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1.
J Laryngol Otol ; 134(7): 632-635, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32686637

RESUMO

BACKGROUND: Invasive fungal rhinosinusitis is associated with high morbidity and mortality. Rapid pathogen identification is mandatory, but fresh tissue is not always available. A polymerase chain reaction method was designed in order to detect fungi in formalin-fixed paraffin-embedded samples. This was applied to a retrospective series of tissue biopsies from Thai patients with invasive fungal rhinosinusitis. METHODS: Tissue blocks from 64 cases yielded adequate DNA. Three sequential polymerase chain reaction were performed: ZP3 (housekeeping gene) and panfungal polymerase chain reactions, and a differentiating polymerase chain reaction based on the 5.8s ribosomal RNA and internal transcribed spacer 2 regions. The polymerase chain reaction products were then sequenced. RESULTS: Polymerase chain reaction identified a fungal pathogen in 20 of 64 cases (31 per cent). Aspergillus species was the most common cause of invasive fungal rhinosinusitis (nine cases). Other causes included candida (n = 4), cladosporium (n = 4), mucor (n = 1), alternaria (n = 1) and dendryphiella (n = 1) species. CONCLUSION: Polymerase chain reaction can provide rapid identification of fungal pathogens in paraffin-embedded tissue, enabling prompt treatment of invasive fungal rhinosinusitis.


Assuntos
Micoses/microbiologia , Reação em Cadeia da Polimerase/métodos , Rinite/microbiologia , Sinusite/microbiologia , Aspergillus/genética , Biópsia , Candida/genética , Criança , Pré-Escolar , Cladosporium/genética , DNA Fúngico/genética , Humanos , Lactente , Inclusão em Parafina , RNA Ribossômico 5,8S/genética , Estudos Retrospectivos , Rinite/patologia , Sinusite/patologia
2.
Int J Syst Evol Microbiol ; 70(8): 4458-4469, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32674752

RESUMO

Four new yeast species belonging to the genus Apiotrichum and two new yeast species belonging to Cutaneotrichosporon are described for strains isolated from guano samples from bat-inhabited caves in Japan. In 2005, we reported these isolates as Trichosporon species based on sequence analyses of the D1/D2 domain of large subunit (LSU) rRNA genes according to available basidiomycetous yeast classification criteria; however, to date, they have not been officially published as new species with descriptions. Their phylogenetic positions have been reanalysed based on comparison of internal transcribed spacer (ITS) region sequences (including the 5.8S rRNA gene) and the D1/D2 domain of the LSU rRNA gene with those of known species; we confirmed clear separation from previously described species. Physiological and biochemical properties of the isolates also suggest their distinctiveness. Therefore, we describe Apiotrichum akiyoshidainum (holotype JCM 12595T), Apiotrichum chiropterorum (JCM 12594T), Apiotrichum coprophilum (JCM 12596T), Apiotrichum otae (JCM 12593T), Cutaneotrichosporon cavernicola (JCM 12590T) and Cutaneotrichosporon middelhovenii (JCM 12592T) as new species. C. cavernicola showed particularly distinctive morphology including large inflated anomalous cells on the hyphae and germination from the cells, although clear clamp connections on the hyphae were not confirmed. Further study is needed to elucidate the morph of this species.


Assuntos
Basidiomycota/classificação , Quirópteros/microbiologia , Fezes/microbiologia , Filogenia , Animais , Basidiomycota/isolamento & purificação , DNA Fúngico/genética , DNA Espaçador Ribossômico/genética , Japão , Técnicas de Tipagem Micológica , RNA Ribossômico 5,8S/genética , Análise de Sequência de DNA
3.
Int J Food Microbiol ; 312: 108373, 2020 Jan 02.
Artigo em Inglês | MEDLINE | ID: mdl-31654841

RESUMO

The use of non-Saccharomyces yeast in conjunction with Saccharomyces cerevisiae in wine fermentation is a growing trend in the wine industry. Non-Saccharomyces, through their distinctive production of secondary metabolites, have the potential to positively contribute to wine sensory profile. To discover new candidate strains for development as starter cultures, indigenous non-Saccharomyces were isolated from un-inoculated fermenting Shiraz musts from a South Australian vineyard (McLaren Vale wine region) and characterised. Among the 77 isolates, 7 species belonging to 5 genera (Kazachstania, Aureobasidium, Meyerozyma, Wickerhamomyces and Torulaspora) were identified by sequencing the internal transcribed spacer regions of the 5.8S rRNA gene (ITS1-5.8S-ITS2 region). The indigenous isolates were evaluated for oenological properties, namely, ethanol tolerance, enzyme activity, and H2S production. To determine their potential industrial use as starter cultures, representative isolates of each species were assessed in a sterile chemically defined grape juice and Viognier grape juice to evaluate their contribution to fermentation kinetics and production of key metabolites, including volatile compounds.


Assuntos
Saccharomycetales/genética , Saccharomycetales/metabolismo , Vitis/metabolismo , Vinho/microbiologia , Leveduras/metabolismo , Austrália , DNA Intergênico/genética , Fazendas , Fermentação , RNA Ribossômico 5,8S/genética , Saccharomycetales/classificação , Saccharomycetales/isolamento & purificação , Austrália do Sul , Vinho/análise
4.
Mycologia ; 111(5): 857-870, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31414967

RESUMO

In this paper, species of the genus Morchella are investigated in China. Based on morphological characteristics and molecular phylogenetic analyses of the nuc rDNA internal transcribed spacer region ITS1-5.8S-ITS2 (ITS) and the combined data set ITS + nuclear large subunit rDNA (28S) + the translation elongation factor 1-α (TEF1) gene + RNA polymerase II first largest subunit (RPB1) + RNA polymerase II second largest subunit (RPB2), six new phylogenetic species are illustrated and described: M. clivicola, M. confusa, M. odonnellii, M. owneri, M. yangii, and M. yishuica. Furthermore, two new record species, M. dunensis and M. palazonii, which were only known in Europe, are now reported for the first time from Asia. New species of morels will provide additional information on species diversity and genetic resource candidates for improving the cultivation of this economically important fungus.


Assuntos
Ascomicetos/classificação , Ascomicetos/isolamento & purificação , Filogenia , Ascomicetos/genética , Ascomicetos/crescimento & desenvolvimento , China , Análise por Conglomerados , DNA Fúngico/química , DNA Fúngico/genética , DNA Ribossômico/química , DNA Ribossômico/genética , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , Genes de RNAr , Fator 1 de Elongação de Peptídeos/genética , RNA Polimerase I/genética , RNA Polimerase II/genética , RNA Fúngico/genética , RNA Ribossômico 28S/genética , RNA Ribossômico 5,8S/genética , Análise de Sequência de DNA
5.
PLoS Negl Trop Dis ; 13(8): e0007631, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31425540

RESUMO

OBJECTIVE: Where human African trypanosomiasis (HAT) patients are seen, failure to microscopically diagnose infections by Trypanosoma brucei gambiense in blood smears and/or cerebrospinal fluid (CSF) in the critical early stages of the disease is the single most important factor in treatment failure, a result of delayed treatment onset or its absence. We hypothesized that the enhanced sensitivity of detergent-enhanced loop-mediated isothermal amplification (LAMP) will allow for point of care (POC) detection of African trypanosomes in the CSF of HAT patients where the probability for detecting a single parasite or parasite DNA molecule in 1 µL of CSF sample is negligible by current methods. METHODOLOGY: We used LAMP targeting the multicopy pan-T. brucei repetitive insertion mobile element (RIME LAMP) and the Trypanosoma brucei gambiense 5.8S rRNA-internal transcribed spacer 2 gene (TBG1 LAMP). We tested 1 µL out of 20 µL sham or Triton X-100 treated CSFs from 73 stage-1 and 77 stage-2 HAT patients from the Central African Republic and 100 CSF negative controls. RESULTS: Under sham conditions, parasite DNA was detected by RIME and TBG1 LAMP in 1.4% of the stage-1 and stage-2 gambiense HAT CSF samples tested. After sample incubation with detergent, the number of LAMP parasite positive stage-2 CSF's increased to 26%, a value which included the 2 of the 4 CSF samples where trypanosomes were identified microscopically. Unexpected was the 41% increase in parasite positive stage-1 CSF's detected by LAMP. Cohen's kappa coefficients for RIME versus TBG1 LAMP of 0.92 (95%CI: 0.82-1.00) for stage-1 and 0.90 (95%CI: 0.80-1.00) for stage-2 reflected a high level of agreement between the data sets indicating that the results were not due to amplicon contamination, data confirmed in χ2 tests (p<0.001) and Fisher's exact probability test (p = 4.7e-13). CONCLUSION: This study detected genomic trypanosome DNA in the CSF independent of the HAT stage and may be consistent with early CNS entry and other scenarios that identify critical knowledge gaps for future studies. Detergent-enhanced LAMP could be applicable for non-invasive African trypanosome detection in human skin and saliva or as an epidemiologic tool for the determination of human (or animal) African trypanosome prevalence in areas where chronically low parasitemias are present.


Assuntos
Líquido Cefalorraquidiano/parasitologia , Técnicas de Diagnóstico Molecular/métodos , Técnicas de Amplificação de Ácido Nucleico/métodos , Índice de Gravidade de Doença , Trypanosoma/isolamento & purificação , Tripanossomíase Africana/diagnóstico , Tripanossomíase Africana/patologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , República Centro-Africana , Criança , Pré-Escolar , DNA de Protozoário/genética , DNA Ribossômico/química , DNA Ribossômico/genética , DNA Espaçador Ribossômico/genética , Detergentes/metabolismo , Feminino , Humanos , Lactente , Recém-Nascido , Masculino , Pessoa de Meia-Idade , RNA Ribossômico 5,8S/genética , Sensibilidade e Especificidade , Trypanosoma/genética , Adulto Jovem
6.
Mycologia ; 111(5): 758-771, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31408397

RESUMO

To meet a global demand for timber, tree plantations were established in South America during the first half of the 20th century. Extensive plantings of non-native species now are found in Brazil, Chile, Argentina, and Uruguay. In Colombia, miscellaneous plantations were established in the 1950s, during a period of intensive local logging, when policies to limit deforestation in native Quercus humboldtii forests were established. One unforeseen consequence of planting non-native trees was the simultaneous introduction and subsequent persistence of ectomycorrhizal fungi. We sought to document the origins and spread of the introduced Amanita muscaria found in Colombian plantations of the Mexican species Pinus patula, North American species P. taeda, and Australian species Acacia melanoxylon and Eucalyptus globulus. In Colombia, Amanita muscaria is establishing a novel association with native Q. humboldtii and has spread to local Q. humboldtii forests. According to a Bayesian phylogeny and haplotype analysis based on the nuclear rDNA internal transcribed spacer region ITS1-5.8-ITS2 (ITS barcode), A. muscaria individuals found in four exotic plant species, and those colonizing Q. humboldtii roots, have a Eurasian origin and belong to two Eurasian haplotypes. This is the first time the spread of an introduced mutualist fungus into native Colombian Q. humboldtii forests is reported. To arrest its spread, we suggest the use of local inocula made up of native fungi, instead of inocula of introduced fungi.


Assuntos
Amanita/crescimento & desenvolvimento , Amanita/isolamento & purificação , Especificidade de Hospedeiro , Quercus/microbiologia , Acacia/microbiologia , Amanita/genética , Análise por Conglomerados , Colômbia , DNA Fúngico/química , DNA Fúngico/genética , DNA Ribossômico/química , DNA Ribossômico/genética , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , Eucalyptus/microbiologia , Florestas , Filogenia , Pinus/microbiologia , RNA Ribossômico 5,8S/genética , Análise de Sequência de DNA
7.
Food Sci Technol Int ; 25(8): 671-679, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31272221

RESUMO

Spoilage caused by yeasts is a constant, widespread problem in the beverage industry that can result in major economic losses. Fruit juices provide an environment that allows the proliferation of yeast. Some factories in South Africa are not equipped with laboratory facilities to identify spoilage yeasts and outsourcing becomes a prolonged process which obstructs corrective action planning. This study aimed to establish yeast diversity and apply a rapid method for preliminary identification of spoilage yeasts associated with a small-scale fruit juice bottling factory. Yeast population in the factory was determined by isolation from the production environment, process equipment and spoiled products. PCR-RFLP analysis targeting the 5.8S-ITS region and D1/D2 sequencing was used for identification. A total of 207 yeasts belonging to 10 different genera (Candida, Lodderomyces, Wickerhamomyces, Yarrowia, Zygosaccharomyces, Zygoascus, Cryptococcus, Filobasidium, Rhodotorula/Cystobasidium and Trichosporon) were isolated and identified from the production environment and processing equipment. Candida intermedia, C. parapsilosis and Lodderomyces elongisporus were widely distributed in the factory. Zygosaccharomyces bailii, Z. bisporus, Zygoascus hellenicus and Saccharomyces cerevisiae were isolated from the spoiled products. The data provided a yeast control panel that was used successfully to identify unknown yeasts in spoiled products from this factory using polymerase chain reaction-restriction length polymorphism (PCR-RFLP) comparative analysis.


Assuntos
Contaminação de Alimentos/análise , Sucos de Frutas e Vegetais/microbiologia , Reação em Cadeia da Polimerase/métodos , Polimorfismo de Fragmento de Restrição , Leveduras/classificação , Leveduras/isolamento & purificação , Biodiversidade , DNA Fúngico , Manipulação de Alimentos , Técnicas de Tipagem Micológica , RNA Ribossômico/análise , RNA Ribossômico/genética , RNA Ribossômico 5,8S/análise , RNA Ribossômico 5,8S/genética , Análise de Sequência , Leveduras/genética , Leveduras/crescimento & desenvolvimento
8.
Mycologia ; 111(4): 660-675, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31150307

RESUMO

Species of Phaeochorella are biotrophic leaf parasites with a tropical distribution, traditionally accepted in the family Phyllachoraceae, Phyllachorales in classifications based on morphological characters. Phylogenetic evidence presented here resolves the relationship of Phaeochorella within the Sordariomycetes, based on a multilocus analysis of partial nuc rDNA large subunit (28S) and internal transcribed spacers (ITS1-5.8S-ITS2 = ITS), the DNA-directed RNA polymerase II second largest subunit (RPB2), and the translation elongation factor 1-α (TEF1-α) gene. Phylogenetic analyses indicate that Phaeochorella belongs to the Diaporthales rather than the Phyllachorales. Phaeochorella parinarii, the type species of the genus, present on native hosts from the Brazilian Cerrado, forms a unique clade with a species of Phaeoappendicospora with high support. Thus, a new family, Phaeochorellaceae, Diaporthales, including both genera, is herein proposed. With the exception of P. parinarii and P. zonata, all other species in Phaeochorella (P. artocarpi, P. ciliata, P. machaerii) were excluded from the genus.


Assuntos
Phyllachorales/classificação , Phyllachorales/isolamento & purificação , Filogenia , Ascomicetos/classificação , Ascomicetos/genética , Ascomicetos/isolamento & purificação , Brasil , DNA Fúngico/genética , Phyllachorales/genética , RNA Ribossômico 28S/genética , RNA Ribossômico 5,8S/genética
9.
Antonie Van Leeuwenhoek ; 112(10): 1501-1521, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31140027

RESUMO

Fungi under the order Ophiostomatales (Ascomycota) are known to associate with various species of bark beetles (Coleoptera: Curculionidae: Scolytinae). In addition this group of fungi contains many taxa that can impart blue-stain on sapwood and some are important tree pathogens. A recent survey that focussed on the diversity of the Ophiostomatales in the forest ecosystems of the Czech Republic and Poland uncovered four putative new species. Phylogenetic analyses of four gene regions (ITS1-5.8S-ITS2 region, ß-tubulin, calmodulin, and translation elongation factor 1-α) indicated that these four species are members of the genus Ophiostoma. All four newly described species can be distinguished from each other and from closely related species based on DNA sequence comparisons, morphological characters, growth rates, and their insect associations. Based on this study four new taxa can be circumscribed and the following names are provided: Ophiostoma pityokteinis sp. nov., Ophiostoma rufum sp. nov., Ophiostoma solheimii sp. nov., and Ophiostoma taphrorychi sp. nov. O. rufum sp. nov. is a member of the Ophiostoma piceae species complex, while O. pityokteinis sp. nov. resides in a discrete lineage within Ophiostoma s. stricto. O. taphrorychi sp. nov. together with O. distortum formed a well-supported clade in Ophiostoma s. stricto close to O. pityokteinis sp. nov. O. solheimii sp. nov. groups within a currently undefined lineage A, which also includes Ophiostoma grandicarpum and Ophiostoma microsporum. This study highlights the need for more intensive surveys that should include additional countries of Central Europe, insect vectors and host tree species in order to elucidate Ophiostoma species diversity in this region.


Assuntos
Ophiostoma/classificação , Ophiostoma/isolamento & purificação , Filogenia , Gorgulhos/microbiologia , Animais , Calmodulina/genética , República Tcheca , DNA Espaçador Ribossômico/genética , Técnicas de Tipagem Micológica , Ophiostoma/genética , Ophiostoma/fisiologia , Fator 1 de Elongação de Peptídeos/genética , Polônia , RNA Ribossômico 5,8S/genética , Análise de Sequência de DNA , Traqueófitas/parasitologia , Tubulina (Proteína)/genética , Gorgulhos/crescimento & desenvolvimento , Madeira/parasitologia
10.
Mycologia ; 111(3): 456-465, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30985272

RESUMO

Stevia (Stevia rebaudiana) is an emerging perennial crop in the southeastern United States. A Septoria leaf spot disease of stevia was first identified on field plantings in Japan in 1978. The pathogen was named Septoria steviae based on a morphological characterization. In 2015, a species of Septoria with morphological characters of S. steviae was isolated from field and greenhouse-grown stevia plants with leaf spot symptoms in North Carolina. In this study, 12 isolates obtained from diseased stevia plants in 2015 and 2016 were characterized and compared with reference strains of S. steviae. Comparisons were based on conidial and pycnidial morphology and multilocus sequence analysis of actin (ACT), ß-tubulin (BT), calmodulin (CAL), nuc rDNA internal transcribed spacers (ITS1-5.8S-ITS2 = ITS), nuc rDNA 28S subunit (28S), RNA polymerase II second largest subunit (RPB2), and translation elongation factor-1α (TEF1). Measurements of conidia and pycnidia from symptomatic field leaves and 12 pure cultures grown on nutrient medium were consistent with those previously reported for ex-type strains of S. steviae. North Carolina strains formed a well-supported monophyletic group with ex-type strains of S. steviae. This study represents the first genetic characterization of S. steviae in the United States and provides an experimental framework to elucidate the genetic diversity and disease ecology of field populations of S. steviae.


Assuntos
Ascomicetos/genética , Filogenia , Doenças das Plantas/microbiologia , Folhas de Planta/microbiologia , Stevia/microbiologia , Ascomicetos/patogenicidade , DNA Fúngico/genética , DNA Espaçador Ribossômico/genética , North Carolina , RNA Ribossômico 5,8S/genética , Análise de Sequência de DNA
11.
Mycologia ; 111(2): 299-318, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30924725

RESUMO

Aquanectria and Gliocladiopsis are two closely related genera of Hypocreales. They are also morphologically similar, forming hyaline, penicillate conidiophores and hyaline, straight to sinuous, 0-1-septate phialoconidia. During a revision of gliocladiopsis-like isolates originating from rain forest areas of South America (Ecuador, French Guiana) and Southeast Asia (Singapore), multilocus phylogenetic inferences, based on DNA sequences encoding partial ß-tubulin (TUB2), translation elongation factor 1-α (TEF1- α), histone H3 (HIS3) genes and the nuc rDNA internal transcribed spacer region (ITS1-5.8S-ITS2 = ITS), revealed the occurrence of seven new phylogenetic species. These phylogenetic species also revealed unique combinations of phenotypes, allowing morphological distinction from their closest phylogenetic relatives. Four new species of Aquanectria and three new species of Gliocladiopsis are described and illustrated. Three of the four Aquanectria species deviate from the other species in the genus by having shorter conidia, which are in the size range observed in Gliocladiopsis species. They are placed in Aquanectria based on the phylogenetic analysis, but this also makes the morphological distinction between these two genera obsolete.


Assuntos
Hypocreales/classificação , Hypocreales/isolamento & purificação , Filogenia , Análise por Conglomerados , DNA Fúngico/química , DNA Fúngico/genética , DNA Ribossômico/química , DNA Ribossômico/genética , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , Equador , Microbiologia Ambiental , Guiana Francesa , Histonas/genética , Hypocreales/genética , Hypocreales/crescimento & desenvolvimento , Microscopia , Fator 1 de Elongação de Peptídeos/genética , RNA Ribossômico 5,8S/genética , Análise de Sequência de DNA , Singapura , Tubulina (Proteína)/genética
12.
Mycologia ; 111(2): 265-273, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30856060

RESUMO

A novel species of Microdochium was identified as the causal agent of a leaf blight of Poa pratensis (Kentucky blue grass) and Agrostis stolonifera (Creeping bentgrasses), two cold-season turfgrasses widely grown on golf courses in northern China. This disease first appears as small, water-soaked, and scattered leaf spots. Under conditions of high temperatures and successive days of rain, the infected leaves rapidly lose their integrity and large diseased patches appear. Fungal strains were isolated from blighted leaf spots. A phylogenetic analysis based on the nuc rDNA internal transcribed spacer regions and 5.8S rRNA gene (ITS1-5.8S-ITS2 = ITS) and parts of the ß-tubulin (TUB2) and RNA polymerase II second largest subunit (RPB2) genes strongly supported that these isolates are a distinct evolutionary lineage in Microdochium (Microdochiaceae, Xylariales) that represents a new taxonomic species, herein named as M. poae. Microscopic characters confirmed that these strains were morphologically distinct from known Microdochium species. The pathogenicity of M. poae was confirmed by inoculating spore suspension on both grasses and reisolation of the pathogen from symptomatic tissues. The optimal growth temperature suggests that the occurrence of the new leaf blight disease caused by M. poae was significantly different from the microdochium patch disease caused by M. nivale.


Assuntos
Agrostis/microbiologia , Doenças das Plantas/microbiologia , Poa/microbiologia , Xylariales/classificação , Xylariales/isolamento & purificação , China , Análise por Conglomerados , DNA Fúngico/química , DNA Fúngico/genética , DNA Ribossômico/química , DNA Ribossômico/genética , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , RNA Polimerases Dirigidas por DNA/genética , Kentucky , Microscopia , Filogenia , Folhas de Planta/microbiologia , RNA Ribossômico 5,8S/genética , Análise de Sequência de DNA , Tubulina (Proteína)/genética , Xylariales/citologia , Xylariales/genética
13.
J Parasitol ; 105(1): 180-185, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30817218

RESUMO

Pallisentis ( Brevitritospinus) indica Mittal and Lal, 1976 was found infecting the spotted snakehead Channa punctatus Bloch and Schneider (Channidae) from Meerut, Uttar Pradesh (U.P.), India. The species was identified on the basis of proboscis hooks, trunk spines, and other structures that separate it from all described species. Molecular analysis based on 18S rDNA placed the P. indica isolates within a clade of Pallisentis spp. but distinct from other representatives of the same genus. This study documents the first molecular characterization of P. indica from India.


Assuntos
Acantocéfalos/classificação , Acantocéfalos/genética , DNA de Helmintos/isolamento & purificação , Doenças dos Peixes/parasitologia , Helmintíase Animal/parasitologia , Acantocéfalos/anatomia & histologia , Animais , Sequência de Bases , Teorema de Bayes , Sequência Consenso , DNA de Helmintos/química , DNA Ribossômico/química , DNA Espaçador Ribossômico/química , Feminino , Peixes , Água Doce , Índia , Funções Verossimilhança , Masculino , Filogenia , RNA Ribossômico 18S/genética , RNA Ribossômico 5,8S/genética , Rios , Alinhamento de Sequência
14.
Mycologia ; 111(2): 319-338, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30908110

RESUMO

Seven species of bioluminescent fungi are recorded from the cloud forests in Mexico. Six represent new species belonging to the genus Mycena, whereas Mycena globulispora is a new distribution record for the country. Descriptions, illustrations, photographs, and an identification key to bioluminescent fungi species from Mexico are provided. Sequences of nuc rDNA internal transcribed spacer (ITS1-5.8S-ITS2 = ITS) region were generated for barcoding purposes and comparisons with similar species.


Assuntos
Agaricales/classificação , Agaricales/isolamento & purificação , Luminescência , Filogenia , Agaricales/genética , Agaricales/fisiologia , Análise por Conglomerados , DNA Fúngico/química , DNA Fúngico/genética , DNA Ribossômico/química , DNA Ribossômico/genética , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , México , RNA Ribossômico 5,8S/genética , Análise de Sequência de DNA
15.
Mycologia ; 111(2): 339-357, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30908115

RESUMO

Two new species, Fomitopsis mounceae and F. schrenkii (Polyporales, Basidiomycota) in the F. pinicola species complex in North America, are described and illustrated. Previous molecular phylogenetic analyses identified three well-delimited lineages that represent F. mounceae and F. ochracea from Canada, the Appalachian Mountains, and the northern United States and F. schrenkii from western and southwestern regions of the United States. Fomitopsis pinicola sensu stricto is restricted to Eurasia and does not occur in North America. Morphological descriptions of basidiocarps and cultures for F. mounceae, F. schrenkii, and F. ochracea are presented. The three species are readily differentiated by nuc rDNA internal transcribed spacer (ITS1-5.8S-ITS2 = ITS) sequence, geographic distribution, and basidiospore size. Polyporus ponderosus H. Schrenk is an earlier illegitimate synonym of F. schrenkii. Both F. mounceae and F. schrenkii have a heterothallic multiallelic incompatibility system.


Assuntos
Coriolaceae/classificação , Coriolaceae/isolamento & purificação , Canadá , Análise por Conglomerados , Coriolaceae/genética , Coriolaceae/crescimento & desenvolvimento , DNA Fúngico/química , DNA Fúngico/genética , DNA Ribossômico/química , DNA Ribossômico/genética , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , Carpóforos/crescimento & desenvolvimento , Genes Fúngicos , Filogeografia , Polyporus/classificação , RNA Ribossômico 5,8S/genética , Análise de Sequência de DNA , Estados Unidos
16.
Mycologia ; 111(2): 274-290, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30908119

RESUMO

In an investigation of the oomyceteous flora in rice paddy fields of Fars Province, Iran, three new Pythium species were isolated and identified on the basis of morphological features and molecular phylogenetic characteristics. Their unique morphological traits, including sexual and asexual structural characteristics (i.e., sporangial type; oogonial type and ornamentations; type and the number of antheridia per oogonium; and oospore type), cardinal temperatures, and colony morphology on various media, separated them from other known species. Using nuclear and mitochondrial genes, each species formed discrete lineages in phylogenetic analyses based on Bayesian inference and maximum likelihood methods. This paper describes these three new Pythium species, P. heteroogonium, P. longipapillum, and P. oryzicollum, and compares them with their related taxa via morphological features and molecular characteristics. Pathogenicity tests revealed the ability of P. oryzicollum to cause pre- and post-emergence damping-off, seed rot, crown rot, and reduced growth rate on rice.


Assuntos
Microscopia , Filogenia , Pythium/classificação , Pythium/isolamento & purificação , Microbiologia do Solo , Análise por Conglomerados , DNA Fúngico/química , DNA Fúngico/genética , DNA Ribossômico/química , DNA Ribossômico/genética , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , Complexo IV da Cadeia de Transporte de Elétrons/genética , Irã (Geográfico) , Técnicas Microbiológicas , Oryza/crescimento & desenvolvimento , Reação em Cadeia da Polimerase , Pythium/genética , Pythium/crescimento & desenvolvimento , RNA Ribossômico 5,8S/genética , Análise de Sequência de DNA , Tubulina (Proteína)/genética
17.
Acta Parasitol ; 64(1): 176-186, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30741381

RESUMO

BACKGROUND: Twelve species of Filisoma Van Cleave, 1928 are recognized parasitizing tropical and subtropical fish. Four of these species were described from kyphosid fish and it has been suggested that a co-speciation may have occurred among species of Kyphosus Lacepède, 1801 and Filisoma, which could provide valuable information about the evolution history of this host-parasite system. PURPOSE: During a survey of the helminth fauna of Kyphosus sectatrix (Linnaeus, 1758) and Kyphosus incisor (Cuvier, 1831) (Kyphosidae Jordan, 1887) off Rio de Janeiro coast, a new species of Filisoma was found and is described herein based on morphological, genetic, and ultrastructural data. METHODS: Fish were obtained off Rio de Janeiro coast, Brazil. The parasites found in the intestine were measured and drawings were made with a drawing tube. Type specimens were deposited at the Helminthological Collection of Oswaldo Cruz Institute (CHIOC). The ultrastructure was studied using scanning electron microscope. The genetic analysis included the study of the partial sequences of 18S, ITS1, 5.8S and 28S rDNA, and the mitochondrial cytochrome c oxidase 1 gene (cox 1), with phylogenetic reconstructions based on the maximum likelihood analysis. RESULTS: Filisoma caudata n. sp. is characterized by a proboscis with 16‒18 longitudinal rows of 38‒45 hooks each. Hooks are uniform in shape dorsoventrally, gradually decreasing in size towards the base of the proboscis. Anterior hooks are 30‒45 µ long, middle hooks 30‒35 µ long and 5 basal transversal hooks 20‒30 µ long. The new species is differentiated from the closest species Filisoma filiformis Weaver and Smales, 2013 by the size and distribution of hooks, apart from having a subterminal vulva and a curved posterior trunk end (tail) measuring 500‒1,000 long. Phylogenetic analysis based on 18S, 28S rDNA and mtDNA-cox1 markers grouped the new species with Filisoma bucerium Van Cleave, 1940 and Filisoma rizalinum Tubangui and Masiluñgan, 1946 showing a close relationship between these species of Cavisomidae Meyer, 1932 and Echinorhynchidae Cobbold, 1879; the latter represented by species of Acanthocephalus Koelreuther, 1771. The new species can be differentiated from others on morphological and molecular basis. A key to the 13 species of Filisoma Van Cleave, 1928 is provided. CONCLUSION: Filisoma caudata n. sp. is described herein based on morphological, genetic, and ultrastructural data. The topologies of obtained phylogenies suggest that species of Echinorhynchidae should be reevaluated since the family is considered paraphyletic in all analyses conducted.


Assuntos
Acantocéfalos/classificação , Acantocéfalos/isolamento & purificação , Doenças dos Peixes/parasitologia , Helmintíase Animal/parasitologia , Perciformes/parasitologia , Acantocéfalos/anatomia & histologia , Acantocéfalos/genética , Animais , Brasil , Análise por Conglomerados , DNA de Helmintos/química , DNA de Helmintos/genética , DNA Ribossômico/química , DNA Ribossômico/genética , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , Complexo IV da Cadeia de Transporte de Elétrons/genética , Estudos Longitudinais , Microscopia , Microscopia Eletrônica de Varredura , Filogenia , RNA Ribossômico 18S/genética , RNA Ribossômico 28S/genética , RNA Ribossômico 5,8S/genética , Análise de Sequência de DNA
18.
J Parasitol ; 105(1): 102-112, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30807727

RESUMO

Adults of the genus Austrodiplostomum are parasites in cormorants of the New World, whereas metacercariae are parasites from eye globe and brain of freshwater and brackish water fishes. In this study, specimens of Austrodiplostomum mordax from South America (type-species) were analyzed together with other specimens of Austrodiplostomum spp. collected from several locations across Middle America and North America. Partial DNA sequences of the mitochondrial gene cytochrome c oxidase subunit I ( COI), the internal transcribed spacers ( ITS1, ITS2, and 5.8S gene), and the D2-D3, domains of the large subunit ( LSU) of nuclear ribosomal DNA, were generated for both developmental stages and compared with available sequences of Austrodiplostomum spp. Phylogenetic analyses inferred with each molecular marker using maximum likelihood and Bayesian inference revealed the existence of 4 lineages representing 2 described species, A. mordax and Austrodiplostomum compactum (syn. Austrodiplostomum ostrowskiae) and 2 undescribed species of Austrodiplostomum recognized in previous studies. The COI haplotype network inferred with 172 sequences detected 28 haplotypes divided into 4 clusters, separating each other by 33 and 40 substitutions and with a genetic divergence ranging from 9 to 12%. The largest group included specimens identified as A. compactum plus those identified as A. ostrowskiae, supporting the synonymy of both species. As a result, we conclude that A. compactum is widely distributed across the Americas, in locations of the United States, Mexico, El Salvador, Honduras, Costa Rica, Venezuela, Peru, and Brazil. The other 2 undescribed species of the genus Austrodiplostomum were previously recorded in the United States and now are reported in Mexico. These 2 species cannot be described because adult forms have not been found in their definitive hosts. Finally, the species A. mordax has been found only in some lakes from Argentina, and it was validated in this study through molecular analyses.


Assuntos
Doenças das Aves/parasitologia , Complexo IV da Cadeia de Transporte de Elétrons/genética , Doenças dos Peixes/parasitologia , Genes Mitocondriais/genética , Trematódeos/classificação , Animais , Sequência de Bases , Teorema de Bayes , Aves , Encéfalo/parasitologia , América Central , DNA Intergênico/química , DNA Ribossômico/química , Peixes , Água Doce , Haplótipos , Funções Verossimilhança , Metacercárias/classificação , Metacercárias/genética , América do Norte , Filogenia , RNA Ribossômico 5,8S/genética , Águas Salinas , América do Sul , Trematódeos/genética , Corpo Vítreo/parasitologia
19.
Mycologia ; 111(1): 40-53, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30640586

RESUMO

Porodaedalea is a polypore genus of the Hymenochaetales that encompasses pathogens of conifer trees. In this study, we conduct a comprehensive study of the phylogeny and diversity of Porodaedalea based on collections and isolates from Europe, North America, North Africa, and Asia. Phylogenetic analysis of a two-gene data set, nuc ribosomal DNA internal transcribed spacers (ITS1-5.8S-ITS2 = ITS) and translation elongation factor 1-alpha (tef1), shows that 20 terminal clades that correspond to phylogenetic species well supported within Porodaedalea. Based on morphological evidence, five new species, P. alpicola, P. indica, P. kesiyae, P. microsperma, and P. yunnanensis, are described and illustrated. In addition, four still unnamed lineages are detected in North America and East Asia.


Assuntos
Basidiomycota/classificação , Variação Genética , Filogenia , África do Norte , Ásia , Basidiomycota/isolamento & purificação , Análise por Conglomerados , DNA Fúngico/genética , DNA Ribossômico/genética , Europa (Continente) , América do Norte , Filogeografia , RNA Ribossômico 5,8S/genética , Análise de Sequência de DNA , Traqueófitas/microbiologia
20.
Mol Ecol Resour ; 19(1): 118-127, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30240145

RESUMO

Sequence comparison and analysis of the various ribosomal genetic markers are the dominant molecular methods for identification and description of fungi. However, new environmental fungal lineages known only from DNA data reveal significant gaps in our sampling of the fungal kingdom in terms of both taxonomy and marker coverage in the reference sequence databases. To facilitate the integration of reference data from all of the ribosomal markers, we present three sets of general primers that allow for amplification of the complete ribosomal operon from the ribosomal tandem repeats. The primers cover all ribosomal markers: ETS, SSU, ITS1, 5.8S, ITS2, LSU and IGS. We coupled these primers successfully with third-generation sequencing (PacBio and Nanopore sequencing) to showcase our approach on authentic fungal herbarium specimens (Basidiomycota), aquatic chytrids (Chytridiomycota) and a poorly understood lineage of early diverging fungi (Nephridiophagidae). In particular, we were able to generate high-quality reference data with Nanopore sequencing in a high-throughput manner, showing that the generation of reference data can be achieved on a regular desktop computer without the involvement of any large-scale sequencing facility. The quality of the Nanopore generated sequences was 99.85%, which is comparable with the 99.78% accuracy described for Sanger sequencing. With this work, we hope to stimulate the generation of a new comprehensive standard of ribosomal reference data with the ultimate aim to close the huge gaps in our reference datasets.


Assuntos
Código de Barras de DNA Taxonômico/métodos , DNA Ribossômico/genética , Fungos/classificação , Fungos/genética , Genes de RNAr , Sequências de Repetição em Tandem , Análise por Conglomerados , Primers do DNA/genética , DNA Fúngico/química , DNA Fúngico/genética , DNA Ribossômico/química , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , Sequenciamento de Nucleotídeos em Larga Escala , Filogenia , RNA Ribossômico/genética , RNA Ribossômico 18S/genética , RNA Ribossômico 5,8S/genética , Análise de Sequência de DNA
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