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1.
Microbiologyopen ; 8(1): e00595, 2019 01.
Artigo em Inglês | MEDLINE | ID: mdl-29504263

RESUMO

The fungal community of six sand samples from Saudi Arabia and Jordan deserts was characterized by culture-independent analysis via next generation sequencing of the 18S rRNA genes and by culture-dependent methods followed by sequencing of internal transcribed spacer (ITS) region. By 18S sequencing were identified from 163 to 507 OTUs per sample, with a percentage of fungi ranging from 3.5% to 82.7%. The identified fungal Phyla were Ascomycota, Basal fungi, and Basidiomycota and the most abundant detected classes were Dothideomycetes, Pezizomycetes, and Sordariomycetes. A total of 11 colonies of filamentous fungi were isolated and cultured from six samples, and the ITS sequencing pointed toward five different species of the class Sordariomycetes, belonging to genera Fusarium (F. redolens, F. solani, F. equiseti), Chaetomium (C. madrasense), and Albifimbria (A. terrestris). The results of this study show an unexpectedly large fungal biodiversity in the Middle East desert sand and their possible role and implications on human health.


Assuntos
Biodiversidade , Clima Desértico , Fungos/classificação , Fungos/isolamento & purificação , Temperatura Alta , 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 , Fungos/genética , Fungos/crescimento & desenvolvimento , Jordânia , Técnicas Microbiológicas , Filogenia , RNA Ribossômico 18S/genética , Arábia Saudita , Análise de Sequência de DNA
2.
Int J Antimicrob Agents ; 52(5): 586-592, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30012442

RESUMO

Enterococcus faecium has become a globally disseminated nosocomial pathogen mainly because of acquisition and diffusion of virulence factors and multidrug resistance determinants, including glycopeptides, which are some of the last resort antimicrobials used to treat more serious infections common in high-risk patients. In this study we investigated and characterized hospital-associated (HA) E. faecium isolates collected at Hue Central Hospital, Vietnam. Our results highlighted the spread among hospital wards of a surprisingly heterogeneous multidrug-resistant E. faecium population comprising five different CC17-related sequence types (STs), of which 46% VREf carry the vanB gene. Whole genome sequencing of selected E. faecium isolates showed that VREf from different STs carried the same chromosomal integrated Tn1549-like transposon, with a highly mutated vanB2-operon, showing an increased level of vancomycin resistance (VanB phenotype) and able, in one isolate, to confer resistance to teicoplanin (VanA incongruent phenotype). Two unusual vanA/vanB2-type strains were detected within the vanB2-type ST17 population, harbouring a Tn1546-vanA-like transposon in pJEG40-like plasmids. Wg-SNPs-based analysis showed the genetic relatedness of VSEf/VREf of the same STs and indicated lateral exchange of the Tn1549-like element among isolates followed by clonal expansion. Microevolution among ST17 isolates, including the vanA/vanB2-type strains, and inter-wards VREf transmission, were highlighted. The use of teicoplanin is strongly discouraged in the study hospital because of the spreading of Tn1549-vanB2 associated to teicoplanin resistance. A rational use of glycopeptides and effective surveillance measures are required to reduce nosocomial VSEF/VREf spread and to avoid the rise of unusual and misleading VREf genotypes.


Assuntos
Proteínas de Bactérias/genética , Carbono-Oxigênio Ligases/genética , Enterococcus faecium/isolamento & purificação , Genótipo , Enterococos Resistentes à Vancomicina/isolamento & purificação , Cromossomos Bacterianos , Infecção Hospitalar/epidemiologia , Infecção Hospitalar/microbiologia , Infecção Hospitalar/transmissão , Elementos de DNA Transponíveis , Transmissão de Doença Infecciosa , Farmacorresistência Bacteriana , Enterococcus faecium/genética , Evolução Molecular , Transferência Genética Horizontal , Infecções por Bactérias Gram-Positivas/epidemiologia , Infecções por Bactérias Gram-Positivas/microbiologia , Infecções por Bactérias Gram-Positivas/transmissão , Hospitais , Humanos , Epidemiologia Molecular , Tipagem Molecular , Mutação , Óperon , Plasmídeos , Polimorfismo de Nucleotídeo Único , Teicoplanina/farmacologia , Vancomicina/farmacologia , Enterococos Resistentes à Vancomicina/classificação , Enterococos Resistentes à Vancomicina/genética , Vietnã/epidemiologia , Sequenciamento Completo do Genoma
3.
Infect Genet Evol ; 52: 19-25, 2017 08.
Artigo em Inglês | MEDLINE | ID: mdl-28434988

RESUMO

In Vietnam, the high prevalence of Helicobacter pylori infection represents a serious health problem. Virulence genes of H. pylori have been associated to increased risk of severe gastrointestinal diseases and the genetic background differs in geographical areas. We investigated cagA and vacA genotypes of H. pylori from dyspeptic patients from central Vietnam and the correlation with clinical outcomes; we also performed sequencing analysis of partial cagA gene. Overall, 84% of strains were cagA-positive, 75% were East-Asian type with a prevalence of vacAs1i1m1 and vacAs1i1m2 genotypes (66.7% and 33.3%, respectively) and 9% were Western type vacAs1i1m1 (n=4) and vacAs1i1m2 (n=4); vacAs1i2m2 (n=4) and vacAs2i2m2 (n=2) genotypes were associated to cagA-negative. Strains from gastric ulcer and cancer were of East-Asian type, while cagA-negative or Western strains were from gastritis and duodenal ulcer. H. pylori strains from gastric ulcer patients were predominantly vacAs1i1m1 compared to other vacA genotypes (p<0.05). East-Asian type strains vacAs1i1m1 or vacAs1i1m2 were found in gastric cancer patients and also in less severe disease. Phylogenetic tree analysis of CagA sequences showed the co-circulation of H. pylori of different geographical origins with Western sequences closer related to Cambodia, one of the entry of Western strains in Southeast-Asia through human migrations. Sequence analysis revealed in two Western type strains a chimeric CagA-3' region with identity with East-Asian CagA suggesting recombination event in the process of evolution among East-Asian and Western H. pylori strains. Moreover, polymorphism in CagA multimerization (CM) motif was observed including new East-Asian CM motifs. In conclusion, we have found in central Vietnam a geographically dependent diversity of cagA genotype, with higher rates of cagA-negative and Western-type strains compared with other nation's parts that can partly explain the lower risk of gastric cancer. The polymorphism of CM motifs may explain the variability of disease manifestations of vacAs1i1m1 and s1i1m2 East-Asian isolates.


Assuntos
Antígenos de Bactérias/genética , Proteínas de Bactérias/genética , Dispepsia/microbiologia , Técnicas de Genotipagem/métodos , Helicobacter pylori/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Evolução Molecular , Feminino , Helicobacter pylori/isolamento & purificação , Humanos , Masculino , Pessoa de Meia-Idade , Filogenia , Análise de Sequência de DNA , Vietnã , Adulto Jovem
4.
Sci Rep ; 5: 16306, 2015 Nov 06.
Artigo em Inglês | MEDLINE | ID: mdl-26542754

RESUMO

Dust particles lifting and discharge from Africa to Europe is a recurring phenomenon linked to air circulation conditions. The possibility that microorganisms are conveyed across distances entails important consequences in terms of biosafety and pathogens spread. Using culture independent DNA-based analyses via next generation sequencing of the 16 S genes from the airborne metagenome, the atmospheric microbial community was characterized and the hypothesis was tested that shifts in species diversity could be recorded in relation to dust discharge. As sampling ground the island of Sardinia was chosen, being an ideal cornerstone within the Mediterranean and a crossroad of wind circulation amidst Europe and Africa. Samples were collected in two opposite coastal sites and in two different weather conditions comparing dust-conveying winds from Africa with a control situation with winds from Europe. A major conserved core microbiome was evidenced but increases in species richness and presence of specific taxa were nevertheless observed in relation to each wind regime. Taxa which can feature strains with clinical implications were also detected. The approach is reported as a recommended model monitoring procedure for early warning alerts in frameworks of biosafety against natural spread of clinical microbiota across countries as well as to prevent bacteriological warfare.


Assuntos
Microbiologia do Ar , Poeira , Clima , Análise por Conglomerados , Mar Mediterrâneo , Filogenia , Análise de Componente Principal , RNA Ribossômico 16S/genética
5.
Int J Food Microbiol ; 215: 31-9, 2015 Dec 23.
Artigo em Inglês | MEDLINE | ID: mdl-26325598

RESUMO

Antimicrobial-resistant non-typhoidal Salmonella (NTS) are an important cause of infection in Africa, but there is a lack of information on their molecular mechanisms of resistance and epidemiology. This study contributes to fill this gap through the characterization by pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST), plasmid profiling and analysis of antibiotic-resistance determinants of 94 Salmonella enterica strains isolated from food in Morocco. PFGE revealed considerable heterogeneity among the strains, showing 32 pulsotypes. MLST of strains representative of the different serovars evidenced 13 sequence types (STs), three of which were newly identified (ST1694, ST1768 and ST1818) and nine not previously reported in Morocco. Thirty-four strains harbored from one to four plasmids, of IncI1 group in S. Mbandaka, IncFIIA in S. Typhimurium, IncL/M in S. Hadar and S. Blockley. For the first time in Morocco an intact Salmonella Genomic Island 1 (SGI1) carrying the resistance genes aadA2, floR, tetG, blaPSE-1 and sul1 was detected in S. Typhimurium DT104. In serovar Hadar resistance to ampicillin, tetracycline and streptomycin was associated to blaTEM-1, tetA and strA genes respectively, whereas one mutation in gyrA (Asp87Asn) and one in parC (Thr54Ser) genes conferred resistance to nalidixic acid. These findings improve the information on foodborne Salmonella in Morocco, evidencing the presence of MDR strains potentially dangerous to humans, and provide useful data for future studies.


Assuntos
Antibacterianos/farmacologia , Microbiologia de Alimentos , Salmonella enterica/efeitos dos fármacos , Salmonella enterica/genética , Eletroforese em Gel de Campo Pulsado , Ilhas Genômicas/genética , Humanos , Testes de Sensibilidade Microbiana , Marrocos , Tipagem de Sequências Multilocus , Ácido Nalidíxico/farmacologia , Plasmídeos/genética , Salmonella enterica/isolamento & purificação
6.
Int J Antimicrob Agents ; 45(3): 244-8, 2015 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-25499186

RESUMO

Antimicrobial resistance in Helicobacter pylori has increased worldwide and has become a major cause of treatment failure in many countries, including Vietnam. It is advisable to perform an antibiogram to provide optimal regimens for H. pylori eradication. This study evaluated the rate of antibiotic resistance to the four commonly used antibiotics against H. pylori at a tertiary care hospital in Central Vietnam and analysed point mutations in genes related to clarithromycin (CLA) and levofloxacin (LFX) resistance. A total of 92 H. pylori strains from gastric biopsy specimens were tested; 42.4% were resistant to CLA (primary, 34.2%; secondary, 73.7%), 41.3% to LFX (primary, 35.6%; secondary, 63.2%), 76.1% to metronidazole (MTZ) and 1.1% to amoxicillin. Multidrug resistance was observed in 56.5% (primary, 50.7%; secondary, 78.9%) of isolates (P<0.05). The rate of resistance to LFX was significantly higher in females than males (P<0.05). Most of the CLA- and LFX-resistant strains harboured resistance-associated mutations, with common positions at A2143G and T2182C in the 23S rRNA gene and at Asn-87 or Asp-91 in GyrA. Minimum inhibitory concentrations (MICs) increased in strains carrying quadruple mutations in their 23S rRNA gene and in strains with Asn-87 GyrA mutation (P<0.05). One high-level LFX-resistant strain (MIC=32mg/L) had new mutations with a combination of N87A, A88N and V65I. High resistance rates to CLA, MTZ and LFX discourage standard and LFX-based triple therapies as first-line treatment in Vietnam.


Assuntos
Antibacterianos/farmacologia , Claritromicina/farmacologia , Farmacorresistência Bacteriana , Helicobacter pylori/efeitos dos fármacos , Levofloxacino/farmacologia , Metronidazol/farmacologia , Adolescente , Adulto , Idoso , DNA Girase/genética , Feminino , Infecções por Helicobacter/microbiologia , Helicobacter pylori/isolamento & purificação , Humanos , Masculino , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Mutação Puntual , RNA Ribossômico 23S/genética , Centros de Atenção Terciária , Vietnã , Adulto Jovem
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