RESUMO
The industry of fine wines and also locally consumed table wines is emerging in Brazil with an increasing volume and economic impact. Enologists in this region currently lack information about the prevalence and characteristics of spoilage yeasts, which may contaminate and potentially undervalue Brazilian wines. Herein, we analyzed 50 local red wines including 27 fine wines (V. vinifera) and 23 table wines (V. labrusca). Presumptive spoilage yeasts were isolated on differential medium, and classified by RFLP-PCR and sequencing of the ITS1-5.8S-ITS2 and D1/D2 26S rDNA loci. The prevalence of spoilage yeasts in fine wines (11 %) was comparable to that reported in European and US wines, and significantly lower than that observed for local table wines (70 %). The majority of isolates belonged to Brettanomyces bruxelliensis, followed by Pichia guillermondii, and more rarely Candida wickerhamii and Trigonopsis cantarelli. The Brettanomyces isolates varied greatly in off-flavor production, displayed ethanol tolerance (>10 % by volume), tolerated sulfite (≥0.68 mg/l mSO2), and 39 % of them grew on ethanol as sole carbon source. We discuss the causes and consequences of spoilage yeasts in relation to the Brazilian wine industry.
Assuntos
Saccharomyces/isolamento & purificação , Saccharomyces/metabolismo , Vinho/análise , Vinho/microbiologia , Brasil , 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 , Dados de Sequência Molecular , Filogenia , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , RNA Fúngico/genética , RNA Ribossômico/genética , RNA Ribossômico 5,8S/genética , Saccharomyces/classificação , Saccharomyces/genética , Análise de Sequência de DNARESUMO
Based on sequence alignment, oligonucleotide primers targeting the Aeromonas extracellular lipase gene were developed for PCR detection of member of the genus. A pair of primers designed for conserved regions of the gene amplified a 276 bp sequence in all Aeromonas species and tested strains, but did not have a positive result with other Gram-positive and Gram-negative bacteria, showing high specificity and sensitivity. Selective enrichment in alkaline peptone water, followed by centrifugation, and direct usage of cells suspension as template, detected initial populations of 10 c.f.u. ml⻹. Single-strand conformation polymorphism analysis of the PCR products allowed the characterization of Aeromonas strains with a high discriminatory power (Simpson's index = 0.988). The method presented here provides a useful tool for the rapid detection of Aeromonas and the characterization of Aeromonas isolates.
Assuntos
Aeromonas/genética , Aeromonas/isolamento & purificação , Reação em Cadeia da Polimerase/métodos , Polimorfismo Conformacional de Fita Simples , Aeromonas/classificação , Primers do DNA , DNA Bacteriano/genética , Genes Bacterianos , Lipase/genética , RNA Ribossômico 16S/genética , Sensibilidade e Especificidade , Alinhamento de Sequência/métodos , Análise de Sequência de DNARESUMO
Acanthamoeba are abundant in a wide range of environments, and some species are responsible for cutaneous infections, keratitis, and granulomatous amoebic encephalitis (GAE). The conventional detection and isolation of amoeba from clinical and environmental samples involves sampling and culture on non-nutrient Ágar medium. Although efficient, this system requires several transfers in order to eliminate contaminants, and is not appropriate for the isolation of individual amoeba from samples with a biodiverse community. In this study we propose an alternative method for the isolation of monocystic clones of Acanthamoeba. The propose method involves sampling, enrichment, encystment induction, and direct cysts micromanipulation and culture on Ágar plates.
RESUMO
The purpose of this work was to study the biodegradation of citronellol, citronellal and citronellyl acetate by a soil Pseudomonas mendocina strain (IBPse 105) isolated from a Cymbopogon windelandi field. This strain efficiently used citronellol, citronellal, citronellyl acetate and myrcene as sole source of carbon, but was not able to grow on other 15 monoterpenoids evaluated. Gas chromatography-mass spectrometry (GC-MS) analysis of metabolites accumulation during P. medocina IBPse 105 growth on citronellol showed that this strain uses the citronellol catabolic pathway described for other species of the genus. IBPse 105 degradation of citronellyl acetate initiates by its hydrolysis to citronellol. The mini-Tn5 insertion in mutant IBPse 105-303, impaired in citronellol degradation, but able to grow on citronellal, was located in a homologous of the P. aeruginosa atuB gene, that codifies citronellol deshydrogenase.
Assuntos
Monoterpenos/metabolismo , Pseudomonas mendocina/metabolismo , Biotransformação , Cromatografia Gasosa-Espectrometria de Massas , Hidrólise , Mutagênese , Pseudomonas mendocina/genética , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
ABSTRACT Nematicidal activity of 22 monoterpenoids were evaluated in vitro and in pot experiments. Twenty of the twenty-two monoterpenoids significantly reduced hatching, and 11 reduced J2 mobility of the root-knot nematode Meloidogyne incognita at a concentration of 250 mg/liter. In general, compounds with hydroxyl and carbonyl groups exhibited higher nematicidal activity than other terpenoids. Borneol, carveol, citral, geraniol, and alpha-terpineol showed the highest nematicidal activity among the in vitro tested monoterpenoids. These compounds exhibited a dose dependent effect, and drastically reduced eggs hatching and J2 viability at low concentrations. These monoterpenoids, at 100 and 250 mg/kg concentration, diminished root galling of tomato plants in pot experiments. The results suggest that the selected monoterpenoids, and essential oils with high concentration of these compounds, are potential nematicides against Meloidogyne.
Assuntos
Antinematódeos/farmacologia , Monoterpenos/farmacologia , Tylenchoidea/efeitos dos fármacos , AnimaisRESUMO
Proteus mirabilis is one of the most important pathogens associated with complicated urinary tract infections (acute pyelonephritis, bladder infections, kidney stones) and bacteremia, affecting patients with anatomical abnormalities, immunodeficiency, and long-term urinary catheterization. For epidemiological purposes, various molecular typing methods, such as pulse-field gel electrophoresis (PFGE) or ribotyping, have been developed for this pathogen. However, these methods are labor intensive and time-consuming. We evaluated the discriminatory power of several PCR-based fingerprinting methods (RAPD, ISSR, ERIC-PCR, BOX-PCR and rep-PCR) for P. mirabilis clinical isolates. Typing patterns and clustering analysis indicated that RAPD, BOX-PCR and ERIC-PCR differentiated P. mirabilis strains from Escherichia coli, Hafnia alvei, and Morganella morganii. With the exception of rep-PCR, the methods gave medium to high discriminatory efficiency in P. mirabilis. In general, the results obtained with RAPD, BOX-PCR and ERIC-PCR were in good agreement. We concluded that a combination of ERIC-PCR and BOX-PCR results is a rapid and reliable alternative for discrimination among P. mirabilis clinical isolates, contributing to epidemiological studies.
Assuntos
Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Adulto Jovem , Técnicas de Tipagem Bacteriana/métodos , Impressões Digitais de DNA/métodos , Reação em Cadeia da Polimerase/métodos , Proteus mirabilis/genética , DNA Bacteriano/análise , Proteus mirabilis/classificação , Proteus mirabilis/isolamento & purificação , Reprodutibilidade dos Testes , Adulto JovemRESUMO
Proteus mirabilis is one of the most important pathogens associated with complicated urinary tract infections (acute pyelonephritis, bladder infections, kidney stones) and bacteremia, affecting patients with anatomical abnormalities, immunodeficiency, and long-term urinary catheterization. For epidemiological purposes, various molecular typing methods, such as pulse-field gel electrophoresis (PFGE) or ribotyping, have been developed for this pathogen. However, these methods are labor intensive and time-consuming. We evaluated the discriminatory power of several PCR-based fingerprinting methods (RAPD, ISSR, ERIC-PCR, BOX-PCR and rep-PCR) for P. mirabilis clinical isolates. Typing patterns and clustering analysis indicated that RAPD, BOX-PCR and ERIC-PCR differentiated P. mirabilis strains from Escherichia coli, Hafnia alvei, and Morganella morganii. With the exception of rep-PCR, the methods gave medium to high discriminatory efficiency in P. mirabilis. In general, the results obtained with RAPD, BOX-PCR and ERIC-PCR were in good agreement. We concluded that a combination of ERIC-PCR and BOX-PCR results is a rapid and reliable alternative for discrimination among P. mirabilis clinical isolates, contributing to epidemiological studies.