Your browser doesn't support javascript.
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 10.024
Filtrar
1.
J Parasitol ; 106(1): 71-81, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31995717

RESUMO

An unusual coccidian parasite was described previously from the prostate of a male Antechinus flavipes (family: Dasyuridae; common name: yellow-footed antechinus). Morphometrics and a partial nuclear 18S small subunit rDNA (18S rDNA) sequence were used to assign this parasite to the genus Eimeria; it was named Eimeria taggarti. We generated full nuclear 18S rDNA and mitochondrial genome sequences from this parasite and used the newly completed 18S rDNA and mitochondrial cytochrome c oxidase subunit I (COI) sequences to perform a more in-depth phylogenetic analysis. The parasite clustered closely with Choleoeimeria spp. and Acroeimeria spp. infecting herptiles in a well-supported clade that was the sister lineage to the Eimeriidae sensu stricto. The mitochondrial genome of this parasite contained 2 inverted segments compared to mitochondrial genomes from parasites in the Eimeriidae sensu stricto (i.e., Stieda body-possessing coccidia with 4 dizoic sporocysts); this mitochondrial genome arrangement was shared with the only Choleoeimeria species for which sequence data were available publicly. Examination of histological preparations and TEM images uncovered bivalvate sporocysts and otherwise confirmed previously described morphological features of the parasite. Based on our phylogenetic analyses and histological observations, we propose the generic reclassification of E. taggarti to Choleoeimeria taggarti n. comb.


Assuntos
Coccidiose/veterinária , Eimeriidae/genética , Genoma Mitocondrial/genética , Marsupiais/parasitologia , Próstata/parasitologia , Animais , Coccidiose/parasitologia , DNA de Protozoário/química , DNA Ribossômico/química , Eimeriidae/classificação , Eimeriidae/isolamento & purificação , Eimeriidae/ultraestrutura , Complexo IV da Cadeia de Transporte de Elétrons/genética , Masculino , Anotação de Sequência Molecular , Oocistos/ultraestrutura , Filogenia , Reação em Cadeia da Polimerase , RNA de Protozoário/genética , RNA Ribossômico 18S/genética , Alinhamento de Sequência
2.
Parasit Vectors ; 12(1): 515, 2019 Nov 04.
Artigo em Inglês | MEDLINE | ID: mdl-31685003

RESUMO

BACKGROUND: Strongyloidiasis is a health problem in Vietnam, but appropriate information is still limited. The aim of this study was to determine the prevalence, geographical distribution, epidemiological aspects, symptoms and other health indicators of Strongyloides stercoralis infections in patients from 27 provinces of northern Vietnam attending the Hanoi Medical University Hospital during 2016 and 2017. METHODS: Blood samples of 2000 patients were analyzed for S. stercoralis infection with an IgG ELISA test. Seroprevalence was analyzed by gender, age group, locality of origin (rural or urban areas) and symptoms. Stools from the seropositive patients were examined for the detection of worms which were subsequently used for species identification by morphology and rDNA ITS1 sequencing. RESULTS: A seroprevalence of 20% was detected, showing an increasing prevalence from young to older age groups but without significant gender difference. Seroprevalence was higher in rural areas than in urban areas, both in general and individually in all provinces without exception, and lower in the mountainous areas than in the large valley lowlands. The follow-up of the 400 patients showed eosinophilia in 100% of cases, diarrhoea in 64.5%, digestion difficulties in 58.0%, stomachache in 45.5%, stomach and duodenal ulcers in 44.5%, itching in 28.0% and fever in 9.5%. The prevalence of symptoms and signs were also higher in older age groups than in younger age groups. Worms were detected in stools of 10.5% of the patients. Sequencing of a 501-bp nuclear ribosomal DNA ITS1 fragment allowed for the verification of infection by Strongyloides stercoralis. CONCLUSIONS: To our knowledge, this study is the largest survey of human strongyloidiasis in Vietnam so far and the first molecular identification of this nematode species in this country. Long-term chronicity may probably be usual in infected subjects, mainly in the older age groups.


Assuntos
Strongyloides stercoralis/genética , Estrongiloidíase/epidemiologia , Adolescente , Adulto , Distribuição por Idade , Idoso , Animais , Anticorpos Anti-Helmínticos/sangue , DNA de Helmintos/análise , DNA de Helmintos/química , DNA Ribossômico/análise , DNA Ribossômico/química , Diarreia/epidemiologia , Diarreia/parasitologia , Eosinofilia/epidemiologia , Eosinofilia/parasitologia , Feminino , Geografia , Humanos , Imunoglobulina G/sangue , Enteropatias Parasitárias/epidemiologia , Enteropatias Parasitárias/parasitologia , Masculino , Pessoa de Meia-Idade , Prevalência , População Rural , Estudos Soroepidemiológicos , Strongyloides stercoralis/imunologia , Strongyloides stercoralis/isolamento & purificação , Estrongiloidíase/parasitologia , Vietnã/epidemiologia , Adulto Jovem
3.
Vet Parasitol ; 276: 108965, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31726324

RESUMO

Tritrichomonas foetus isolates from feline and bovine origin has been previously shown to carry a certain degree of genetic heterogeneity. Here, novel candidate molecular markers were developed by means of multilocus sequence typing of the gap2 gene (encoding for T. foetus glyceraldehyde-3-phosphate dehydrogenase), ITS region, the TR7/TR8 variable-length repeat and microsatellite genotyping. These markers were used to characterize T. foetus field isolates from bulls and domestic cats and to compare phylogenetically with the following ATCC isolates: T. foetus isolated from cattle and pig (syn. Tritrichomonas suis), Tritrichomonas mobilensis, Tetratrichomonas gallinarum and Pentatrichomonas hominis. Among them, TFMS10 and TFMS7 were found to be the most polymorphic markers. Moreover, an 809 bp fragment of the gap2 gene was successfully amplified from all the trichomonads included in this study and the sequence analysis revealed differences between T. foetus porcine and feline genotypes and T. mobilensis in comparison to the bovine T. foetus ATCC isolate. The TR7/TR8 repeat pattern was not reproducible, being only consistent the fragments of approximately 110 and 217 bp. Sequence analysis of the latter revealed the existence of 3 SNPs resulting in 98.6 % homology between bovine and feline isolates. A search for similar sequences was carried out to develop a Restriction Length Fragment Polymorphism analysis. A 503 bp region, named TF1, revealed the existence of two BbvI restriction enzyme sites that were able to generate different length fragments for T. foetus feline and bovine isolates. Finally, the neighbour-joining analyses showed that T. foetus porcine genotype clusters together with bovine genotype, whereas T. mobilensis and the feline genotype form a separate cluster.


Assuntos
Doenças do Gato/parasitologia , Doenças dos Bovinos/parasitologia , Marcadores Genéticos , Infecções Protozoárias em Animais/parasitologia , Tritrichomonas foetus/genética , Animais , Sequência de Bases , Gatos , Bovinos , Sequência Consenso , DNA Ribossômico/química , Gliceraldeído-3-Fosfato Desidrogenase (Fosforiladora)/genética , Masculino , Repetições de Microssatélites/genética , Repetições Minissatélites , Dados de Sequência Molecular , Tipagem de Sequências Multilocus/veterinária , Filogenia , Polimorfismo de Fragmento de Restrição , Polimorfismo de Nucleotídeo Único , Sequências Repetitivas de Ácido Nucleico/genética , Alinhamento de Sequência/veterinária , Tritrichomonas foetus/classificação
4.
Genes Dev ; 33(23-24): 1688-1701, 2019 12 01.
Artigo em Inglês | MEDLINE | ID: mdl-31727772

RESUMO

Human nucleolar organizer regions (NORs), containing ribosomal gene (rDNA) arrays, are located on the p-arms of acrocentric chromosomes (HSA13-15, 21, and 22). Absence of these p-arms from genome references has hampered research on nucleolar formation. Previously, we assembled a distal junction (DJ) DNA sequence contig that abuts rDNA arrays on their telomeric side, revealing that it is shared among the acrocentrics and impacts nucleolar organization. To facilitate inclusion into genome references, we describe sequencing the DJ from all acrocentrics, including three versions of HSA21, ∼3 Mb of novel sequence. This was achieved by exploiting monochromosomal somatic cell hybrids containing single human acrocentric chromosomes with NORs that retain functional potential. Analyses revealed remarkable DJ sequence and functional conservation among human acrocentrics. Exploring chimpanzee acrocentrics, we show that "DJ-like" sequences and abutting rDNA arrays are inverted as a unit in comparison to humans. Thus, rDNA arrays and linked DJs represent a conserved functional locus. We provide direct evidence for exchanges between heterologous human acrocentric p-arms, and uncover extensive structural variation between chromosomes and among individuals. These findings lead us to revaluate the molecular definition of NORs, identify novel genomic structural variation, and provide a rationale for the distinctive chromosomal organization of NORs.


Assuntos
Cromossomos/química , Cromossomos/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Região Organizadora do Nucléolo/química , Região Organizadora do Nucléolo/genética , Animais , Sequência de Bases , Linhagem Celular , Sequência Conservada/genética , Estruturas Genéticas/genética , Variação Genética , Humanos , Células Híbridas , Camundongos , Pan troglodytes/genética
5.
Korean J Parasitol ; 57(4): 439-444, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31533413

RESUMO

Since Kudoa septempuntata was identified as a causative agent of food poisoning associated with raw olive flounder Paralichthys olivaceus, interest and concern regarding the parasite have increased. However, there have been no investigations or reports of other Kudoa species infecting the fish (except for K. paralichthys, which infects the brain) in Korea. We found cysts filled with myxospores of Kudoa species in muscles of cultured olive flounder specimens and identified these to the species level. Mature spores were quadrate, measuring 8.7±0.5 µm in length, 9.2±0.4 µm in thickness, and 12.9±0.6 µm in width. The spores containing 4 polar capsules had a length of 2.1±0.2 µm and a width of 1.8±0.3 µm. The partial 18S and 28S rDNA of isolates showed 99-100% similarities with K. ogawai. Using these morphological and molecular analyses, the species was identified as K. ogawai. This study is the first report of K. ogawai infection in cultured olive flounder in Korea.


Assuntos
Doenças dos Peixes/parasitologia , Linguado/parasitologia , Myxozoa/isolamento & purificação , Doenças Parasitárias em Animais/parasitologia , Animais , Sequência de Bases , DNA Ribossômico/química , Pesqueiros , Músculos/parasitologia , Myxozoa/genética , RNA Ribossômico 18S/genética , RNA Ribossômico 28S/genética , República da Coreia
6.
Biocontrol Sci ; 24(3): 129-136, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31527343

RESUMO

Previously, we isolated and examined a bacterial strain designated as TM-I-3, belonging to the genus Bacillus, from soil in Nagasaki, Japan. This bacterium was able to inhibit the growth of molds, without coming into direct contact with them. Non-contact antifungals are capable of providing multidirectional inhibition and may contribute to disease prevention. In this study, we revealed the bacteriological properties of TM-I-3 and evaluated the antifungal activity of the compounds emitted from this bacterium. In addition, we analyzed the antimicrobial substances released from TM-I-3 using GC/MS to elucidate the mechanism of its action. Antimicrobial compounds from strain TM-I-3 were identified as acetic acid, propanoic acid, isovaleric acid, 2-methylbutanoic acid, and benzaldehyde, which are all reported to have antimicrobial activity. TM-I-3 demonstrated possible efficacy in inhibiting the growth of Aspergillus fumigatus, Cladosporium cladosporioides and Penicillium expansum, which may lead to inhibition of common fungal contaminants of household products and prevention of some pulmonary diseases.


Assuntos
Antifúngicos/farmacologia , Aspergillus fumigatus/efeitos dos fármacos , Bacillus/metabolismo , Cladosporium/efeitos dos fármacos , Penicillium/efeitos dos fármacos , Compostos Orgânicos Voláteis/farmacologia , Antifúngicos/química , Antifúngicos/metabolismo , Aspergillus fumigatus/crescimento & desenvolvimento , Bacillus/classificação , Bacillus/genética , Cladosporium/crescimento & desenvolvimento , Análise por Conglomerados , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Cromatografia Gasosa-Espectrometria de Massas , Japão , Penicillium/crescimento & desenvolvimento , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Compostos Orgânicos Voláteis/química , Compostos Orgânicos Voláteis/metabolismo
7.
Artigo em Inglês | MEDLINE | ID: mdl-31510040

RESUMO

Extracellular DNA (exDNA) pool in aquatic environments is a valuable source for biomonitoring and bioassessment. However, degradation under particular environmental conditions can hamper exDNA detectability over time. In this study, we analyzed how different biotic and abiotic factors affect the degradation rate of extracellular environmental DNA using 16S rDNA sequences extracted from the sediment of a eutrophic lake and Anabaena variabilis cultured in the laboratory. We exposed the extracted exDNA to different levels of temperature, light, pH, and bacterial activity, and quantitatively analyzed the concentration of exDNA during 4 days. The solution containing bacteria for microbial activity treatment was obtained from the lake sediment using four consecutive steps of filtration; two mesh filters (100 µm and 60 µm mesh) and two glass fiber filters (2.7 µm and 1.2 µm pore-sized). We found that temperature individually and in combination with bacterial abundance had significant positive effects on the degradation of exDNA. The highest degradation rate was observed in samples exposed to high microbial activity, where exDNA was completely degraded within 1 day at a rate of 3.27 day-1. Light intensity and pH had no significant effects on degradation rate of exDNA. Our results indicate that degradation of exDNA in freshwater ecosystems is driven by the combination of both biotic and abiotic factors and it may occur very fast under particular conditions.


Assuntos
DNA Ribossômico/análise , Lagos/microbiologia , Anabaena variabilis/metabolismo , Biodegradação Ambiental , DNA Ribossômico/química , DNA Ribossômico/efeitos da radiação , Eutrofização , Sedimentos Geológicos/microbiologia , Concentração de Íons de Hidrogênio , Luz , Temperatura Ambiente , Microbiologia da Água
8.
Appl Microbiol Biotechnol ; 103(19): 8241-8253, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31482282

RESUMO

While numerous reports exist on the axenic culturing of different hydrogen-oxidizing bacteria (HOB), knowledge about the enrichment of microbial communities growing on hydrogen, oxygen, and carbon dioxide as sole carbon and energy sources remains negligible. We want to elucidate if in such enrichments, most enriched populations are HOBs or heterotrophic organisms. In the present study, bacteria enriched from a soil sample and grown over 5 transfers using a continuous supply of hydrogen, oxygen, and carbon dioxide to obtain an enriched autotrophic hydrogen-oxidizing microbiome. The success of the enrichment was evaluated by monitoring ammonium consumption and biomass concentration for 120 days. The shift in the microbial composition of the original soil inoculum and all transfers was observed based on 16S rRNA amplicon sequencing. The hydrogen-oxidizing facultative chemolithoautotroph Hydrogenophaga electricum was isolated and found to be one of the abundant species in most transfers. Moreover, Achromobacter was isolated both under heterotrophic and autotrophic conditions, which was characterized as a hydrogen-oxidizing bacterium. The HOB enrichment condition constructed in this study provided an environment for HOB to develop and conquer in all transfers. In conclusion, we showed that enrichments on hydrogen, oxygen, and carbon dioxide as sole carbon and energy sources contain a diverse mixture of HOB and heterotrophs that resulted in a collection of culturable isolates. These isolates can be useful for further investigation for industrial applications.


Assuntos
Bactérias/classificação , Bactérias/metabolismo , Hidrogênio/metabolismo , Microbiologia do Solo , Compostos de Amônio/metabolismo , Bactérias/genética , Técnicas Bacteriológicas , Dióxido de Carbono/metabolismo , Análise por Conglomerados , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Metagenômica , Oxirredução , Oxigênio/metabolismo , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
9.
Int. microbiol ; 22(3): 325-336, sept. 2019. mapas, graf, tab
Artigo em Inglês | IBECS | ID: ibc-184839

RESUMO

Soil microbial communities are an important component of biological diversity and terrestrial ecosystems which is responsible for processes such as decomposition, mineralization of nutrients, and accumulation of organic matter. One of the factors that provide information on the mechanisms regulating biodiversity is spatial scaling. We characterized the microbial communities using 16S rRNA gene sequences from DNA isolated from halite at various locations and correlated these to geographic distance in the Uyuni salt flat (Bolivia). Sequences from each site were analyzed to determine any spatial patterns of diversity, as well as to describe the microbial communities. Results suggest that different taxa are able to disperse over Uyuni's surface crust regardless of distance. As expected, ubiquitous taxa included members of Halobacteriaceae such as Haloarcula, Halorubrum, Halorhabdus, Halolamina, and halophilic bacteria Salinibacter, Halorhodospira, and unclassified members of the Gammaproteobacteria. Archaeal communities were homogeneous across the salt flat. In contrast, bacterial communities present strong local variations which could be attributed to external factors. Likely sources for these variations are the Rio Grande river influent in the south shore and the Tunupa volcano influencing the northern area


No disponible


Assuntos
Microbiologia do Solo , Bactérias/classificação , Biota , Archaea/classificação , RNA Ribossômico 16S/genética , Bolívia , Bactérias/genética , Archaea/genética , Análise por Conglomerados , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , DNA Arqueal/química , DNA Arqueal/genética , Metagenômica , Filogenia
10.
Int. microbiol ; 22(3): 343-353, sept. 2019. ilus, graf, tab
Artigo em Inglês | IBECS | ID: ibc-184841

RESUMO

The objective of the present study was to isolate Actinobacteria, preferably Streptomyces spp. from the rhizosphere soils of three ethno-medicinal plants collected in Serbia (Papaver rhoeas, Matricaria chamomilla, and Urtica dioica) and to screen their antifungal activity against Candida spp. Overall, 103 sporulating isolates were collected from rhizosphere soil samples and determined as Streptomyces spp. Two different media and two extraction procedures were used to facilitate identification of antifungals. Overall, 412 crude cell extracts were tested against Candida albicans using disk diffusion assays, with 42% (43/103) of the strains showing the ability to produce antifungal agents. Also, extracts inhibited growth of important human pathogens: Candida krusei, Candida parapsilosis, and Candida glabrata. Based on the established degree and range of antifungal activity, nine isolates, confirmed as streptomycetes by 16S rRNA sequencing, were selected for further testing. Their ability to inhibit Candida growth in liquid culture, to inhibit biofilm formation, and to disperse pre-formed biofilms was assessed with active concentrations from 8 to 250 μg/mL. High-performance liquid chromatographic profiles of extracts derived from selected strains were recorded, revealing moderate metabolic diversity. Our results proved that rhizosphere soil of ethno-medicinal plants is a prolific source of streptomycetes, producers of potentially new antifungal compounds


No disponible


Assuntos
Antifúngicos/metabolismo , Candida/efeitos dos fármacos , Rizosfera , Microbiologia do Solo , Streptomyces/isolamento & purificação , Candida/crescimento & desenvolvimento , RNA Ribossômico 16S/genética , Sérvia , Streptomyces/classificação , Streptomyces/genética , Análise por Conglomerados , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética
11.
Int. microbiol ; 22(3): 377-390, sept. 2019. mapas, graf, tab
Artigo em Inglês | IBECS | ID: ibc-184845

RESUMO

Easter Island is an isolated volcanic island in the Pacific Ocean. Despite the extended knowledge about its origin, flora, and fauna, little is known about the bacterial diversity inhabiting this territory. Due to its isolation, Easter Island can be considered as a suitable place to evaluate microbial diversity in a geographically isolated context, what could shed light on actinobacterial occurrence, distribution, and potential novelty. In the present study, we performed a comprehensive analysis of marine Actinobacteria diversity of Easter Island by studying a large number of coastal sampling sites, which were inoculated into a broad spectrum of different culture media, where most important variations in composition included carbon and nitrogen substrates, in addition to salinity. The isolates were characterized on the basis of 16S ribosomal RNA gene sequencing and phylogenetic analysis. High actinobacterial diversity was recovered with a total of 163 pure cultures of Actinobacteria representing 72 phylotypes and 20 genera, which were unevenly distributed in different locations of the island and sample sources. The phylogenetic evaluation indicated a high degree of novelty showing that 45% of the isolates might represent new taxa. The most abundant genera in the different samples were Micromonospora, Streptomyces, Salinispora, and Dietzia. Two aspects appear of primary importance in regard to the high degree of novelty and diversity of Actinobacteria found. First, the application of various culture media significantly increased the number of species and genera obtained. Second, the geographical isolation is considered to be of importance regarding the actinobacterial novelty found


No disponible


Assuntos
Actinobacteria/classificação , Actinobacteria/isolamento & purificação , Biodiversidade , Microbiologia Ambiental , Actinobacteria/genética , Técnicas Bacteriológicas , Chile , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Filogenia , Polinésia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
12.
Appl Microbiol Biotechnol ; 103(19): 8157-8168, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31401751

RESUMO

Chlorogenic acids (CGA), the most abundant natural polyphenol present in human diet and plants, have attracted considerable research interest because of their broad bioactivities including the antimicrobial activity. However, little is known about their influences on intestinal bacterial communities. Here, we described a response in intestinal microbiome to CGA using a porcine model. Twenty-four weaned pigs were allotted to two groups and fed with a basal diet or a basal diet containing 1000 mg/kg CGA. Results showed that CGA significantly increased the length of the small intestine (P < 0.05) and enhanced the activity of diamine oxidase (DAO) and the concentration of MHC-II in the jejunal and ileal mucosa (P < 0.05). Moreover, the acetate concentration in ileum and cecum digesta, and the propionate and butyrate concentrations in the cecum digesta, were significantly elevated by CGA (P < 0.05). Interestingly, CGA significantly increased the total 16S rRNA gene copies and bacterial alpha diversity in the cecum (P < 0.05). The relative abundance of bacteria from phyla Firmicutes and Bacteroidetes was increased in the cecum digesta (P < 0.05), whereas the abundance of bacteria from phylum Protebacteria was decreased by CGA (P < 0.05). Importantly, pigs on CGA-containing diet had higher abundance of Lactobacillus spp., Prevotella spp., Anaerovibrio spp., and Alloprevotella spp. in the cecum (P < 0.05). Not only did our study suggest a synergic response of intestinal barrier function and microbiota to the CGA, but the result will also contribute to understanding of the mechanisms behind the CGA-modulated gut health.


Assuntos
Anti-Infecciosos/administração & dosagem , Bactérias/efeitos dos fármacos , Ácido Clorogênico/administração & dosagem , Microbioma Gastrointestinal/efeitos dos fármacos , Animais , Bactérias/classificação , Bactérias/genética , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Conteúdo Gastrointestinal/química , Metagenômica , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Suínos
13.
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
14.
World J Microbiol Biotechnol ; 35(9): 132, 2019 Aug 20.
Artigo em Inglês | MEDLINE | ID: mdl-31432260

RESUMO

This paper aims to characterize halophilic bacteria inhabiting Algerian Saline Ecosystems (Sebkha and Chott) located in arid and semi-arid ecoclimate zones (Northeastern Algeria). In addition, screening of enzymatic activities, heavy metal tolerance and antagonistic potential against phytopathogenic fungi were tested. A total of 74 bacterial isolates were screened and phylogenetically characterized using 16S rRNA gene sequencing. The results showed a heterogeneous group of microorganisms falling within two major phyla, 52 strains belonging to Firmicutes (70.2%) and 22 strains (30.8%) of γ-Proteobacteria. In terms of main genera present, the isolates were belonging to Bacillus, Halobacillus, Lentibacillus, Oceanobacillus, Paraliobacillus, Planomicrobium, Salicola, Terribacillus, Thalassobacillus, Salibacterium, Salinicoccus, Virgibacillus, Halomonas, Halovibrio, and Idiomarina. Most of the enzymes producers were related to Bacillus, Halobacillus, and Virgibacillus genera and mainly active at 10% of growing salt concentrations. Furthermore, amylase, esterase, gelatinase, and nuclease activities ranked in the first place within the common hydrolytic enzymes. Overall, the isolates showed high minimal inhibitory concentration values (MIC) for Ni2+ and Cu2+ (0.625 to 5 mM) compared to Cd2+ (0.1 to 2 mM) and Zn2+ (0.156 to 2 mM). Moreover, ten isolated strains belonging to Bacillus, Virgibacillus and Halomonas genera, displayed high activity against the pathogenic fungi (Botrytis cinerea, Fusarium oxyporum, F. verticillioides and Phytophthora capsici). This study on halophilic bacteria of unexplored saline niches provides potential sources of biocatalysts and novel bioactive metabolites as well as promising candidates of biocontrol agents and eco-friendly tools for heavy metal bioremediation.


Assuntos
Antibiose , Bactérias/isolamento & purificação , Bactérias/metabolismo , Biota , Microbiologia Ambiental , Salinidade , Argélia , Bactérias/classificação , Bactérias/genética , Análise por Conglomerados , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Fungos/crescimento & desenvolvimento , Hidrolases/análise , Metais Pesados/metabolismo , Metais Pesados/toxicidade , Testes de Sensibilidade Microbiana , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
15.
Antonie Van Leeuwenhoek ; 112(10): 1533-1544, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31165292

RESUMO

A taxonomic study of a Gram-stain negative, rod-shaped, motile, asporogenous, catalase- and oxidase-positive bacterium, sh-6T, forming pink-red colonies, isolated from a contaminated R2A plate in the laboratory was performed. Its optimum growth temperature was determined to be 28 °C in the absence of NaCl on R2A plates. On the basis of 16S rRNA gene sequence analysis, strain sh-6T belongs to the genus Hymenobacter and is closely related to Hymenobacter deserti ZLB-3T (95.05%), Hymenobacter paludis KBP-30T (94.96%), Hymenobacter coalescens WW84T (94.04%), Hymenobacter gummosus ANT-18T (93.38%), Hymenobacter ocellatus Myx2105T (93.70%), Hymenobacter jeollabukensis 1-3-3-8T (93.48%) and Hymenobacter koreensis GYR3077T (93.21%). Comparison of the genome of strain sh-6T and that of H. gummosus ANT-18T gave digital DNA-DNA hybridization and Average Nucleotide Identity values of 20.6% and 78.4%, respectively. The respiratory isoprenoid quinone and polyamine component were identified as MK-7 and sym-homospermidine, respectively. The major cellular fatty acids identified as iso-C15:0, summed feature 4 (iso-C17:1 I/anteiso B), iso-C16:0, iso-C17:0 3-OH and iso-C17:0. The major polar lipid of strain sh-6T determined to be phosphatidylethanolamine. The DNA G+C content was determined to be 60.5 mol%. On the basis of the evidence presented in this study, a novel species of the genus Hymenobacter, Hymenobacter oligotrophus sp. nov., is proposed, with the type strain sh-6T (= CCTCC AB 2016064T = KCTC 62345T).


Assuntos
Bacteroidetes/classificação , Bacteroidetes/isolamento & purificação , Microbiologia Ambiental , Ágar , Técnicas de Tipagem Bacteriana , Bacteroidetes/genética , Bacteroidetes/fisiologia , Composição de Bases , Análise por Conglomerados , Meios de Cultura , Citosol/química , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Ácidos Graxos/análise , Locomoção , Hibridização de Ácido Nucleico , Fosfolipídeos/análise , Filogenia , Quinonas/análise , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Cloreto de Sódio/metabolismo , Temperatura Ambiente
16.
Appl Microbiol Biotechnol ; 103(15): 6353-6367, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31161391

RESUMO

Anaerobic digestion is an important biotechnology treatment process for conversion of waste to energy. In this study, a comparative core microbiome approach, i.e., determining taxa that are shared in functioning digesters but not shared in non-functioning digesters, was used to determine microbial taxa that could play key roles for effective anaerobic digestion. Anaerobic digester functions were impaired by adding the broad-spectrum antimicrobial triclosan (TCS) or triclocarban (TCC) at different concentrations, and the core microbiomes in both functioning and non-functioning anaerobic digesters were compared. Digesters treated with high (2500 mg/kg) or medium (450 mg/kg) TCS and high (850 mg/kg) TCC concentrations lost their function, i.e., methane production decreased, effluent volatile fatty acid concentrations increased, and pH decreased. Changes in microbial community diversity and compositions were assessed using 16S rRNA gene amplicon sequencing. Microbial richness decreased significantly in non-functioning digesters (p < 0.001). Microbial community compositions in non-functioning digesters significantly differed from those in functioning digesters (p = 0.001, ANOSIM). Microbes identified as potentially key taxa included previously known fatty acid-degrading syntrophs and amino acid-degrading syntrophs. A diverse group of syntrophs detected in this study had low relative abundance in functioning digesters, suggesting the importance of rare microbes in anaerobic digester operation. The comparative microbiome approach used in this study can be applied to other microbial systems where a community-driven biological phenomena can be observed directly.


Assuntos
Biota , Metano/metabolismo , Esgotos/microbiologia , Purificação da Água , Anaerobiose , Análise por Conglomerados , DNA Ribossômico/química , DNA Ribossômico/genética , Ácidos Graxos Voláteis/metabolismo , Concentração de Íons de Hidrogênio , Metagenômica , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
17.
Jpn J Infect Dis ; 72(5): 343-346, 2019 Sep 19.
Artigo em Inglês | MEDLINE | ID: mdl-31155602

RESUMO

We performed Leptospira culture analysis of 76 clinical samples collected from animals and of six soil samples for the investigation of a leptospirosis outbreak in southern Thailand in 2017. Leptospires were recovered from a kidney sample (a fatal canine leptospirosis case) and from all the soil samples. Next, 16S rRNA sequence analysis demonstrated that the clinical isolate was closely related to the pathogenic L. interrogans, whereas the soil isolates represented different species, including pathogenic L. ellisii, intermediate L. wolffii, and nonpathogenic L. yanagawae. Multilocus sequence typing identified an isolate of L. interrogans as a novel sequence type (ST263), suggesting that the causative agent of the canine leptospirosis in the southern Thailand outbreak has a unique genetic profile.


Assuntos
Doenças dos Animais/epidemiologia , Doenças dos Animais/microbiologia , Surtos de Doenças , Genótipo , Leptospira interrogans/classificação , Leptospira interrogans/isolamento & purificação , Leptospirose/veterinária , Animais , Animais Domésticos , Análise por Conglomerados , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Leptospira interrogans/genética , Leptospirose/epidemiologia , Leptospirose/microbiologia , Tipagem de Sequências Multilocus , Filogenia , RNA Ribossômico 16S/genética , Ratos , Microbiologia do Solo , Tailândia/epidemiologia
18.
Antonie Van Leeuwenhoek ; 112(10): 1559-1565, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31172329

RESUMO

A Gram-stain negative, non-motile, and yellow-coloured bacterium, designated 17J28-26T, was isolated from soil in Jeju Island, Korea. Phylogenetic analyses based on 16S rRNA gene sequences revealed that strain 17J28-26T formed a distinct lineage within the family Chitinophagaceae (order Chitinophagales, class Chitinophagia), and is closely related to Flavisolibacter ginsenosidimutans (96.8% 16S rRNA gene sequence similarity), and Flavisolibacter ginsengisoli (96.6%). Growth was observed at 18-37 °C (optimum 30 °C) in R2A medium at pH 7.0. The major cellular fatty acids of strain 17J28-26T were summed feature 3 (C16:1 ω6c and/or C16:1 ω7c), and iso-C15:0. The predominant respiratory quinone was MK-7. The major polar lipid was identified as phosphatidylethanolamine. Based on biochemical, chemotaxonomic and phylogenetic characteristics, strain 17J28-26T represents a novel bacterial species within the family Chitinophagaceae, for which the name Flavisolibacter galbus sp. nov. is proposed. The type strain of Flavisolibacter galbus is 17J28-26T (= KCTC 62222T = JCM 33203T).


Assuntos
Bacteroidetes/classificação , Bacteroidetes/isolamento & purificação , Microbiologia do Solo , Bacteroidetes/genética , Bacteroidetes/fisiologia , Análise por Conglomerados , Citosol/química , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Ácidos Graxos/análise , Concentração de Íons de Hidrogênio , Ilhas , Coreia (Geográfico) , Locomoção , Fosfolipídeos/análise , Filogenia , Pigmentos Biológicos/metabolismo , Quinonas/análise , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Temperatura Ambiente
19.
PLoS Negl Trop Dis ; 13(6): e0007361, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-31173595

RESUMO

The composition and structure of microbial communities associated with mosquitoes remain poorly understood despite their important role in host biology and potential to be harnessed as novel strategies for mosquito-borne disease control. We employed MiSeq sequencing of the 16S rRNA gene amplicons to characterize the bacterial flora of field-collected populations of Aedes mcintoshi and Aedes ochraceus, the primary vectors of Rift Valley fever (RVF) virus in Kenya. Proteobacteria (53.5%), Firmicutes (22.0%) and Actinobacteria (10.0%) were the most abundant bacterial phyla accounting for 85.5% of the total sequences. Non-metric multi-dimensional scaling plots based on Bray-Curtis dissimilarities revealed a clear grouping of the samples by mosquito species, indicating that the two mosquito species harbored distinct microbial communities. Microbial diversity, richness and composition was strongly influenced by the site of mosquito collection and overall, Ae. ochraceus had significantly higher microbial diversity and richness than Ae. mcintoshi. Our findings suggest that host species and site of collection are important determinants of bacterial community composition and diversity in RVF virus vectors and these differences likely contribute to the spatio-temporal transmission dynamics of RVF virus.


Assuntos
Aedes/microbiologia , Bactérias/classificação , Bactérias/genética , Microbioma Gastrointestinal , Mosquitos Vetores/microbiologia , Animais , Análise por Conglomerados , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Feminino , Quênia , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
20.
Antonie Van Leeuwenhoek ; 112(10): 1567-1575, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31147966

RESUMO

A novel Gram-negative bacterium, non-motile and short rod-shaped, designated strain GY511T, was isolated from the intestines of fish collected from Maowei Sea, China. Growth occurred at pH 6.0-9.0 (optimum 7.0), 4-37 °C (optimum 28 °C) and at 0-2.5% (w/v) NaCl (optimum 1.0%). The result of 16S rRNA gene sequence analysis showed that strain GY511T is closely related to O. oryzae NBRC 113109T (97.6%), O. konkukae DSM 105395T (97.4%), Ottowia beijingensis CGMCC 1.12324T (95.9%), Ottowia pentelensis DSM 21699T (95.2%) and Ottowia thiooxydans DSM 14619T (95.0%). The DNA-DNA hybridization values of strain GY511T with O. oryzae NBRC 113109T and O. konkukae DSM 105395T were 35.4 ± 3.1% and 26.3 ± 1.8%, respectively. The major fatty acids (> 10%) were identified as summed feature 3 (C16:1ω7c and/or C16:1ω6c), C16:0 and summed feature 8 (C18:1ω7c and/or C18:1ω6c) and the major respiratory quinone was ubiquinone-8 (Q-8). The polar lipids comprised diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylmethylethanolamine, two unidentified aminolipids and an unidentified phospholipid. The G+C content of the genomic DNA was 62.9 mol%. Thiosulfate could be utilized as co-substrate for aerobic growth and was oxidised to sulfate. On the basis of phenotypic, chemotaxonomic and molecular data, strain GY511T is considered to represent a novel species of the genus Ottowia, for which the name Ottowia flava sp. nov. is proposed. The type strain is GY511T (= NBRC 113500T = DSM 107425T = CGMCC 1.13650T).


Assuntos
Comamonadaceae/classificação , Comamonadaceae/isolamento & purificação , Peixes/microbiologia , Intestinos/microbiologia , Aerobiose , Animais , Organismos Aquáticos/microbiologia , Técnicas de Tipagem Bacteriana , Composição de Bases , China , Análise por Conglomerados , Comamonadaceae/genética , Comamonadaceae/fisiologia , Citosol/química , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Ácidos Graxos/análise , Concentração de Íons de Hidrogênio , Hibridização de Ácido Nucleico , Fosfolipídeos/análise , Filogenia , Quinonas/análise , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Cloreto de Sódio/metabolismo , Temperatura Ambiente
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA