Alcanivorax dieselolei, an alkane-degrading bacterium associated with the mucus of the zoanthid Palythoa caribaeorum (Cnidaria, Anthozoa) / Alcanivorax dieselolei, uma bactéria degradadora de alcanos associada ao muco do zoantídeo Palythoa caribaeorum (Cnidaria, Anthozoa)

Analyses of 16S rDNA genes were used to identify the microbiota isolated from the mucus of the zoanthid Palythoa caribaeorum at Porto de Galinhas on the coast of Pernambuco State, Brazil. This study is important as the first report of this association, because of the potential biotechnological applications of the bacterium Alcanivorax dieselolei, and as evidence for the presence of a hydrocarbon degrading bacterium in a reef ecosystem such as Porto de Galinhas.

Análises dos genes 16S rDNA foram empregadas para identificar a microbiota isolada do muco do zoantídeo Palythoa caribaeorum de Porto de Galinhas, litoral do estado de Pernambuco, Brasil. Este estudo é importante pelo ineditismo dessa associação, pelas relevantes aplicações biotecnológicas da bactéria Alcanivorax dieselolei e pela indicação da presença de uma bactéria degradadora de hidrocarbonetos em um ecossistema recifal como o de Porto de Galinhas.

Fate of non O157 Shigatoxigenic Escherichia coli in ovine manure composting / Eliminação de Escherichia coli Shigatoxigênica não O157 em compostagem de esterco ovino

Livestock manure may contain pathogenic microorganisms which pose a risk to the health of animal or humans if the manure is not adequately treated or disposed of. To determine the fate of Shiga toxigenic Escherichia coli (STEC) non O157 in composted manure from naturally colonized sheep, fresh manure was obtained from animals carrying bacterial cells with stx1/ stx2 genes. Two composting systems were used, aerated and non-aerated, and the experiments were done in Dracena city, São Paulo State. Every week, for seven weeks, one manure sample from six different points in both systems was collected and cultured to determine the presence of E. coli, the presence of the virulence genes in the cells, and also the susceptibility to 10 antimicrobial drugs. The temperature was verified at each sampling. STEC non-O157 survived for 49 days in both composting systems. E. coli non-STEC showing a high degree of antibiotic resistance was recovered all long the composting period. No relationship was established between the presence of virulence genes and antibiotic resistance. The presence of virulence genes and multiple antibiotic resistances in E. coli implicates a potential risk for these genes spread in the human food chain, which is a reason for concern...

Esterco de animais de criação pode conter microrganismos patogênicos, o que representa um risco para a saúde animal e a humana se o esterco não for adequadamente tratado ou descartado. Determinou-se o tempo necessário para a eliminação de Escherichia coli Shiga toxigenica (STEC) não O157 em esterco ovino composto, obtido de fezes frescas de ovelhas naturalmente colonizadas com cepas STEC não O157 que apresentavam os genes stx1/ stx2. Foram utilizados dois sistemas de compostagem, aerado e não aerado, em experimentos realizados na cidade de Dracena, estado de São Paulo. Todas as semanas, durante sete semanas, uma amostra de compostagem proveniente de seis pontos diferentes na leira, nos dois sistemas, foi coletada e semeada para a determinação da presença de E. coli, da presença de genes de virulência nas células, bem como da sensibilidade dessas células a 10 drogas antimicrobianas. Em cada amostragem, a temperatura da leira foi analisada. Células de STEC não O157 sobreviveram por 49 dias nos dois sistemas de compostagem. E. coli não STEC com um alto grau de resistência a antibióticos foi recuperada ao longo de todo o período de compostagem. Não foi possível estabelecer relação entre a presença de genes de virulência e a resistência a antibióticos. A presença de genes de virulência e a resistência a múltiplos antibióticos em E. coli representam um risco potencial para o espalhamento desses genes na cadeia alimentar humana, o que é motivo de grande preocupação...

Comparação do isolamento microbiológico e da reação em cadeia da polimerase no diagnóstico de salmonelose em bezerros infectados experimentalmente com Salmonella Typhimurium / Comparison of microbiological culture and polymerase chain reaction in the diagnosis of salmonellosis in Salmonella Typhimurium-infected calves

The efficiency of microbiological culture and polymerase chain reaction (PCR) for detection of Salmonella Typhimurium is compared in fecal samples of Holstein calves experimentally infected with 10(9) CFU of Salmonella Typhimurium. Seventy-two fecal samples were analyzed by microbiological culture and PCR associated with selenite cystine (SC) and Muller-Kauffmann tethrationate (TMK) selective enrichment broths. Regardless of the selective enrichment broth, the microbiological culture was significantly better than PCR for detection of positive samples of Salmonella Typhimurium. The selective enrichment broths SC and TMK had no effect on the efficiency of the microbiological culture. The SC broth was the best option as selective enrichment associated to PCR.

Resistência aos antimicrobianos e análise da diversidade genética de Staphylococcus aureus por PCR-RAPD / Resistance to antimicrobials and analysis of Staphylococcus aureus genetic diversity using RAPD-PCR

O objetivo deste trabalho foi analisar a sensibilidade antimicrobiana in vitro de cepas de Staphylococcus aureusisoladas de tetos de vacas e mãos de retireiros, além de verificar o polimorfismo entre elas pela técnica de PCR-RAPD. Os testes foram realizados pela técnica de difusão em discos e, após a extração do material genético foram desenvolvidas as técnicas de PCR e RAPD, usando para isso 40 iniciadores diferentes. A análise do polimorfismo foi realizada empregando-se o programa de taxonomia numérica NTSYS. As sensibilidades dos antimicrobianos nas cepas obtidas de tetos de vacas foram 4% para a penicilina, 88% para a tetraciclina, 92% para a gentamicina, 96% para a vancomicina e 100% ao cloranfenicol. Para as cepas provenientes das mãos de retireiros, os resultados de sensibilidade foram zero para a penicilina, 70% para a tetraciclina e 90% para a vancomicina e 100% para os antimicrobianos gentamicina e cloranfenicol. A realização do E-teste indicou uma concentração inibitória mínima (CIM) maior que 256 mg/mL para as cepas resistentes ao antimicrobiano vancomicina. Os estudos permitiram detectar a resistência dos S. aureus mediante o uso dos antimicrobianos testados e determinar a diversidade genética entre as cepas de estafilococos devido à presença de muitas bandas polimórficas encontradas em todos os iniciadores.

The aim of this study was to analyze in vitro antimicrobial susceptibility of Staphylococcus aureus strains isolated from teats of cow udders and milking workers' hands as well as to verify polymorphism among them by using RAPD-PCR technique. Tests were conducted by disk diffusion technique and after the collection of the genetic material PCR and RAPD techniques were performed with the use of 40 different initiators. The analysis of polymorphism was conducted by using the NTSYS program of numerical taxonomy. The susceptibility of antimicrobials in the strains collected from teats of cow udders was 4% to penicillin, 88% to tetracycline, 92% to gentamicine, 96% to vancomycin and 100% to chloranfenicol. As for the strains collected from milking workers' hands, susceptibility results were 0% to penicillin, 70% to tetracycline and 90% to vancomycin and 100% to gentamicine and chloranfenicol antimicrobials. E-test showed minimum inhibitory concentration (MIC) greater than 256 ?g/mL to strains resistant to the antimicrobial vancomycin. The studies made it possible to detect S. aureus resistance upon the use of the tested antimicrobials and to determine the genetic diversity found among strains of staphylococcus due to the presence of many polymorphic bands found in all initiators.

Avaliação da reação em cadeia da polimerase e do isolamento bacteriológico convencional na detecção de Salmonella Dublin em amostras de fezes de bezerros infectados experimentalmente / Evaluation of polymerase chain reaction and standard microbiological techniques for detection of Salmonella Dublin in fecal samples of infected calves

The efficiency of the Polymerase Chain Reaction (PCR) combined with selective enrichment broth was compared with the standard microbiological techniques for detection of Salmonella Dublin in fecal samples of 10 to 15-days-old Holstein calves, experimentally infected with 10(8) CFU of Salmonella Dublin. Seventy-six fecal samples were analyzed using PCR associated with selenite cystine (SC) and Muller-Kauffmann tetrathionate (TMK) broths and standard microbiological techniques. Regardless of the selective enrichment broth used, the standard microbiological techniques were significantly better than PCR in detection of positive samples of Salmonella Dublin. However, the simultaneous use of both techniques provided detection of a larger number of positive samples. The SC broth was the best option as selective enrichment in both techniques.

The contribution of genes required for anaerobic respiration to the virulence of Salmonella enterica serovar Gallinarum for chickens

Salmonella enterica serovar Gallinarum (SG) is an intracellular pathogen of chickens. To survive, to invade and to multiply in the intestinal tract and intracellularly it depends on its ability to produce energy in anaerobic conditions. The fumarate reductase (frdABCD), dimethyl sulfoxide (DMSO)-trimethylamine N-oxide (TMAO) reductase (dmsABC), and nitrate reductase (narGHIJ) operons in Salmonella Typhimurium (STM) encode enzymes involved in anaerobic respiration to the electron acceptors fumarate, DMSO, TMAO, and nitrate, respectively. They are regulated in response to nitrate and oxygen availability and changes in cell growth rate. In this study mortality rates of chickens challenged with mutants of Salmonella Gallinarum, which were defective in utilising anaerobic electron acceptors, were assessed in comparison to group of bird challenged with wild strain. The greatest degree of attenuation was observed with mutations affecting nitrate reductase (napA, narG) with additional attenuations induced by a mutation affecting fumarate reductase (frdA) and a double mutant (dmsA torC) affecting DMSO and TMAO reductase.

Putative pyrophosphate phosphofructose 1-kinase genes identified in sugar cane may be getting energy from pyrophosphate

Pyrophosphate-dependent phosphofructokinase (PPi-PFK) has been detected in several types of plant cells, but the gene has not been reported in sugar cane. Using Citrus paradisi PPi-PFK gene (AF095520 and AF095521) sequences to search the sugar cane EST database, we have identified both the alpha and beta subunits of this enzyme. The deduced amino acid sequences showed 76 and 80 similarity with the corresponding alpha and beta subunits of C. paradisi. A high degree of similarity was also observed among the PFK b subunits when the alignment of the sugar cane sequences was compared to those of Ricinus communis and Solanum tuberosum. It appears that alpha and beta are two distinct subunits; they were found at different concentrations in several sugar cane tissues. It remains to be determined if the different gene expression levels have some physiological importance and how they affect sucrose synthesis, export, and storage in vacuoles. A comparison between the amino acid sequences of b PFKs from a variety of organisms allowed us to identify the two critical Asp residues typical of this enzyme's activity site and the other binding sites; these residues are tightly conserved in all members of this protein family. Apparently, there are catalytic residues on the b subunit of the pyrophosphate-dependent enzyme

Identification of new isolates of Bacillus thuringiensis using rep-PCR products and d-endotoxin electron microscopy

PCR has been used to analyze the distribution of REP (Repetitive Extragenic Palindromic) and ERIC (Enterobacterial Repetitive Intergenic Consensus) sequences (rep-PCR) found within the genome of the bacterium Bacillus thuringiensis, with the purpose to analyze the genetic similarities among 56 subspecies samples and 95 field isolates. The PCR products were analyzed by EB-AGE (ethidium bromide-agarose electrophoresis) and then submitted to banding comparisons, based on the Phyllip software algorithm. When the banding similarities were considered for comparison purposes among all the strains, the phylogenic tree patterns varied according to the rep-PCR primers considered, but, from a broader point of view, the ERIC sequences produced better results, which, together with electron microscopy analysis of the released parasporal bodies and colony morphology characteristics, allowed to detect two possible new subspecies of B. thuringiensis

A search for markers of sugarcane evolution

Com o propósito de determinar a relaçäo filogenética entre a cana-de-açúcar e membros da subtribo Saccharinae,a regiäo gênica nuclear ITS1-5,8S-ITS2 (ITS: espaçador interno transcrito; 5,8S: DNA ribossomal 5.8S), com alta taxa evolutiva, foi identificada no banco de dados do projeto genoma "Sugarcane Expressed Sequence Tag" (SUCEST). Uma análise através do método de parcimônia, utilizando esta regiäo e seqüências homólogas de 23 Andropogoneae retiradas da base de dados GenBank, indicou que a cana-de-açúcar é o grupo-irmäo de Saccharum sinense. No entanto, devido à pequena quantidade de caracteres informativos para parcimônia e à homoplasia presentes na regiäo ITS1-5,8S-ITS2, näo foi possível determinar com segurança a relaçäo filogenética entre a cana-de-açúcar e alguns dos demais membros da tribo Saccharine. Como alternativa para esta baixa resoluçäo, dezessete regiões gênicas nucleares, cloroplasmáticas ou mitocondriais foram selecionadas a partir do banco de dados SUCEST com o objetivo de encontrar marcadores mais apropriados para a reconstruçäo da filogenia da cana-de-açúcar. Entre elas, aquelas correspondentes à alfa-tubulina, rpl16, a rpoC2 apresentaram baixa incidência de polimorfismo e taxas de evoluçäo equivalentes ou mesmo maiores do que a observada para a regiäo ITS1-5,8S-ITS2. Estes marcadores säo propostos como preferenciais para estudos filogenéticos da subtribo Saccharinae.