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Background: Fusarium keratitis accounts for around 50% of mycotic keratitis cases. Major virulence factors produced by keratopathogenic fungi are proteases. Objective: The aim of the current study was to identify proteases contributing to corneal pathogenicity of Fusarium species. Methods: Culture filtrates from fourteen Fusarium solani species complex (FSSC) isolates and three F. delphinoides isolates were evaluated for protease activity and gelatine zymography. Mass spectroscopy was carried out using a partially purified enzyme and total extracellular extract. Protease gene expression in an in-vitro condition and an ex-vivo goat corneal infection model was measured using qRT-PCR. Specific activity was observed in a wide range and at a broad pH range; and isolates Cs1 (maximum) and Cc50 (minimum) were selected for the infection model. Results: Gene expression in in-vitro condition showed the highest fold change for proteases (C7YY94, C7Z7U2 and C7Z6W1) while in an ex-vivo infection highest fold change was seen for proteases (C7Z6W1, C7YQJ2 and C7Z7U2); in decreasing order, respectively. Expression of aminopeptidase (C7Z6W1) was 50-fold higher in the infected cornea in both isolates (Cs1 and Cc50); while expression of carboxypeptidase (C7YVF3) was 15-fold higher only in isolate Cs1. Corneal histology showed less penetration of Cc50 than Cs1 into the stroma. Mass spectrometry showed the presence of carboxypeptidase (C7YVF3) and tripeptidyl amino peptidase. Conclusion: It can be concluded that clinical isolates of FSSC produce varying amounts of proteases and differ in specific activity and gene expression in both conditions (in vitro and ex vivo). Carboxypeptidase and aminopeptidase contribute to the pathogenic potential of Fusarium solani species complex.

Aim: To evaluate the inhibition of efflux pumps by using promethazine (PMZ) as a strategy to control Fusarium solani species complex (FSSC). Materials & methods: The susceptibility of FSSC strains to PMZ and the interaction between PMZ and antifungals were evaluated. The efflux pump activity was confirmed by flow cytometry with rhodamine 6G. Finally, PMZ was tested against FSSC biofilms. Results: PMZ inhibited FSSC planktonic growth and showed synergism with antifungals. PMZ reduced R6G efflux and inhibited cell adhesion, impaired the development of biofilms and disrupted mature biofilms. PMZ-challenged biofilms showed increased sensitivity to amphotericin B. Conclusion: The study provides indirect evidence of the occurrence of efflux pumps in FSSC and opens a perspective for this target in the control of fusariosis.

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.

This article is to alert medical mycologists and infectious disease specialists of recent name changes of medically important species of the filamentous mold Fusarium. Fusarium species can cause localized and life-threating infections in humans. Of the 70 Fusarium species that have been reported to cause infections, close to one-third are members of the Fusarium solani species complex (FSSC), and they collectively account for approximately two-thirds of all reported Fusarium infections., This article is to alert medical mycologists and infectious disease specialists of recent name changes of medically important species of the filamentous mold Fusarium. Fusarium species can cause localized and life-threating infections in humans. Of the 70 Fusarium species that have been reported to cause infections, close to one-third are members of the Fusarium solani species complex (FSSC), and they collectively account for approximately two-thirds of all reported Fusarium infections. Many of these species were recently given scientific names for the first time by a research group in the Netherlands, but they were misplaced in the genus Neocosmospora. In this paper, we present genetic arguments that strongly support inclusion of the FSSC in Fusarium. There are potentially serious consequences associated with using the name Neocosmospora for Fusarium species because clinicians need to be aware that fusaria are broadly resistant to the spectrum of antifungals that are currently available.

The genus Fusarium is largely represented in fungal infections, not only mostly in plants but also in humans. Fungi belonging to the Fusarium solani species complex (FSSC) are those that are most frequently isolated in invasive fusariosis and present elevated minimum inhibitory concentrations for the main antifungal drugs used in medicine. This study is the first to investigate the resistance mechanism in FSSC by monitoring CYP51A expression in the presence of azole antifungals. After exposure to voriconazole, an overinduction of CYP51A was observed irrespective of the concentration of antifungal used and the generation studied. The same observation is made for other azole antifungals, posaconazole and tebuconazole, but not for amphotericin B. This observation could contribute to explaining why some antifungal molecules used in agriculture or medical practices may have low susceptibility for some fungi.

Fusarium species are among the most commonly isolated causes of fungal keratitis. Most species of the genus Fusarium belong to Fusarium solani species complex (FSSC). Fusarium lichenicola, a member of the FSSC complex, is a well-established plant and human pathogen. However, reports of fungal keratitis due to Fusarium lichenicola have not been frequently reported. To the best of our knowledge, only twelve cases of Fusarium lichenicola keratitis have been reported in the past fifty years. Clinical cases of Fusarium lichenicola may have most likely been misidentified because of the lack of clinical and microbiological suspicion, as well as inadequate diagnostic facilities in many tropical countries where the burden of the disease may be the highest. We report a case of fungal keratitis caused by Fusarium lichenicola and present a global review of the literature of all cases of fungal keratitis caused by this potentially blinding fungus.

In Japan, Fusarium species are known etiological agents of human fungal infection; however, there has been no report of a large-scale epidemiological study on the etiological agents of fusariosis. A total of 73 Fusarium isolates from patients with invasive fusariosis (IF, n= 36) or superficial fusariosis (SF, n= 37), which were obtained at hospitals located in 28 prefectures in Japan between 1998 and 2015, were used for this study. Fusarium isolates were identified using Fusarium- and Fusarium solani species complex (FSSC) -specific real-time PCR and partial DNA sequences of the elongation factor-1 alpha (EF-1α) gene and the nuclear ribosomal internal transcribed spacer (ITS) region. FSSC was predominately isolated from both patients with IF and SF (IF, 77.8% and SF, 67.6%). Distribution of the phylogenetic species of FSSC isolates from patients with IF and SF exhibited different spectra; specifically, F. keratoplasticum (FSSC 2) (25.0%) was the most frequent isolate from patients with IF, whereas F. falciforme (FSSC 3+4) (32.4%) was the most frequent isolate from patients with SF. Fusarium sp. (FSSC 5) was the second most frequent isolate from both patients with IF and SF (IF, 22.2% and SF, 24.3%). Notably, F. petroliphilum (FSSC 1) was isolated only from patients with IF. Each species was isolated from a broad geographic area, and an epidemic was not observed. This is the first epidemiological study of Fusarium species causing IF and SF in Japan.

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 &ldquo, repair template&rdquo, 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.

Fusarium petroliphilum is a recently described species within the Fusarium solani species complex. Some strains of Fusarium species are capable of forming yeast-like structures in tissue as well as in culture through a process known as "adventitious sporulation." Here, we describe the formation of these yeast-like reproductive structures in infected nail tissue obtained from a case of onychomycosis. These structures were also observed in culture grown on RPMI 1640 agar supplemented with 2% glucose. The isolate was resistant to azoles and echinocandins. To the best of our knowledge, this is the first report describing adventitious sporulation in F. petroliphilum and its etiologic role in onychomycosis.

Fusarium species are environmental saprophytic fungi. Among the many Fusarium species, members of the Fusarium solani species complex (FSSC) are the most prevalent and virulent in causing human and animal infections. In this study, we describe the first case of fatal FSSC infection in a black spotted stingray and three concomitant infections in scalloped hammerhead sharks. In the stingray, cutaneous lesions were characterised by ulcers and haemorrhage of the ventral pectoral fin, or 'ray', especially around the head; while cutaneous lesions in the sharks were characterised by ulcers, haemorrhage, as well as white and purulent exudates at the cephalic canals of the cephalofoil and lateral line. Histological sections of the cutaneous lesions revealed slender (1-4 μm in diameter), branching, septate fungal hyphae. Internal transcribed spacer region and 28S nrDNA sequencing of the fungal isolates from the fish showed two isolates were F. keratoplasticum (FSSC 2) and the other two were FSSC 12. Environmental investigation revealed the FSSC strains isolated from water and biofilms in tanks that housed the elasmobranchs were also F. keratoplasticum and FSSC 12. Fusarium is associated with major infections in elasmobranchs and FSSC 12 is an emerging cause of infections in marine animals. DNA sequencing is so far the most reliable method for accurate identification of Fusarium species.

The in vitro activities of amphotericin B in combination with rifampin were assessed against 95 ocular fungal isolates. The interactions between amphotericin B and rifampin at 4, 8, 16, and 32 μg/ml were synergistic for 11.8%, 51.0%, 90.2%, and 94.1%, respectively, of Fusarium solani species complex isolates and for 13.6%, 45.5%, 93.2%, and 95.5%, respectively, of Aspergillus flavus species complex isolates. Antagonism was never observed for the amphotericin B-rifampin combinations.

The Fusarium solani species complex (FSSC) includes important root pathogens of soybean in the United States, but the evolutionary lineages associated with soybean root rot are unknown. A multilocus phylogeny based on 93 isolates from soybean and pea roots from North Dakota and Minnesota revealed that root rot was associated with three known phylogenetic species, FSSC 3 + 4 (=Fusarium falciforme) (3 % of isolates), FSSC 5 (60 %), FSSC 11 (34 %), and one unknown species, FSSC X (2 %). Of these species FSSC 5 and FSSC 3 + 4 are clinically important while FSSC 11 is a plant pathogen. Isolates from FSSC 11 were pathogenic on soybean, dry bean, pea and lentil, and did not grow at 37 °C. However, isolates from FSSC 5 were weakly to non-pathogenic, but grew at 37 °C. Isolates from both FSSC 5 and FSSC 11 were highly resistant to fludioxonil in vitro. This is the first study revealing the pathogenic robustness of FSSC 11 in causing root rot among Fabaceae crops and also the association of clinically important members of the FSSC with roots of a widely grown field crop in the United States.

Members of Fusarium solani species complex (FSSC) are common pathogens of potato, causing dry rot in the west of Iran which involved Hamedan, Kermanshah, Eilam and Kurdistan provinces. Therefore, the objectives in this study were to isolate and identify disease-causing FSSC from infected potato tubers based on the morphological and molecular characteristics. Forty-five isolates of Fusarium were obtained from potato tubers collected from the wet market in different regions of the west of Iran and identified as FSSC through morphological characters. All of the isolates were evaluated for their pathogenicity on healthy potato tubers in the planthouse. The tubers rot symptoms were observed on the 21st day after inoculation of Fusarium isolates on the tubers tested. In the tubers inoculation tests, lesion sizes were quite variable; therefore, the measurement was done to compare the depth and width of lesion expansion among the isolates. Based on the sequence data from translation elongation factor (EF-lα) gene and internal transcript spacer (ITS) regions analysis, all of the selected FSSC isolates were divided into two major groups. This is the first report on molecular identification of FSSC strains isolated from potato tubers in Iran and Fusarium falciforme was reported for the first time in Iran.

Aims: The objective of this work was to design an amplified fragment length polymorphism (AFLP)-derived specific primer for the detection of Fusarium solani aetiological agent of peanut brown root rot (PBRR) in plant material and soil. Methods and Results: Specific primers for the detection of the pathogen were designed based on an amplified region using AFLPs. The banding patterns by AFLPs showed that isolates from diseased roots were clearly distinguishable from others members of the F. solani species complex. Many bands were specific to F. solani PBRR, one of these fragments was selected and sequenced. Sequence obtained was used to develop specific PCR primers for the identification of pathogen in pure culture and in plant material and soil. Primer pair FS1/FS2 amplified a single DNA product of 175 bp. Other fungal isolates occurring in soil, included F. solani non-PBRR, were not detected by these specific primers. The assay was effective for the detection of pathogen from diseased root and infected soils. Conclusions: The designed primers for F. solani causing PBRR can be used in a PCR diagnostic protocol to rapidly and reliably detect and identify this pathogen. Significance and Impact of the Study: These diagnostic PCR primers will aid the detection of F. solani causing PBRR in diseased root and natural infected soils. The method developed could be a helpful tool for epidemiological studies and to avoid the spread of this serious disease in new areas. Fil: Casasnovas, F.. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina Fil: Fantini, Emanuel Nicolás. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Palazzini, Juan Manuel. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Giaj Merlera, Guillermo. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Chulze, Sofia Noemi. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Reynoso, Maria Marta. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Torres, Adriana Mabel. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina

3 pages, 1 table, 18 references. The collaboration of the Lascaux restoration team is highly appreciated. We thank Marisa Chelius for valuable comments on the manuscript., The LascauxCave was discovered in 1940, and by 1960 it had received up to 1800 daily visitors. In 1963, the cave was closed and in 2001 it was invaded by a Fusarium solani species complex which was treated for four years with benzalkonium chloride. However, LascauxCavebacteria have only been poorly investigated. Here we show that the cave is now a reservoir of potential pathogenic bacteria and protozoa which can be found in outbreaks linked to air-conditioning systems and cooling towers in community hospitals and public buildings., The first author and the research project “Microbiology-Microclimate” were supported by the Ministry of Culture and Communication, France. Authors acknowledge support from the Ministry of Culture and Communication and from project CGL2006-07424/BOS, Spain.

In two recent studies, clinical isolates in the Fusarium solani species complex (FSSC) were sequenced; one of the most common lineages was FSSC Group 1 (FSSC 1), a phylogenetic species that is synonymous with F. solani f. sp. cucurbitae race 2, a pathogen of cucurbit fruits. FSSC 1 was also identified in sink and shower drains in two hospitals. The environmental sources of FSSC 1 are important for understanding the epidemiology of both human and plant diseases caused by this organism. FSSC 1 was detected in sewage influent at all six tested urban wastewater treatment plants (WWTPs) in California with a concentration ranging from 75 to 413 colony-forming units (cfu) l(-1), a mean of 246 +/- 52 cfu l(-1) and a median of 254 cfu l(-1). During the treatment process, the concentration of FSSC 1 in the solid and liquid fractions diminished. FSSC 1 was detected in five and six of 14 community shower drains by culturing and polymerase chain reaction, respectively, whereas FSSC DNA was detected in all drains. FSSC accounted for 17 +/- 6% (n = 14) of the total fungal DNA in the drains. FSSC 1 was rarely isolated from post-harvest cucurbit fruits and was not found in cucurbit fields in California.