Identificação de bactérias contaminantes de fermento de cachaça por seqüenciamento do gene 16s rDNA

Cachaça is a typical Brazilian liquor produced from the distillation of fermented sugarcane juice mainly by Saccharomyces cerevisiae. Most of the domestic production is artisanal, and producers usually are not concerned regarding microbiological control of the fermentation. This study aimed to characterize the contaminant bacterial community of the yeast used in the production of cachaça in an artisanal still. Four samples were collected, of which one (NA) was used for comparison purposes and was collected one year earlier. The remaining samples were collected at three different periods: at the end of the first day of fermentation (NP), after fifteen days (NS), and thirty days after the same yeast was used (NT). Five hundred and eighty-seven sequences were analyzed from the partial sequencing of the 16S rDNA gene. Sequence analyses revealed the presence of 170 operational taxonomic units (OTUs). Of these, only one was shared among three samples and seventeen were shared between two samples. The remaining 152 OTUs were identified only once in distinct samples indicating that the contaminant bacterial population is highly dynamic along the fermentation process. Statistical analyses revealed differences in bacterial composition among samples. Undescribed species in the literature on yeasts of cachaça were found, such as Weissella cibaria, Leuconostoc citreum, and some species of Lactobacillus, in addition to some unknown bacteria. The community of bacteria in the fermentation process is much more complex than it was previously considered. No previous report is known regarding the use of this technique to determine bacterial contaminants in yeast for the production of cachaça.A cachaça é uma bebida típica brasileira produzida a partir da destilação do caldo de cana-de-açúcar fermentado principalmente por Saccharomyces cerevisiae. Grande parte da produção nacional é artesanal, e não há uma preocupação por parte dos produtores quanto ao controle microbiológico da fermentação. Este trabalho objetivou caracterizar a comunidade bacteriana contaminante do fermento utilizado na produção de cachaça em um alambique artesanal. Foram coletadas quatro amostras, sendo uma (NA) utilizada como efeito comparativo e coletada um ano anterior às demais. As restantes foram coletadas em três diferentes períodos: ao final do primeiro dia de fermentação (NP), após quinze dias (NS) e trinta dias após a utilização do mesmo fermento (NT). Foram analisadas, a partir do seqüenciamento parcial do gene 16S rDNA, 587 seqüências. As análises das seqüências revelaram a presença de 170 unidades taxonômicas operacionais. Destas, 152 foram identificadas uma única vez em diferentes amostras, dezessete foram comuns em pelo menos duas amostras e somente uma foi identificada em três amostras, evidenciando uma grande dinâmica populacional bacteriana durante o processo fermentativo. Análises estatísticas revelaram diferenças na composição bacteriana entre as amostras. Foram encontradas espécies ainda não descritas na literatura em fermentos para a produção de cachaça, como Weissella cibaria, Leuconostoc citreum e algumas espécies de Lactobacillus, além de bactérias não conhecidas. Os resultados revelaram que a comunidade de bactérias contaminantes do processo fermentativo é muito mais complexa do que se conhecida. Não há conhecimento de relato anterior sobre a utilização desta técnica para determinar contaminantes bacterianos em fermentos de cana-de-açúcar para produção de cachaça

Survey of mycoflora in grains and kernels of maize

Na cultura do milho as podridões de espigas causadas por fungos destacam-se como uma das principais responsáveis pelas perdas em produção e qualidade, principalmente devido a formação dos chamados grãos ardidos. O objetivo deste trabalho foi o de identificar a micoflora presente em grãos e sementes produzidos em difer entes regiões e zonas macro-climáticas do Brasil, nos períodos de safra e safrinha. A determinação da incidência dos gêneros fúngicos foi feita pelo método do papel de filtro com congelamento. Foram a nalisa dos duzentos grãos/ sementes por amostra em um total de 44 amostras de grãos ardidos e 12 de sementes. As análises foram efetuadas sob mi croscópio estereoscópico e microscópio ópt ico. Os principais fungos encontrados neste levantamento, tanto em sementes como em grãos ardidos, foram Penicillum sp., Fusarium spp., Aspergillus spp., Cladosporium sp., Cephalosporium sp. e Stenocarpella spp. Dentre os patógenos principais, Fusarium spp. e Penicillium sp. foram os gêneros encontrados em maior incidência tanto nos grãos como nas sementes nas duas safras. Não houve diferenças significativas entre os diferentes climas e regiões na safra verão para a incidência de Fusarium spp. e Penicillium sp. Porém, durante a safrinha, foi observada uma maior incidência de Fusarium spp. na região CO. Cladosporium sp. se destacou entre os fungos sobretudo durante a safrinha.In maize crop, ear rot caused by fungi is one of the main responsible for quality and production losses, mainly because of the development of the so called discolored grains. The purpose of this study was to identify the mycoflora present in grains and kernels produced in different regions and climatic zones of Brazil during summer and winter harvest and relate the occurrence of fungi with such parameters. The identification of the fungi was done by the filter paper with freezing method. Two hundred grains/kernels for sample totaling 44 samples of discolored grains and 12 of kernels were analyzed. The tests were performed under a stereomicroscope and an optical microscope. The main fungi found in this survey, both in kernel and discolored grains were Penicillium sp., Fusarium spp., Aspergillus spp., Cladosporium sp. Cephalosporium sp. and Stenocarpella spp. Among the major pathogens, Fusarium spp. and Penicillium spp. were found in higher incidence in grains and kernels in both seasons. There were no significant differences between regions and climates in the summer season for Fusarium spp. and Penicillium sp. incidence. However, during the winter season there has been observed a higher incidence of Fusarium spp. in the region CO. Cladosporium sp. stood out among fungi especially during the winter season

Quebra da resistência em pimentão contra o Pepper yellow mosaic virus

Plants of Capsicum annuum cv. Magali R, resistant to Pepper yellow mosaic virus (PepYMV), which showed severe yellow mosaic, leaf malformation and stunting were observed during the 2003/04 growing season in Lins, São Paulo State, Brazil. Potyvirus-like particles observed in leaf sap from infected plants under the electron microscope reacted with an antiserum against PepYMV in PTA-ELISA. In addition to C. annuum cv. Magali R, this potyvirus also infected systemically the resistant C. annuum cv. Rubia R. The nucleotide sequence of part of the CP gene of this potyvirus shared 96-98% identity with that of other PepYMV isolates. The partial nucleotide sequence of the 3' NTR showed 94-96% identity with that of PepYMV. These data indicate that this potyvirus is a resistance-breaking isolate of PepYMV.Plantas de Capsicum annuum cv. Magali R, resistentes ao Pepper yellow mosaic virus (PepYMV), exibindo sintomas severos de mosaico amarelo, malformação foliar e subdesenvolvimento foram encontradas em plantios na região de Lins, SP, Brasil, em 2003/04. Partículas semelhantes àquelas do gênero Potyvirus foram observadas em extrato foliar de planta infectada examinado em microscópio eletrônico de transmissão. O extrato foliar também reagiu com anti-soro contra o PepYMV em PTA-ELISA. Além de C. annuum cv. Magali R, esse potyvirus também infectou sistemicamente C. annuum cv. Rubia R, que é resistente ao PepYMV. A seqüência de nucleotídeos de parte do gene da proteína capsidial (CP) desse potyvirus apresentou 96-98% de identidade com a de outros isolados do PepYMV. A seqüência parcial de nucleotídeos da região 3' não traduzida (3' NTR) apresentou 94-96% de identidade com a do PepYMV. Esses resultados são indicativos de que o potyvirus que quebrou a resistência em pimentão é um isolado do PepYMV

Linkage and mapping of quantitative trait loci associated with angular leaf spot and powdery mildew resistance in common beans

Angular leaf spot (ALS) and powdery mildew (PWM) are two important fungi diseases causing significant yield losses in common beans. In this study, a new genetic linkage map was constructed using single sequence repeats (SSRs) and single nucleotide polymorphisms (SNPs), in a segregating population derived from the AND 277 x SEA 5 cross, with 105 recombinant inbred lines. Phenotypic evaluations were performed in the greenhouse to identify quantitative trait loci (QTLs) associated with resistance by means of the composite interval mapping analysis. Four QTLs were identified for ALS resistance. The QTL ALS11AS, linked on the SNP BAR 5054, mapped on chromosome Pv11, showed the greatest effect (R2 = 26.5%) on ALS phenotypic variance. For PWM resistance, two QTLs were detected, PWM2AS and PWM11AS, on Pv2 and Pv11, explaining 7% and 66% of the phenotypic variation, respectively. Both QTLs on Pv11 were mapped on the same genomic region, suggesting that it is a pleiotropic region. The present study resulted in the identification of new markers closely linked to ALS and PWM QTLs, which can be used for marker-assisted selection, fine mapping and positional cloning

Nosocomial Bloodstream Infections in Brazilian Pediatric Patients: Microbiology, Epidemiology, and Clinical Features

Background: Nosocomial bloodstream infections (nBSIs) are an important cause of morbidity and mortality and are the most frequent type of nosocomial infection in pediatric patients.Methods: We identified the predominant pathogens and antimicrobial susceptibilities of nosocomial bloodstream isolates in pediatric patients (<= 16 years of age) in the Brazilian Prospective Surveillance for nBSIs at 16 hospitals from 12 June 2007 to 31 March 2010 (Br SCOPE project).Results: in our study a total of 2,563 cases of nBSI were reported by hospitals participating in the Br SCOPE project. Among these, 342 clinically significant episodes of BSI were identified in pediatric patients (<= 16 years of age). Ninety-six percent of BSIs were monomicrobial. Gram-negative organisms caused 49.0% of these BSIs, Gram-positive organisms caused 42.6%, and fungi caused 8.4%. the most common pathogens were Coagulase-negative staphylococci (CoNS) (21.3%), Klebsiella spp. (15.7%), Staphylococcus aureus (10.6%), and Acinetobacter spp. (9.2%). the crude mortality was 21.6% (74 of 342). Forty-five percent of nBSIs occurred in a pediatric or neonatal intensive-care unit (ICU). the most frequent underlying conditions were malignancy, in 95 patients (27.8%). Among the potential factors predisposing patients to BSI, central venous catheters were the most frequent (66.4%). Methicillin resistance was detected in 37 S. aureus isolates (27.1%). of the Klebsiella spp. isolates, 43.2% were resistant to ceftriaxone. of the Acinetobacter spp. and Pseudomonas aeruginosa isolates, 42.9% and 21.4%, respectively, were resistant to imipenem.Conclusions: in our multicenter study, we found a high mortality and a large proportion of gram-negative bacilli with elevated levels of resistance in pediatric patients.Pfizer, Inc.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Inst Oncol Pediat IOP GRAAC, São Paulo, BrazilHosp Israelita Albert Einstein, São Paulo, BrazilUniversidade Federal de São Paulo UNIFESP, São Paulo, BrazilHosp 9 Julho, São Paulo, BrazilSanta Casa Porto Alegre, Porto Alegre, RS, BrazilHosp Conceicao, Porto Alegre, RS, BrazilHosp Base, Brasilia, DF, BrazilHosp Walter Cantidio, Fortaleza, Ceara, BrazilHosp Diadema, São Paulo, BrazilHosp Espanhol, Salvador, BA, BrazilHosp Coracao, Natal, RN, BrazilHosp UNIMED, Natal, RN, BrazilHosp Clin Goiania, Goiania, Go, BrazilHosp Rim & Hipertensao, São Paulo, BrazilUniv Fed Triangulo Mineiro, Uberaba, MG, BrazilVirginia Commonwealth Univ, Richmond, VA USAUniversidade Federal de São Paulo UNIFESP, São Paulo, BrazilFAPESP: 2006/57700-0Web of Scienc

Mapping of resistance genes to races 1, 3 and 5 of Podosphaera xanthii in melon PI 414723

The fungus Podosphaera xanthii affects melon crops and presents several races controlled by race-specific resistance genes.The accession PI 414723 is resistant to races 1, 3 and 5 and it is a suitable source of resistance genes. The inheritance of resistanceto these races was analyzed on 87 F2 plants from the cross of PI 414723 × Védrantais, and resistance to all three races could be explainedby the segregation of a single dominant gene, although a digenic model could also be accepted. A genetic map was assembledwith 206 markers, and co-segregation analysis of resistance phenotypes indicated the existence of two linked loci in linkage group II,one conferring resistance to races 1 and 5 (denominated Pm-x1,5), and the second to race 3 (denominated Pm-x3), located 5.1 cMapart. This study reports for the first time the existence of Pm-x3 and the genetic locations of these resistance genes from PI 414723