RESUMO
Onychomycosis is a common chronic nail disorder where dermatophytes are the predominant pathogens. However, non-dermatophytic moulds like Aspergillus can also be implicated as the causative agents. Herein, we report a rare case of onychomycosis due to Emericella quadrilineata ( Aspergillus tetrazonus) in an apparently immunocompetent host.
Assuntos
Emericella/classificação , Emericella/isolamento & purificação , Onicomicose/diagnóstico , Onicomicose/patologia , Emericella/citologia , Humanos , Masculino , Técnicas Microbiológicas , Microscopia , Pessoa de Meia-Idade , Onicomicose/microbiologiaRESUMO
A 78-year-old male who was undergoing prolonged glucocorticoid treatment experienced cough and expectoration for 2 weeks. Galactomannan antigen analysis and a chest computed tomography (CT) scan suggested a diagnosis of invasive pulmonary aspergillosis. DNA sequencing indicated that Emericella nidulans var. echinulata was the causative agent. A combination of voriconazole and micafungin successfully treated the illness.
Assuntos
Antifúngicos/administração & dosagem , Equinocandinas/administração & dosagem , Emericella/isolamento & purificação , Aspergilose Pulmonar Invasiva/diagnóstico , Aspergilose Pulmonar Invasiva/tratamento farmacológico , Lipopeptídeos/administração & dosagem , Pirimidinas/administração & dosagem , Triazóis/administração & dosagem , Idoso , DNA Fúngico/química , DNA Fúngico/genética , Emericella/classificação , Emericella/genética , Exsudatos e Transudatos/microbiologia , Galactose/análogos & derivados , Glucocorticoides/administração & dosagem , Glucocorticoides/efeitos adversos , Humanos , Hospedeiro Imunocomprometido , Aspergilose Pulmonar Invasiva/microbiologia , Masculino , Mananas/análise , Micafungina , Microscopia , Dados de Sequência Molecular , Análise de Sequência de DNA , Resultado do Tratamento , VoriconazolRESUMO
OBJECTIVE: To characterize, identify and investigate the anticancer properties of two new soil fungal isolates, Emericella nidulans and Fusarium solani isolated from Wady El-Natron in Egypt against colon cancer Caco-2 (ATCC) cell line. METHODS: Soil sample was cultured and two strains were chosen for morphological and phenotypical characterization. Partial sequences of the 18s rRNA gene and the internal transcribed spacer region ITS of the two isolates were amplified by PCR. Phylogenetic tree construction and analysis of the resulted multiple sequences from the two fugal isolates were also carried out. In vitro anticancer activity of the two strains was done against colon Caco-2 cancer cell line. Reverse transcription - PCR was carried out to detect level of expression of p53 in Caco-2 cell line. RESULTS: HF.1 displayed morphological and genotypic characteristics most similar to that of Fusarium solani while HF.2 was most similar to Emericella nidulans with high similarity of 99% and 97% respectively. The multiple sequence alignment of the two fungal isolates showed that, the maximum identical conserved domains in the 18s rRNA genes were identified with the nucleotide regions of 51st to 399th base pairs, 88th to 525th base pairs respectively. While those in the ITS genes were identified with the nucleotide regions of 88th to 463rd and 51st to 274th. The two isolates showed IC50 value with (6.24±5.21) and (9.84±0.36) µg/mL) concentrations respectively at 28h. Reverse transcription - PCR indicated that these cells showed high level of expression for p53 mRNA. CONCLUSIONS: The morphology and molecular analysis identified HF.1 and HF.2 to be Fusarium solani and Emericella nidulans; new isolates of anticancer producing fungi from Wady El-Natroon city in Egypt. Treatment with the two isolates caused P53 expression in Caco-2 cell line. These two isolates can be used as an anticancer agents.
Assuntos
Antineoplásicos/farmacologia , Misturas Complexas/farmacologia , Emericella/isolamento & purificação , Fusarium/isolamento & purificação , Microbiologia do Solo , Antineoplásicos/química , Células CACO-2 , Misturas Complexas/química , Egito , Emericella/química , Emericella/classificação , Emericella/genética , Fusarium/química , Fusarium/classificação , Fusarium/genética , Expressão Gênica/efeitos dos fármacos , Humanos , Filogenia , Proteína Supressora de Tumor p53/genética , Proteína Supressora de Tumor p53/metabolismoRESUMO
Polyphasic characterization of the echinocandin B producer Aspergillus nidulans var. roseus ATCC 58397 strain was carried out to elucidate its taxonomical status. According to its carbon source utilization and secondary metabolite spectrum as well as the partial ß-tubulin, calmodulin, and γ-actin gene sequences, A. nidulans var. roseus belongs to the Emericella rugulosa species. Auxotroph mutants of A. nidulans var. roseus ATCC 58397 and E. rugulosa CBS 171.71 and CBS 133.60 formed stable heterokaryons on minimal medium with several A. nidulans strains, and in the case of A. nidulans var. roseus, even cleistothecia were developed.
Assuntos
Actinas/genética , Aspergillus nidulans/genética , Calmodulina/genética , Equinocandinas/biossíntese , Emericella/genética , Proteínas Fúngicas/biossíntese , Tubulina (Proteína)/genética , Actinas/química , Actinas/metabolismo , Aspergillus nidulans/classificação , Aspergillus nidulans/metabolismo , Calmodulina/química , Calmodulina/metabolismo , Cruzamentos Genéticos , Emericella/classificação , Emericella/metabolismo , Fermentação , Proteínas Fúngicas/química , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Técnicas de Tipagem Micológica , Filogenia , Reação em Cadeia da Polimerase , Análise de Sequência de DNA , Esterigmatocistina/biossíntese , Tubulina (Proteína)/química , Tubulina (Proteína)/metabolismoRESUMO
The mode of reproduction of the soil ascomycetous fungus Emericella nidulans of Israeli populations was studied using 15 microsatellite (simple sequence repeats or SSR) trinucleotide markers. The study was performed in three canyons: two located in the northern part of Israel (Mount Carmel and western Upper Galilee) and one in the southern Negev desert. In each canyon, E. nidulans strains were isolated from the opposite slopes and (in the desert canyon) the valley bottom. Testing the reproductive structure of the populations indicated the presence of sexuality in the northern population and predominant clonality in the desert population. The predominantly clonal character of the desert population of E. nidulans was explained by the assumption that for relevant multilocus systems of a fungus, only several haplotypes can survive in the rather constant, extremely stressful desert conditions. Additionally, the very low density of E. nidulans populations in the soil of the desert canyon, which reduces the probability of finding a sexual partner, might favour predominant clonality via selfing. Increasing sexuality in E. nidulans populations on the north-facing slopes of the northern canyons may be a result of biotic stress (pressure of competitive fungal species), due to the more mild ecological conditions in these canyons.
Assuntos
Emericella/genética , Adaptação Fisiológica , Biodiversidade , Emericella/classificação , Emericella/fisiologia , Meio Ambiente , Evolução Molecular , Genes Fúngicos , Genética Populacional , Israel , Desequilíbrio de Ligação , Repetições de MicrossatélitesRESUMO
Four new species of Emericella, E. discophora, E. filifera, E. olivicola and E. stella-maris, are proposed. Their new taxonomic status was determined applying a polyphasic taxonomic approach using phenotypic (morphology and extrolite profiles) and molecular (sequences of ITS, beta-tubulin and calmodulin genes) characters. Ascospores of E. stella-maris and E. olivicola have star-shape equatorial crests, those of E. filifera form long appendages that emerge radially from narrow stellate crests and those of E. discophora produce wide and entire, nonstellate equatorial crests. E. stella-maris originated from leaf litter in Tunisia and E. filifera from raisins in Argentina, and both of them also were found in hypersaline water of a saltern in Slovenia. E. olivicola was isolated from olives in Italy and E. discophora from soil in Spain. All listed species possess distinct extrolite profiles: E. stella-maris produced arugosin E, shamixanthone and the yet unelucidated metabolites glia 1-3; E. filifera produced shamixanthone and varitriols; E. discophora produced sterigmatocystin and versicolorins; E. olivicola produced numerous extrolites such as arugosin E, siderin, shamixanthone, sterigmatocystin, terrein, varitriols and aflatoxin B1, of which the latter was detected only in one of the two strains.
Assuntos
Emericella/classificação , Emericella/isolamento & purificação , Água do Mar/microbiologia , Microbiologia do Solo , DNA Fúngico/genética , DNA Espaçador Ribossômico/genética , Emericella/genética , Emericella/metabolismo , Europa (Continente) , Proteínas Fúngicas/genética , Dados de Sequência Molecular , Micotoxinas/metabolismo , Filogenia , Esporos Fúngicos/citologiaRESUMO
We noted a cluster of 4 cases of infection or colonization by Emericella spp., identified by sequence-based analysis as E. quadrilineata. Sequence-based analysis of an international collection of 33 Emericella isolates identified 12 as E. nidulans, all 12 of which had previously been identified by morphologic methods as E. nidulans. For 12 isolates classified as E. quadrilineata, only 6 had been previously identified accordingly. E. nidulans was less susceptible than E. quadrilineata to amphotericin B (median MICs 2.5 and 0.5 mg/L, respectively, p<0.05); E. quadrilineata was less susceptible than E. nidulans to caspofungin (median MICs, 1.83 and 0.32 mg/L, respectively, p<0.05). These data indicate that sequence-based identification is more accurate than morphologic examination for identifying Emericella spp. and that correct species demarcation and in vitro susceptibility testing may affect patient management.
Assuntos
Emericella/isolamento & purificação , Micoses/microbiologia , Adulto , Antifúngicos/farmacologia , Aspergilose/microbiologia , Composição de Bases , Criança , DNA Fúngico , DNA Intergênico/genética , Emericella/classificação , Emericella/efeitos dos fármacos , Emericella/genética , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , FilogeniaRESUMO
Most aspergilli that produce aflatoxin are members of Aspergillus section Flavi, however isolates of several Aspergillus species not closely related to section Flavi also have been found to produce aflatoxin. Two of the species, Aspergillus ochraceoroseus and an undescribed Aspergillus species SRRC 1468, are morphologically similar to members of Aspergillus section Circumdati. The other species have Emericella teleomorphs (Em. astellata and an undescribed Emericella species SRRC 2520) and are morphologically distinctive in having ascospores with large flanges. All these aflatoxin-producing isolates were from tropical zones near oceans, and none of them grew on artificial media at 37 C. Aflatoxins and sterigmatocystin production were quantified by high-pressure liquid chromatography (HPLC) and confirmed by HPLC-mass spectrometry (LC-MS) detection. Phylogenetic analyses were conducted on these four species using A. parasiticus and Em. nidulans, (which produce aflatoxin and the aflatoxin precursor sterigmatocystin, respectively) for comparison. Two aflatoxin/sterigmatocystin biosynthesis genes and the beta tubulin gene were used in the analyses. Results showed that of the new aflatoxin-producers, Aspergillus SRRC 1468 forms a strongly supported clade with A. ochraceoroseus as does Emericella SRRC 2520 with Em. astellata SRRC 503 and 512.
Assuntos
Aflatoxinas/análise , Aspergillus/citologia , Aspergillus/genética , Emericella/citologia , Emericella/genética , Aflatoxinas/genética , Aspergillus/química , Aspergillus/classificação , Cromatografia Líquida de Alta Pressão , DNA Fúngico/química , DNA Fúngico/genética , Emericella/química , Emericella/classificação , Proteínas Fúngicas/genética , Espectrometria de Massas , Dados de Sequência Molecular , Filogenia , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos , Esterigmatocistina/análise , Tubulina (Proteína)/genéticaRESUMO
Emericella venezuelensis is a new species, differing from two other species with stellate ascospores, E. variecolor and E. pluriseminata, by triangular flaps on the convex sides of the ascospores, and further from E. variecolor by producing an Aspergillus anamorph only on unconventional growth media. The three species also differ in their profiles of extrolites (secondary metabolites). Emericella venezuelensis produces aflatoxin B1, sterigmatocystin, and terrein and compounds with chromophores of the shamixanthone, emerin and desertorin type of compounds. E. variecolor produces asteltoxin, shamixanthone, asperthecin, and terrein, in addition to metabolites unequivocally recorded in the literature or tentatively identified here as astellolide A & B, andibenin A, B, C, andilesin A, B, C, anditomin, astellatol, stellatic acid, stellatin, tajixanthone, radixanthone, najamxanthone, ajamxanthone, variecoxanthone A, B, C, isoemericellin, kojic acid, varitriol, varioxiran, dihydroterrein, 7-hydroxyemodin, avariquinone and stromemycin. E. pluriseminata produces several unknown specific extrolites. E. venezuelensis is the first organism of marine origin reported to produce aflatoxin. Aflatoxin production by E. venezuelensis makes this species an attractive model organism for the study of the regulation of this important type of carcinogenic mycotoxins in combination with the knowledge on sterigmatocystin production by E. nidulans, soon to be whole genome sequenced. The isolates were also analyzed cladistically using partial sequences of the beta-tubulin gene. Since three species of Emericella have stellate ascospores, and the type material of E. variecolor is equivocal, this species is epitypified with CBS 598.65. Emericella species normally do not appear to cause problems for food safety, as they are most often found in litter and soil.
Assuntos
Aflatoxina B1/biossíntese , Emericella/classificação , Emericella/isolamento & purificação , Poríferos/microbiologia , Esterigmatocistina/biossíntese , Animais , Álcoois Benzílicos/análise , Álcoois Benzílicos/isolamento & purificação , Ciclopentanos/análise , Ciclopentanos/isolamento & purificação , DNA Fúngico/química , DNA Fúngico/isolamento & purificação , Emericella/citologia , Emericella/metabolismo , Furanos/análise , Furanos/isolamento & purificação , Genes Fúngicos , Proteínas de Membrana/análise , Proteínas de Membrana/isolamento & purificação , Dados de Sequência Molecular , Proteínas Nucleares , Filogenia , Pironas/análise , Pironas/isolamento & purificação , Água do Mar/microbiologia , Análise de Sequência de DNA , Esporos Fúngicos/citologia , Timopoietinas/análise , Timopoietinas/isolamento & purificação , Tubulina (Proteína)/genética , Microbiologia da Água , Xantonas/análise , Xantonas/isolamento & purificaçãoRESUMO
AIMS: To report on aflatoxin B(1) and B(2) production from a species of Emericella. METHODS AND RESULTS: Aflatoxins and sterigmatocystin were determined by high-pressure liquid chromatography (HPLC) with diode array detection and confirmed by HPLC with mass spectrometry detection. Among 30 known species of Emericella only one species produced aflatoxin. Strains originating from the same geographical source material had different patterns of aflatoxin and sterigmatocystin production on different media, indicating that epigenetic factors may be involved in the regulation of aflatoxin production. However, two cultures from the same original genet were very similar. CONCLUSIONS: Emericella astellata can produce small amounts of sterigmatocystin and aflatoxin B(1) and B(2). SIGNIFICANCE AND IMPACT OF THE STUDY: Emericella has been used extensively in genetic studies and therefore the isolates producing aflatoxin can be used to elucidate the genetic, evolutionary and maybe ecological role of aflatoxins using molecular genetic methods.
Assuntos
Aflatoxinas/biossíntese , Emericella/metabolismo , Esterigmatocistina/biossíntese , Aflatoxina B1/biossíntese , Cromatografia Líquida , Meios de Cultura , Emericella/classificação , Emericella/crescimento & desenvolvimento , Espectrometria de MassasRESUMO
Nondermatophytic fungi are increasingly being reported as etiological agents of onychomycosis. We describe here a case of hand nail infection caused by Emericella quadrilineata (anamorph Aspergillus tetrazonus), a species not so far known to be an etiological agent of onychomycosis.