Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 2.547
Filtrar
1.
Epidemiol Infect ; 148: e49, 2020 02 14.
Artigo em Inglês | MEDLINE | ID: mdl-32054545

RESUMO

A new fast-growing mycobacterium, designated strain QGD101T, was isolated from the sputum of an 84-year-old man suspected of tuberculosis in Wuhan Medical Treatment Center, Hubei, China. This strain was a gram-staining-negative, aerobic, non-spore-forming and catalase-positive bacterium, which was further identified as the NTM by PNB and TCH tests. The moxifloxacin and levofloxacin exhibited strong suppressing function against QGD101T with MIC values of 0.06 and 0.125 µg/ml after drug susceptibility testing of six main antimicrobial agents on mycobacteria. Based on the sequence analysis of 16S rRNA, rpoB, hsp65 and 16S-23S rRNA internal transcribed spacer, the strain QGD101T could not be identified to a species level. Mycobacterium moriokaense ATCC43059T that shared the highest 16S rRNA gene sequence similarity (98%) with strain QGD101T was actually different in genomes average nucleotide identity (78.74%). In addition, the major cellular fatty acids of QGD101T were determined as C18:1ω9c, C16:0 and C18:2ω6c. The DNA G + C content was 64.9% measured by high performance liquid chromatography. Therefore, the phenotypic and genotypic characterisation of this strain led us to the conclusion that it represents a novel species of mycobacteria, for which the name Mycobacterium hubeiense sp. nov. (type strain QGD101T = CCTCCAA 2017003T = KCTC39927T) was proposed. Thus, the results of this study are very significant for the clinical diagnosis of tuberculosis and future personalised medicine.


Assuntos
Infecções por Mycobacterium não Tuberculosas/microbiologia , Mycobacterium/classificação , Mycobacterium/isolamento & purificação , Escarro/microbiologia , Idoso de 80 Anos ou mais , Antituberculosos/farmacologia , Proteínas de Bactérias/genética , Composição de Bases , Chaperonina 60/genética , China , Análise por Conglomerados , Citosol/química , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , RNA Polimerases Dirigidas por DNA/genética , Ácidos Graxos/análise , Humanos , Levofloxacino/farmacologia , Masculino , Testes de Sensibilidade Microbiana , Moxifloxacina/farmacologia , Mycobacterium/genética , Mycobacterium/fisiologia , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
2.
Exp Parasitol ; 209: 107814, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31816280

RESUMO

Giardia duodenalis is one of the most important intestinal parasites globally, especially in children, and in Cuba is the leading cause of chronic paediatric diarrhoea in this population. G. duodenalis is composed of eight genetic groups (or assemblages), two of which (A and B) are apparently zoonotic, occurring in both humans and other animals. However, consensus on the most appropriate genotyping scheme for optimal characterization of G. duodenalis isolates is lacking. In this article we present the results of three descriptive observational studies conducted in Havana, Cuba between 2010 and 2013, with the aim of comparing the results from molecular (PCR) approaches targeting different genes in order to assign with confidence 224 isolates of G. duodenalis to the correct assemblages. In each sub-study, following DNA isolation by the phenol/chloroform/isoamyl alcohol extraction method, PCR targeting the triose phosphate isomerase (tpi) gene was used for molecular characterization, as well as one additional PCR-method targeting another gene or pair of genes. DNA amplification was obtained in 87%, 83%, and 80% in the three sub-studies. Although excellent agreement (kappa index = 1) was recorded between results from some pairs of genes, for other combinations only moderate or substantial agreement was achieved. These results highlight the importance of interpretation of genotyping data, especially when single genetic markers are used. From the results of our studies, PCR targeting a combination of the tpi gene and the intergenic spacer region of rDNA may be a useful approach for the molecular characterization of G. duodenalis isolates.


Assuntos
Técnicas de Genotipagem/normas , Giardia lamblia/classificação , Giardíase/parasitologia , Reação em Cadeia da Polimerase/métodos , Animais , Criança , Pré-Escolar , Cuba , DNA de Protozoário/química , DNA de Protozoário/isolamento & purificação , DNA Espaçador Ribossômico/química , Fezes/parasitologia , Giardia lamblia/genética , Giardia lamblia/isolamento & purificação , Glutamato Desidrogenase/genética , Humanos , Polimorfismo de Fragmento de Restrição , Triose-Fosfato Isomerase/genética
3.
J Parasitol ; 105(6): 882-889, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31738125

RESUMO

The genus Eustrongylides includes nematodes known as the etiological agent of the "big red worm disease." The aim of this work was to identify Eustrongylides spp. larvae from fish and adults from great cormorant (Phalacrocorax carbo) sampled at Lake Trasimeno, Italy, by morphological and molecular analysis. Histopathological description of the lesions in birds was also provided. We described adults of Eustrongylides excisus for the first time in Italy, and we also linked larval stages 3 and 4 to adults. The use of molecular tools combined with the traditional taxonomy will help the identification of the species, including species inquirendae. Moreover, molecular analysis can also help to investigate the role of intermediate and paratenic hosts, to deepen the knowledge about geographical distribution of the different Eustrongylides spp. and to define the zoonotic potential of E. excisus, which has not yet been identified as causal agent of human cases.


Assuntos
Doenças das Aves/parasitologia , Dioctophymatoidea/anatomia & histologia , Dioctophymatoidea/genética , Infecções por Enoplida/veterinária , Doenças dos Peixes/parasitologia , Animais , Doenças das Aves/patologia , Aves , DNA Espaçador Ribossômico/química , Dioctophymatoidea/classificação , Dioctophymatoidea/crescimento & desenvolvimento , Infecções por Enoplida/parasitologia , Infecções por Enoplida/patologia , Feminino , Doenças dos Peixes/patologia , Peixes , Água Doce , Itália , Lagos , Larva/anatomia & histologia , Larva/classificação , Larva/genética , Funções Verossimilhança , Masculino , Óvulo/ultraestrutura , Reação em Cadeia da Polimerase , Proventrículo/parasitologia , Proventrículo/patologia
4.
J Parasitol ; 105(5): 769-782, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31625813

RESUMO

Truttaedacnitis truttae is a cucullanid nematode of primarily salmonine fishes. Brown trout (Salmo trutta) in Europe reportedly become parasitized by ingesting lampreys (Lampetra planeri) carrying infective larvae. However, our field and laboratory observations suggested that North American specimens of T. truttae have an alternative life cycle. High abundances and potential impact of T. truttae in rainbow trout, Oncorhynchus mykiss, in the Colorado River drainage in Grand Canyon, where there are no lampreys, prompted a study on the transmission dynamics of this nematode. Eggs of T. truttae, collected from live gravid females, were incubated in the laboratory. Snails, Physa gyrina and Lymnaea sp., were exposed to T. truttae larvae 3-4 wk later. Active larvae of T. truttae were observed penetrating the intestinal wall of exposed snails, and worm larvae were found in the visceral tissues when examined 1 wk after exposure. Larvae in snails showed little growth and development 2 wk later and corresponded to L3 larvae. Infected snails were fed to hatchery-reared juvenile rainbow trout. Developing stages were subsequently found in the mucosal lining and lumen of trout intestines. Adult male and female (gravid) worms were found in the ceca of trout examined 5-6 mo after consuming infected snails. Larvae found in pepsin/trypsin digests and mucosal scrapings from wild, naturally infected, trout corroborate laboratory findings. Screening of Physa sp. and gammarids collected from Colorado River, Grand Canyon, for natural infections with T. truttae using the ITS1 rDNA marker gave positive results. Truttaedacnitis truttae is the second species, after Truttaedacnitis clitellarius of lake sturgeon, capable of using a snail first intermediate/paratenic host and is similar to several other cucullanids in having a histotropic phase of development in the definitive fish host.


Assuntos
Doenças dos Peixes/parasitologia , Estágios do Ciclo de Vida , Caramujos/parasitologia , Infecções por Spirurida/veterinária , Spirurina/crescimento & desenvolvimento , Truta/parasitologia , Animais , Ceco/parasitologia , DNA de Helmintos/isolamento & purificação , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/isolamento & purificação , Feminino , Larva/crescimento & desenvolvimento , Masculino , Oncorhynchus mykiss/parasitologia , Projetos Piloto , Reação em Cadeia da Polimerase/veterinária , Rios , Infecções por Spirurida/parasitologia , Spirurina/anatomia & histologia
5.
Int J Parasitol ; 49(12): 945-961, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31628939

RESUMO

We explore the growing issue of cryptic speciation in the Digenea through study of museum material and newly collected specimens consistent with the enenterid genus Koseiria from five species of the Kyphosidae and Chaetodontoplus meredithi Kuiter (Pomacanthidae) collected in the Indo-Pacific. We use an integrated approach, employing traditional morphometrics, principal components analysis (PCA), and molecular data (ITS2 and 28S rDNA). Our results support recombination of Koseiria allanwilliamsi Bray & Cribb, 2002 as Proenenterum allanwilliamsi (Bray & Cribb, 2002) n. comb. and transfer of Koseiria huxleyi Bray & Cribb, 2001 to a new genus as Enenterageitus huxleyi (Bray & Cribb, 2002) n. comb. Molecular data indicate the presence of four further species consistent with Koseiria, one from Western Australia (sequence data only) and three from eastern Australia. All three eastern Australian species are morphologically consistent with Koseiria xishaensis Gu & Shen, 1983, but distinct from all other previously described species. Although K. xishaensis has been reported from Australia, we conclude that the similarity of the present forms to the original description of K. xishaensis means records of this species from Japan, Palau and Australia are unreliable. Because the eastern Australian forms cannot be reliably ascribed to K. xishaensis, we describe Koseiria argalea n. sp., Koseiria laiphopharophora n. sp., and Koseiria pyknophora n. sp., following application of PCAs and iterative refinement of species concepts and type series. These analyses did not allow convincing identification hypotheses for all specimens examined. In this genus, both morphological and molecular data, together with reliable host identifications, are essential for species recognition, and thus we refrain from attempting to name samples lacking molecular data. The issues presented by these taxa encapsulate those of trematodes in the region as a whole. Many records require dramatically improved supporting data, leading to substantial uncertainly in the identification of this fauna.


Assuntos
Peixes/parasitologia , Filogenia , Trematódeos/anatomia & histologia , Trematódeos/classificação , Animais , Biometria , 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 , Microscopia , Oceano Pacífico , RNA Ribossômico 28S/genética , Análise de Sequência de DNA , Trematódeos/genética , Trematódeos/isolamento & purificação
6.
Exp Parasitol ; 206: 107771, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31585116

RESUMO

A PCR targeting mitochondrial cytochrome oxidase subunit III (cox3) for molecular detection of Babesia gibsoni infection in dogs has been developed in this study. Fifty blood samples from suspected clinical cases from dogs, brought to the veterinary college clinics, were examined for presence of B. gibsoni using conventional diagnosis by microscopic examination of Giemsa stained thin blood smears. In addition, species specific PCRs targeting ITS-1 region (BgITS-1 PCR) and nested PCR targeting 18S ribosomal RNA gene (Bg18SnPCR) were carried out. A 634 bp PCR fragment of B. gibsoni cox3 gene was amplified in positive samples from three geographical locations of Satara, Wai and Pune in Maharashtra state of India. From analysis of the sequence of the B. gibsoni cox3 gene, we found that the Indian isolate had 96-98% similarity to the isolate from Japan and China. Post sequencing, de-novo diagnostic primer pair for species specific amplification of 164 bp fragment of B. gibsonicox3 was designed and the PCR was standardized. The diagnostic results of de-novo Bgcox3 PCR were compared with BgITS-1 PCR and Bg18S nPCR. Thin blood smears detected 22% (11/50) samples positive for small form of Babesia species. The BgITS-1 PCR detected 25% samples (15/50) as positive and Bg18S nPCR detected 80% (40/50) B. gibsoni positive samples. The de-novo Bgcox3 PCR detected 66% (33/50) samples positive for B. gibsoni (at 95% CI). The analytical sensitivity of cox3 PCR was evaluated as 0.000003% parasitaemia or 09 parasites in 100  µl of blood. The de-novo diagnostic cox3 PCR did not cross react with control positive DNA from other haemoprotozoa and rickettsia like B. vogeli, Hepatozoon canis, Trypanosoma evansi, Ehrlichia canis and Anaplasma platys. Statistically, cox3 PCR had better diagnostic efficiency than ITS-1 PCR in terms of sensitivity (p = 0.0006). No statistically significant difference between results of cox3 PCR and 18S nPCR was observed (p = 0.1760). Kappa values estimated for each test pair showed fair to moderate agreement between the observations. Specificity of Bgcox3 PCR was 100% when compared with microscopy or BgITS-1 PCR. Sensitivity of Bgcox3 PCR was 100% when compared with that of Bg18S nPCR.


Assuntos
Babesia/isolamento & purificação , Babesiose/diagnóstico , Doenças do Cão/diagnóstico , Complexo IV da Cadeia de Transporte de Elétrons/genética , Mitocôndrias/enzimologia , Animais , Babesia/classificação , Babesia/genética , Babesiose/parasitologia , Sequência de Bases , Reações Cruzadas , DNA Espaçador Ribossômico/química , Doenças do Cão/parasitologia , Cães , Eritrócitos/parasitologia , Funções Verossimilhança , Filogenia , Reação em Cadeia da Polimerase/veterinária , Valor Preditivo dos Testes , RNA Ribossômico 18S/análise , Sensibilidade e Especificidade , Alinhamento de Sequência/veterinária
7.
mBio ; 10(5)2019 10 08.
Artigo em Inglês | MEDLINE | ID: mdl-31594815

RESUMO

The wild legume Lotus japonicus engages in mutualistic symbiotic relationships with arbuscular mycorrhiza (AM) fungi and nitrogen-fixing rhizobia. Using plants grown in natural soil and community profiling of bacterial 16S rRNA genes and fungal internal transcribed spacers (ITSs), we examined the role of the Lotus symbiosis genes RAM1, NFR5, SYMRK, and CCaMK in structuring bacterial and fungal root-associated communities. We found host genotype-dependent community shifts in the root and rhizosphere compartments that were mainly confined to bacteria in nfr5 or fungi in ram1 mutants, while symrk and ccamk plants displayed major changes across both microbial kingdoms. We observed in all AM mutant roots an almost complete depletion of a large number of Glomeromycota taxa that was accompanied by a concomitant enrichment of Helotiales and Nectriaceae fungi, suggesting compensatory niche replacement within the fungal community. A subset of Glomeromycota whose colonization is strictly dependent on the common symbiosis pathway was retained in ram1 mutants, indicating that RAM1 is dispensable for intraradical colonization by some Glomeromycota fungi. However, intraradical colonization by bacteria belonging to the Burkholderiaceae and Anaeroplasmataceae is dependent on AM root infection, revealing a microbial interkingdom interaction. Despite the overall robustness of the bacterial root microbiota against major changes in the composition of root-associated fungal assemblages, bacterial and fungal cooccurrence network analysis demonstrates that simultaneous disruption of AM and rhizobium symbiosis increases the connectivity among taxa of the bacterial root microbiota. Our findings imply a broad role for Lotus symbiosis genes in structuring the root microbiota and identify unexpected microbial interkingdom interactions between root symbionts and commensal communities.IMPORTANCE Studies on symbiosis genes in plants typically focus on binary interactions between roots and soilborne nitrogen-fixing rhizobia or mycorrhizal fungi in laboratory environments. We utilized wild type and symbiosis mutants of a model legume, grown in natural soil, in which bacterial, fungal, or both symbioses are impaired to examine potential interactions between the symbionts and commensal microorganisms of the root microbiota when grown in natural soil. This revealed microbial interkingdom interactions between the root symbionts and fungal as well as bacterial commensal communities. Nevertheless, the bacterial root microbiota remains largely robust when fungal symbiosis is impaired. Our work implies a broad role for host symbiosis genes in structuring the root microbiota of legumes.


Assuntos
Bactérias/crescimento & desenvolvimento , Fungos/crescimento & desenvolvimento , Interações entre Hospedeiro e Microrganismos , Lotus/microbiologia , Interações Microbianas , Proteínas de Plantas/genética , Simbiose , Bactérias/classificação , Bactérias/genética , Análise por Conglomerados , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Fúngico/química , DNA Fúngico/genética , DNA de Plantas/química , DNA de Plantas/genética , DNA Ribossômico/química , DNA Ribossômico/genética , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , Fungos/classificação , Fungos/genética , Genótipo , Lotus/genética , Filogenia , Raízes de Plantas/microbiologia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Microbiologia do Solo
8.
Genet Sel Evol ; 51(1): 59, 2019 Oct 26.
Artigo em Inglês | MEDLINE | ID: mdl-31655542

RESUMO

BACKGROUND: Ribosomal DNA (rDNA) repeats are situated in the nucleolus organizer regions (NOR) of chromosomes and transcribed into rRNA for ribosome biogenesis. Thus, they are an essential component of eukaryotic genomes. rDNA repeat units consist of rRNA gene clusters that are transcribed into single pre-rRNA molecules, each separated by intergenic spacers (IGS) that contain regulatory elements for rRNA gene cluster transcription. Because of their high repeat content, rDNA sequences are usually absent from genome assemblies. In this work, we used the long-read sequencing technology to describe the chicken IGS and fill the knowledge gap on rDNA sequences of one of the key domesticated animals. METHODS: We used the long-read PacBio RSII technique to sequence the BAC clone WAG137G04 (Wageningen BAC library) known to contain chicken NOR elements and the HGAP workflow software suit to assemble the PacBio RSII reads. Whole-genome sequence contigs homologous to the chicken rDNA repetitive unit were identified based on the Gallus_gallus-5.0 assembly with BLAST. We used the Geneious 9.0.5 and Mega software, maximum likelihood method and Chickspress project for sequence evolution analysis, phylogenetic tree construction and analysis of the raw transcriptome data. RESULTS: Three complete IGS sequences in the White Leghorn chicken genome and one IGS sequence in the red junglefowl contig AADN04001305.1 (Gallus_gallus-5.0) were detected. They had various lengths and contained three groups of tandem repeats (some of them being very GC rich) that form highly organized arrays. Initiation and termination sites of rDNA transcription were located within small and large unique regions (SUR and LUR), respectively. No functionally significant sites were detected within the tandem repeat sequences. CONCLUSIONS: Due to the highly organized GC-rich repeats, the structure of the chicken IGS differs from that of IGS in human, apes, Xenopus or fish rDNA. However, the chicken IGS shares some molecular organization features with that of the turtles, which are other representatives of the Sauropsida clade that includes birds and reptiles. Our current results on the structure of chicken IGS together with the previously reported ribosomal gene cluster sequence provide sufficient data to consider that the complete chicken rDNA sequence is assembled with confidence in terms of molecular DNA organization.


Assuntos
Galinhas/genética , DNA Espaçador Ribossômico/genética , DNA Ribossômico/genética , Animais , Sequência Conservada , DNA Espaçador Ribossômico/química , Homologia de Sequência do Ácido Nucleico
9.
Vet Parasitol ; 275: 108933, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31606485

RESUMO

We have validated ITS-2 rDNA nemabiome next-generation amplicon sequencing to determine relative species abundance of gastrointestinal nematode species in ovine fecal samples. In order to determine species representation biases, ITS-2 rDNA amplicon sequencing was applied to mock communities or field populations with known proportions of L3 for eight of the major ovine gastrointestinal nematode species: Teladorsagia circumcincta, Trichostrongylus vitrinus, Haemonchus contortus, Cooperia curticei, Trichostrongylus axei, Trichostrongylus colubriformis, Chabertia ovina and Oesophagostumum venulosum. Correction factors, calculated from this data, were shown to reduce species representation biases when applied to an independent set of field samples of known composition. We compared ITS-2 rDNA amplicon sequencing data that was generated from harvested eggs, freshly hatched L1 or L3 larvae following fecal culture and no statistically significant differences were found for the more abundant parasite species. We then applied the validated ITS-2 rDNA nemabiome amplicon sequencing assay to a set of archived L1 gastrointestinal nematode populations, collected in 2008 from fecal samples from 93 groups of 20 ewes and 61 groups of 20 lambs derived from 99 UK sheep farms. The presence of the major gastrointestinal nematode species had previously been determined on this large sample set by species-specific PCR. We show how the ITS-2rDNA amplicon sequencing data provided much more detailed information on species abundance than the previous species-specific PCR. This new data represents the most comprehensive overview of the relative abundance of the major gastrointestinal nematode species across UK sheep farms to date. Substantial variance in the relative abundance of both T. circumcincta and T. vitrinus between farms was revealed with the former species being of statistically significantly higher abundance in all three regions sampled (England, Scotland and Wales). The data also revealed that the relative abundance of T. circumcinta was statistically significantly higher in ewes than in lambs with the opposite pattern being the case for T. vitrinus. The nemabiome sequencing data also clearly illustrated the sporadic nature and skewed distribution of H. controtus across UK sheep farms as well as a higher relative abundance on farms from England compared to Wales and Scotland. The nemabiome survey also provides the first widescale data on the relative abundance of the two major large intestinal nematodes C. ovina and O. venulosum. This work validates ITS-2 rDNA nemabiome sequencing for use in sheep and illustrates the power of the approach for large scale surveillance of ovine gastrointestinal nematodes.


Assuntos
DNA Espaçador Ribossômico/química , Microbioma Gastrointestinal/genética , Enteropatias Parasitárias/veterinária , Infecções por Nematoides/veterinária , Doenças dos Ovinos/parasitologia , Abomaso/parasitologia , Animais , Biologia Computacional , DNA Espaçador Ribossômico/genética , Feminino , Enteropatias Parasitárias/epidemiologia , Enteropatias Parasitárias/parasitologia , Intestino Grosso/parasitologia , Intestino Delgado/parasitologia , Larva/genética , Infecções por Nematoides/epidemiologia , Infecções por Nematoides/parasitologia , Óvulo , Reação em Cadeia da Polimerase/veterinária , Prevalência , Ovinos , Doenças dos Ovinos/epidemiologia , Especificidade da Espécie , Reino Unido/epidemiologia
10.
Mycologia ; 111(5): 813-831, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31525126

RESUMO

The genus Antrodiella includes resupinate and pileate species of polypores with a dimitic hyphal system, small, globose to cylindrical basidiospores, absence of cystidia, tetrapolar mating system, and haplo-dikaryotic nuclear behavior. Recent studies, however, indicate that Antrodiella is highly polyphyletic, so many of its species have been transferred to other genera. This study reviews the systematic status and diversity of Antrodiella from the Neotropics based, in part, on studies of type specimens. Collections from Brazil were used for molecular analysis of nuc rDNA internal transcribed spacer region ITS1-5.8S-ITS2 (ITS), nuc 28S rDNA (28S), and portions of genes encoding translation elongation factor 1-α (tef1) and the second largest subunit of RNA polymerase II (rpb2). Eight genera are confirmed to include Neotropical species treated as Antrodiella in a broad sense: Aegis, Antrodiella s. str., Flaviporus, Metuloidea, Mycorrhaphium, Rickiopora, Trametopsis, and Trullella. Molecular data reveal the occurrence of two new species, described as Antrodiella trivialis, the only Neotropical species of Antrodiella s. str. known so far, and Mycorrhaphium hispidum. In addition, Antrodiella luteocontexta was found to nest in the genus Aegis, close to the Grifolaceae and Polyporaceae; therefore, the new combination Aegis luteocontexta is proposed. Comments on the eight Antrodiella-related genera as well as species with uncertain taxonomic position are provided, together with a key to their identification.


Assuntos
Variação Genética , Filogenia , Polyporales/classificação , Polyporales/genética , Brasil , Análise por Conglomerados , DNA Fúngico/química , DNA Fúngico/genética , DNA Ribossômico/química , DNA Ribossômico/genética , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , Fator 1 de Elongação de Peptídeos/genética , Polyporales/isolamento & purificação , RNA Polimerase II/genética , RNA Ribossômico 28S/genética , Análise de Sequência de DNA , Clima Tropical
11.
Mycologia ; 111(5): 782-792, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31545143

RESUMO

We present an account of Rhizopogon introduced from plantings of exotic pine plantations in Argentine Patagonia. Nuc rDNA internal transcribed spacer ITS1-5.8S-ITS2 (ITS) and nuc 28S rDNA (28S) sequences were used to identify specimens from Argentina and examine their relationships with geographically different Rhizopogon species. Based on phylogenetic analyses, we confirm that four species of Rhizopogon occur in pine plantations across Patagonia. Several Rhizopogon collections from Pinus ponderosa plantations across different provinces cluster with R. arctostaphyli, a species within R. subg. Amylopogon. The majority of Patagonian Rhizopogon, however, form three different lineages in R. subg. Roseoli. The first of these, R. roseolus sensu Trappe, includes numerous collections from Pinus ponderosa, P. contorta, and P. radiata stands of North American affiliation. The second, R. roseolus sensu Martin and Garcia from P. ponderosa plantations, clusters in clade IIIa of the R. roseolus complex, which also includes the holotype collection of R. mohelnensis from the Czech Republic. The third species in R. subg. Roseoli, and fourth species overall from Patagonia, is R. granuloflavus from Pinus ponderosa plantations. Multiplex polymerase chain reaction (PCR) of numerous Roseoli samples failed to produce an amplicon indicative of either Japanese or New Zealand shoro.


Assuntos
Basidiomycota/classificação , Basidiomycota/isolamento & purificação , Variação Genética , Filogenia , Pinus ponderosa/microbiologia , Argentina , Basidiomycota/genética , Análise por Conglomerados , DNA Fúngico/química , DNA Fúngico/genética , DNA Ribossômico/química , DNA Ribossômico/genética , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , RNA Ribossômico 28S/genética , Análise de Sequência de DNA
12.
Korean J Parasitol ; 57(4): 417-422, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31533409

RESUMO

From October 2015 to August 2018, tapeworm proglottids were obtained from 10 patients who were residents of Daegu and Gyeongbuk provinces and had a history of raw beef consumption. Most of them had no overseas travel experience. The gravid proglottids obtained from the 10 cases had 15-20 lateral uterine branches. A part of internal transcribed spacer 1 (ITS1) DNA of the 10 cases, amplified by polymerase chain reaction (PCR) and digested with AleI restriction enzyme, produced the same band pattern of Taenia saginata, which differentiated from T. asiatica and T. solium. Sequences of ITS1 and cytochrome c oxidase subunit 1 (cox1) showed higher homology to T. saginata than to T. asiatica and T. solium. Collectively, these 10 cases were identified as T. saginata human infections. As taeniasis is one of the important parasitic diseases in humans, it is necessary to maintain hygienic conditions during livestock farming to avoid public health concerns.


Assuntos
DNA Espaçador Ribossômico/análise , Taenia saginata/isolamento & purificação , Teníase/diagnóstico , Adulto , Idoso , Animais , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , Complexo IV da Cadeia de Transporte de Elétrons/química , Complexo IV da Cadeia de Transporte de Elétrons/genética , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , República da Coreia , Mapeamento por Restrição , Homologia de Sequência , Taenia saginata/classificação , Taenia saginata/genética , Teníase/parasitologia , Adulto Jovem
13.
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
14.
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
15.
Mycologia ; 111(5): 871-883, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31433734

RESUMO

Most known brown rot-producing species of Polyporales belong to the so-called "Antrodia clade" that largely consists of poroid species. In this study, we use three genetic markers to revise Antrodia s. str., the core group of this clade. We show that a corticioid species with a smooth hymenophore, Phlebia griseoflavescens, belongs to Antrodia s. str. Accordingly, we revise the generic concept of Antrodia s. str. to accommodate this species and two recently described poroid taxa, A. tenerifensis and A. multiformis. In addition, we describe two new poroid species within Antrodia s. str., A. latebrosa from Africa and A. peregrina from East Asia, and provide new documentation for the Southeast Asian species A. parvula based on recent collections from the type location.


Assuntos
Antrodia/classificação , Antrodia/genética , Antrodia/citologia , Antrodia/isolamento & purificação , Análise por Conglomerados , DNA Fúngico/química , DNA Fúngico/genética , DNA Ribossômico/química , DNA Ribossômico/genética , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , Microbiologia Ambiental , Microscopia , Filogenia , Análise de Sequência de DNA
16.
Mycopathologia ; 184(5): 701-706, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31376041

RESUMO

Mycelial basidiomycetes rarely produce mycoses in animals including humans. We report a case of a 9-year-old female mongrel dog with lesions in the prescapular lymph nodes. The histopathology of a lymph node sample showed flexuous septate hyphae, and a sterile mold grew in culture from that specimen. DNA sequencing of the ITS region allowed us to identify the fungus as Tropicoporus tropicalis. The dog was treated with itraconazole, but it was euthanized six months later due to an unfavorable clinical outcome. Tropicoporus tropicalis is an infrequent pathogen of pets, and the use of molecular tools is needed for its identification. Animal infections due to T. tropicalis were not previously been reported in Argentina.


Assuntos
Basidiomycota/isolamento & purificação , Doenças do Cão/diagnóstico , Doenças do Cão/patologia , Micoses/patologia , Micoses/veterinária , Animais , Antifúngicos/administração & dosagem , Argentina , Basidiomycota/classificação , Basidiomycota/genética , DNA Fúngico/química , DNA Fúngico/genética , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , Doenças do Cão/tratamento farmacológico , Cães , Feminino , Histocitoquímica , Itraconazol/administração & dosagem , Linfonodos/patologia , Técnicas Microbiológicas , Micoses/tratamento farmacológico , Análise de Sequência de DNA , Falha de Tratamento
17.
Mycologia ; 111(5): 730-747, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31449474

RESUMO

Ganoderma is a cosmopolitan genus that encompasses species with cultural, economic, and pathogenic importance. Despite the importance of this genus, knowledge pertaining to the species diversity of Ganoderma in South Africa is limited. This study aimed at elucidating the identity and phylogenetic placements of Ganoderma samples obtained during a survey of wood-rotting fungi in the Garden Route National Park (GRNP) of South Africa, supplemented with isolates obtained from other localities across the country. Identification was achieved by means of multilocus phylogenetic inference combined with morphological evaluation. In total, eight distinct species of Ganoderma were recovered from different hosts and localities across the country. Of these, Ganoderma cf. cupreum and Ganoderma cf. resinaceum represent possible new records for South Africa. Two novel species are described, namely, G. eickeri. and G. knysnamense. Ganoderma eickeri, sp. nov., is characterized by a triquetrous and broadly attached basidiome, a sulcate or zonate yellowish brown to brown pilear surface, and ovoid to ellipsoid basidiospores. Ganoderma knysnamense is distinguished by an applanate to ungulate, sometimes convex, dimidiate to broadly attached basidiome, a chocolate-brown pilear surface covered with a hard woody-like crust and ellipsoid, broadly ellipsoid to ovoid basidiospores. The discovery of two new Ganoderma species in this study raises the known Ganoderma species in South Africa to 13.


Assuntos
Ganoderma/classificação , Ganoderma/isolamento & purificação , Micobioma , Filogenia , Madeira/microbiologia , Análise por Conglomerados , DNA Fúngico/química , DNA Fúngico/genética , DNA Ribossômico/química , DNA Ribossômico/genética , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , Ganoderma/genética , Ganoderma/crescimento & desenvolvimento , Microscopia , Parques Recreativos , Fator 1 de Elongação de Peptídeos/genética , Análise de Sequência de DNA , África do Sul , Esporos Fúngicos/citologia , Tubulina (Proteína)/genética
18.
Mycologia ; 111(5): 832-856, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31460851

RESUMO

Two new species and a new combination of Hypoxylon from Texas were identified and described based on morphological, multigene phylogenetic (ITS [nuc rDNA internal transcribed spacer region ITS1-5.8S-ITS2], 28S [5' 1200 bp of nuc 28S rDNA], RPB2 [partial second largest subunit of the DNA-directed RNA polymerase II], TUB2 [partial ß-tubulin]), and chemotaxonomic data. Hypoxylon olivaceopigmentum is characterized by its pulvinate to glomerate stromata, olivaceous KOH-extractable pigments, equilateral ascospores, and indehiscent perispore. Hypoxylon texense can be distinguished from morphologically similar species by its rust to dark brick KOH-extractable pigments and the high-performance liquid chromatography (HPLC) profile of its stromatal secondary metabolites. Hypoxylon hinnuleum is proposed as the sexual morph of Nodulisporium hinnuleum, featuring dark vinaceous glomerate stromata with dark brick KOH-extractable pigments composed of cohaerin-type azaphilones and smooth equilateral ascospores with indehiscent perispore. Based on these diagnostic characters, H. hinnuleum forms a complex with H. croceum and H. minicroceum. More than 50 ITS sequences with high identity originating from North American and East Asian environmental isolates formed a well-supported clade with the type of N. hinnuleum, demonstrating the widespread distribution of the species complex. In addition, updated descriptions and comprehensive illustrations with detailed information on the diagnostic features of H. fendleri and H. perforatum are provided. The multilocus phylogenetic reconstruction of Hypoxylon supported the status of the new species and broadened the knowledge about intergeneric relationships.


Assuntos
Microbiologia Ambiental , Filogenia , Esporos Fúngicos/citologia , Xylariales/classificação , Xylariales/isolamento & purificação , Análise por Conglomerados , DNA Fúngico/química , DNA Fúngico/genética , DNA Ribossômico/química , DNA Ribossômico/genética , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , Microscopia , Pigmentos Biológicos/análise , RNA Polimerase II/genética , RNA Ribossômico 28S/genética , Análise de Sequência de DNA , Texas , Tubulina (Proteína)/genética , Xylariales/genética , Xylariales/fisiologia
19.
Histochem Cell Biol ; 152(4): 271-280, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31346697

RESUMO

In human cells, the intergenic spacers (IGS), which separate ribosomal genes, are complex approximately 30 kb-long loci. Recent studies indicate that all, or almost all, parts of IGS may be transcribed, and that at least some of them are involved in the regulation of the ribosomal DNA (rDNA) transcription, maintenance of the nucleolar architecture, and response of the cell nucleus to stress. However, since each cell contains hundreds not quite identical copies of IGS, the structure and functions of this locus remain poorly understood, and the dynamics of its products has not been specially studied. In this work, we used quantitative PCR to measure the expression levels of various rDNA regions at different times after inhibition of the transcription by Actinomycin D applied in high doses. This approach allowed us to measure real or extrapolated half-life times of some IGS loci. Our study reveals characteristic dynamic patterns suggestive of various pathways of RNA utilization and decay.


Assuntos
DNA Espaçador Ribossômico/metabolismo , RNA/biossíntese , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , Células HeLa , Humanos , RNA/análise , RNA/genética , RNA/isolamento & purificação
20.
Mycologia ; 111(5): 719-729, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31348716

RESUMO

Corn bins in the midwestern United States can reach temperatures up to 52 C. High temperatures combined with sufficient moisture and humidity in bins provide the perfect environment to promote the growth of thermophilic and thermotolerant fungi. In this article, we characterize for the first time thermophilic and thermotolerant fungi in corn grain bins using culture-based methods and pyrosequencing techniques. Corn samples were collected from local farms in western Illinois. Samples were plated and incubated at 50 C using a variety of approaches. Of several hundred kernels examined, more than 90% showed colonization. Species identified using culture methods included Thermomyces lanuginosus, Thermomyces dupontii, Aspergillus fumigatus, Thermoascus crustaceus, and Rhizomucor pusillus. Pyrosequencing was also performed directly on corn grain using fungal-specific primers to determine whether thermophilic fungi could be detected using this technique. Sequences were dominated by pathogenic fungi, and thermophiles were represented by less than 2% of the sequences despite being isolated from 90% of the grain samples using culturing techniques. The high abundance of previously undocumented viable fungi in corn could have negative implications for grain quality and pose a potential risk for workers and consumers of corn-derived products in the food industry. Members of the Sordariales were absent among thermophile isolates and were not represented in nuc rDNA internal transcribed spacer (ITS) sequences. This is in striking contrast with results obtained with other substrates such as litter, dung, and soils, where mesophilic and thermophilic members of the Sordariaceae and Chaetomiaceae are common. This absence appears to reflect an important difference between the ecology of Sordariales and other orders within the Ascomycota in terms of their ability to compete in microhabitats rich in sugars and living tissues.


Assuntos
Fungos/classificação , Fungos/isolamento & purificação , Temperatura Alta , Micobioma , Grãos Integrais/microbiologia , Zea mays/microbiologia , Contagem de Colônia Microbiana , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , Fungos/crescimento & desenvolvimento , Fungos/efeitos da radiação , Illinois , Técnicas Microbiológicas , Filogenia , Análise de Sequência de DNA
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA