Light and scanning electron microscopy of Henneguya arapaima n. sp (Myxozoa: Myxobolidae) and histology of infected sites in pirarucu (Arapaima gigas: Pisces: Arapaimidae) from the Araguaia River, Brazil

In this report, we describe Henneguya arapaima n. sp., a parasite of the gill arch and gall bladder of Arapaima gigas (pirarucu) collected in the Araguaia River, in the municipality of Nova Crixas, Goias State, central Brazil. The plasmodia were white, round or ellipsoidal and measured 200-600 mu m. Parasite development was asynchronous and the mature spores were fusifonn and had smooth wall. The spores measurements were (range, with means +/- S.D. in parentheses): total length-48.4-53.1 mu m (51.6 +/- 3.4 mu m), body length-13.5-15.2 mu m (14.2 +/- 0.8 mu m), body width-5.1-6.1 mu m (5.7 +/- 0.5 mu m), body thickness-4.7-5.3 mu m (4.9 +/- 0.2 mu m) and caudal process length-38.0-41.2 mu m (38.3 +/- 2.9 mu m). The polar capsules were elongated and of unequal size, with lengths of 6.3-6.8 mu m (6.5 +/- 0.2) and 6.2-6.6 mu m (6.3 +/- 0.1) for the longest and shortest axes, respectively. Capsule width was 1.4-1.6 mu m (1.5 +/- 0.1). Histological analysis showed that the plasmodia occurred in the tunica adventitia of the gall bladder and were delimited by a thin capsule of connective tissue. In the gill arch, the plasmodia were also surrounded by connective tissue similar to the endomesium, of striated skeletal muscle cells. Sixty-five juvenile specimens of A. gigas weighing 1.0-25.0 kg were examined, 17 (26.1%) of which were infected. Of these, 14 (82.3%) had cysts in the gall bladder, two (11.7%) had cysts in the gill arch and only one (5.9%) had cysts in both organs. When the fish were grouped by weight, the prevalence of infection in fish weighing up to 10.0 kg (20.7%) was significantly lower than in fish weighing 10.1-25.0 kg (50%) (G = 3.93; d.f. = 1; p < 0.05). (C) 2008 Elsevier B.V. All rights reserved.

Study of the relationship between roads, relief, land use, and natural vegetation in the Ibiuna Plateau - SP, focusing on landscape ecology

Roads facilitate access by deforestation agents, being relevant in studies approaching conservationist matters in rainforests. It is important to understand the relationship between road distribution, relief, land use, and forest coverage in order to evaluate where forests are more vulnerable. This study aimed at: 1) understanding the relationship between relief and density and road connectivity in three moments in time; and 2) evaluating the relationship between distance from roads and forest coverage, farmlands and rural and urban facilities in a fragmented Atlantic Forest landscape in three moments in time. Maps of roads, altitude, and land use and coverage were used. Chi-square tests showed that: 1) density and road connectivity did not present significant relationship with the relief; and 2) forest areas occupy areas distant from the roads, while farmlands and rural and urban facilities occupy areas nearer the roads. Roads and land use, regardless of relief, influence forest coverage distribution. Thus, we suggest that roads are taken into account in conservationist strategies and environmental planning.

Naming the Bonaire banded box jelly, Tamoya ohboya, n. sp. (Cnidaria: Cubozoa: Carybdeida: Tamoyidae)

A new species of cubozoan jellyfish has been discovered in shallow waters of Bonaire, Netherlands ( Dutch Caribbean). Thus far, approximately 50 sightings of the species, known commonly as the Bonaire banded box jelly, are recorded, and three specimens have been collected. Three physical encounters between humans and the species have been reported. Available evidence suggests that a serious sting is inflicted by this medusa. To increase awareness of the scientific disciplines of systematics and taxonomy, the public has been involved in naming this new species. The Bonaire banded box jelly, Tamoya ohboya, n. sp., can be distinguished from its close relatives T. haplonema from Brazil and T. sp. from the southeastern United States by differences in tentacle coloration, cnidome, and mitochondrial gene sequences. Tamoya ohboya n. sp. possesses striking dark brown to reddish-orange banded tentacles, nematocyst warts that densely cover the animal, and a deep stomach. We provide a detailed comparison of nematocyst data from Tamoya ohboya n. sp., T. haplonema from Brazil, and T. sp. from the Gulf of Mexico.

Role of CcpA in Polyhydroxybutyrate Biosynthesis in a Newly Isolated Bacillus sp MA3.3

The ccpA gene was inactivated in the polyhydroxybutyrate (PHB)-producing strain Bacillus sp. MA3.3 in order to reduce glucose catabolite repression over pentoses and develop improved bacterial strains for the production of PHB from lignocellulosic hydrolysates. Mutant Bacillus sp. MSL7 Delta CcpA are unable to grow on glucose and ammonia as sole carbon and nitrogen sources, respectively. Supplementation of glutamate as the nitrogen source or the substitution of the carbon source by xylose allowed the mutant to partially recover its growth performance. RT-PCR showed that CcpA stimulates the expression of the operon (gltAB), responsible for ammonia assimilation via glutamate in Bacillus sp. MA3.3. Moreover, it was demonstrated that the supplementation of xylose or glutamate was capable of stimulating gltAB operon expression independently of CcpA. In PHB production experiments in mineral media, it has been observed that the glucose catabolite repression over the pentoses was partially released in MSL7. Although the carbohydrate consumption is faster in the ccpA mutant, the biomass and PHB biosynthesis are lower, even with supplementation of glutamate. This is attributed to an increase of acetyl-CoA flux towards the tricarboxylic acid cycle observed in the mutant. Copyright (C) 2011 S. Karger AG, Basel

Phylogenetic Analyses Based on Small Subunit rRNA and Glycosomal Glyceraldehyde-3-Phosphate Dehydrogenase Genes and Ultrastructural Characterization of Two Snake Trypanosomes: Trypanosoma serpentis n. sp from Pseudoboa nigra and Trypanosoma cascavelli from Crotalus durissus terrificus

We sequenced the small subunit (SSU) rRNA and glycosomal glyceraldehyde-3-phosphate dehydrogenase (gGAPDH) genes of two trypanosomes isolated from the Brazilian snakes Pseudoboa nigra and Crotalus durissus terrificus. Trypanosomes were cultured and their morphometrical and ultrastructural features were characterized by light microscopy and scanning and transmission electron microscopy. Phylogenetic trees inferred using independent or combined SSU rRNA and gGAPDH data sets always clustered the snake trypanosomes together in a clade closest to lizard trypanosomes, forming a strongly supported monophyletic assemblage (i.e. lizard-snake clade). The positioning in the phylogenetic trees and the barcoding based on the variable V7-V8 region of the SSU rRNA, which showed high sequence divergences, allowed us to classify the isolates from distinct snake species as separate species. The isolate from P. nigra is described as a new species, Trypanosoma serpentis n. sp., whereas the isolate from C. d. terrificus is redescribed here as Trypanosoma cascavelli.

Genotyping, physiological features and proteolytic activities of a potentially pathogenic Acanthamoeba sp isolated from tap water in Brazil

Acanthamoeba spp., known to cause keratitis and granulomatous encephalitis in humans, are frequently isolated from a variety of water sources. Here we report for the first time the characterization of an Acanthamoeba sp. (ACC01) isolated from tap water in Brazil. This organism is currently being maintained in an axenic growth medium. Phylogenetic analysis based on SSU rRNA gene sequences positioned the new isolate in genotype T4, closest to the keratitis-causing isolate, A. polyphaga ATCC 30461 (similar to 99% similarity). Acanthamoeba ACC01 and A. polyphaga 30461 both grew at 37 degrees C and were osmotically resistant, multiplying in hyperosmolar medium. Both isolates secreted comparable amounts of proteolytic enzymes, including serine peptidases that were optimally active at a near neutral/alkaline pH and resolved identically in gelatin gels. Incubation of gels at pH 4.0 with 2 mM DTT also indicated the secretion of similar cysteine peptidases. Altogether, the results point to the pathogenic potential of Acanthamoeba ACC01. (C) 2009 Elsevier Inc. All rights reserved.

Geochemistry of iron and manganese in soils and sediments of a mangrove system, Island of Pai Matos (Cananeia - SP, Brazil)

Five zones along a transect of 180 m were selected for study on the Island of Pai Matos (Sao Paulo, Brazil). Four of the zones are colonised by vascular plants (Spartina SP, Laguncularia LG, Avicennia AV and Rhizophora RH) and were denominated soils, and the other zone, which lacks vegetation, was denominated sediment (SD). The geochemical conditions differed significantly in soils and sediment and also at different depths. The soils were oxic (Eh > 350 mV) or suboxic (Eh: 350-100 mV) at the surface and anoxic (Eh < 100 mV) at depth, whereas in the sediment anoxic conditions prevailed at all depths, but with a lower concentration of sulphides in the pore water and pyrite in the solid fraction. Under these geochemical conditions Fe is retained in the soils, while the Mn tends to be mobilized and lost. The most abundant form of iron oxyhydroxide was lepidocrocite (mean concentration for all sites and depths, 45 +/- 19 mu mol g(-1)), followed by goethite (30 19 mu mol g(-1))and ferrihydrite (19 +/- 11 mu mol g(-1)),with significant differences among the mean concentrations. There was a significant decrease with depth in all the types of Fe oxyhydroxides measured, particularly the poorly crystalline forms. The pyrite fraction was an important component of the free Fe pool (non-silicate Fe) in all soils as well as in the sediment, especially below 20 cm depth (mean concentration for all sites and depths, 60 +/- 54 mu mol CI). Furthermore, the mean concentration of Fe-pyrite for all sites and depths was higher than that obtained for any of the three Fe oxyhydroxides measured. The Fe-AVS was a minor fraction, indicating that the high concentrations of dissolved Fe in the soils in the upper area of the transect result from the oxidation of Fe sulphides during low tide. Mossbauer spectroscopy also revealed that most of the Fe (III) was associated with silicates, in this case nontronite. The presence of crystals of pyrite associated with phyllosilicates in samples from the upper layer of the soils may indicate that pyritization of this form of Fe(III) is more rapid than usually reported for ocean bed sediments. The sequential extraction of Mn did not reveal any clearly dominant fraction, with the Mn-carbonate fraction being the most prevalent, followed by exchangeable Mn and oxides of Mn, whereas pyrite-Mn and Mn associated with crystalline Fe-oxides were present at significantly lower concentrations. The high concentration of dissolved Mn found in the soils in the lower part of the transect is consistent with the fact that the solubility is determined by the carbonate fraction. Unlike for Fe, in the soils in the higher zone, which are subject to intense drainage during low tide, there was loss of Mn, as reflected by the concentration of total Mn. (C) 2008 Elsevier B.V. All rights reserved.

CHRONOLOGICAL DISTRIBUTION OF BRAZILIAN GLYPTODON SP REMAINS: A DIRECT (14)C DATE FOR A SPECIMEN FROM IPORANGA, SAO PAULO, BRAZIL

Glyptodon sp. fossil remains can be found throughout Brazil. However, little information is available about their chronological distribution. With the intention to contribute to this issue, we present, as far as we know, the first direct radiocarbon date for 1 specimen of this genus found in Brazil. The osteoderm MZSP-PV660 found in Abismo do Fossil Cave (SP-145), Iporanga, Sao Paulo, Brazil, was dated by accelerator mass spectrometry at the Beta Analytic Radiocarbon Dating Laboratory. The (14)C date obtained was between 20,680 and 21,370 calibrated years before the present. Unfortunately, the scant (and often imprecise or unreliable) chronological data regarding this species and genus in Brazil and elsewhere in South America precludes a robust comparison among the dates available and the one presented here. Nevertheless, our finding supports the existence of this genus in South America at least until the Last Glacial Maximum.

Sesquiterpenes from Xylaria sp., an endophytic fungus associated with Piper aduncum (Piperaceae)

Two new presilphiperfolane sesquiterpenes, 1 and 2, were isolated from the ethyl acetate extract of Xylaria sp., obtained from the leaves of Piper aduncum, along with two known eremophilane sesquiterpenes, phaseolinone (3) and phomenone (4). Chemical structures of 1 and 2 were established by analysis of spectroscopic data. The four compounds were tested in vitro for antifungal and cytotoxicity activities using CHO (Chinese hamster ovary). Compounds 1 and 2 did not show any antifungal and cytotoxic activity. Compounds 3 and 4 displayed moderate cytotoxic activities, as well as 4 antifungal activity. (C) 2010 Phytochemical Society of Europe. Published by Elsevier B. V. All rights reserved.

Produção, caracterização, purificação e aplicação de uma protease produzida pelo microrganismo Microbacterium sp. kr10

O uso de enzimas como agentes de modificação das propriedades funcionais de proteínas tem se tornado bastante difundido na indústria de alimentos. As proteases, apresentam inúmeras vantagens, principalmente, devido a sua atividade em baixas concentrações e a sua ausência de toxicidade, que faz com que se elimine a necessidade da sua remoção do produto final. O objetivo deste trabalho foi determinar as condições ótimas de produção da protease de Microbacterium sp. kr10, caracterizar e purificar parcialmente a enzima, assim como verificar a sua utilização como agente de modificação das propriedades funcionais da proteína de soja. Através da metodologia de superfície de resposta foram determinadas as condições ótimas de produção da protease, pH de 7,0, temperatura de 25°C e 12,5 g L-1 de farinha de pena (p/v). O padrão proteolítico da enzima tanto no extrato cru quanto na parcialmente purificada indicam que esta é uma metaloprotease, com pH e temperaturas ótimos nas faixas de 6,5 a 7,5, e 45 a 55°C, respectivamente. A atividade enzimática foi totalmente inibida por EDTA, fenantrolina, HgCl2 e CuCl2 e parcialmente inibida por ZnCl2, MnCl2 e SnCl2. A enzima foi parcialmente purificada através de cromatografia de gel filtração e troca iônica resultando num fator de purificação de 250. Um aumento gradativo do grau de hidrólise da proteína de soja foi observado à medida que se aumentou a razão enzima/substrato utilizada, assim como a redução da formação de espuma e o aumento da capacidade emulsificante de uma solução composta pelo hidrolisado de soja e óleo de soja, mesmo sob condições de alta temperatura e alta concentração de sal. Desta forma, esta protease apresenta potencial para aplicação como agente de modificação protéica de proteína de soja isolada.

Comparação de dois métodos de quantificação de Salmonella sp. em embutidos suínos.

A segurança dos alimentos é uma preocupação mundial e um fator importante na comercialização de produtos de origem animal. A presença de Salmonella sp. em suínos ao abate e em produtos do tipo frescal podem representar um risco para a saúde do consumidor. A análise de risco prevê a avaliação de diferentes fatores, entre eles a quantificação do microrganismo presente no alimento. A partir disso, a contribuição do presente estudo foi buscar estabelecer um método confiável de quantificação de Salmonella sp. em produtos suínos, uma vez que uma das etapas da análise de risco prevê a quantificação do perigo. No caso de Salmonella sp., a técnica da estimativa do Número Mais Provável (NMP) tem sido adotada. Em uma primeira fase desse trabalho, amostras foram quantificadas, individualmente, com três amostras de Salmonella Typhimurium (ATTCC15290 e 2 amostras de suínos) em três diferentes contagens, 101, 102 e 103 UFC. Para o método A, as amostras quantificadas foram adicionadas a 225 mL de água peptonada tamponada, sendo, posteriormente, fracionadas em 3 alíquotas de 50mL, 5mL e 0,5mL. Para o método B, a amostra fortificada foi diluída em água peptonada tamponada até 10-3, sempre em triplicata. Na segunda fase, foram testadas amostras naturalmente contaminadas, utilizando as mesmas metodologias usadas na primeira fase. Todas as alíquotas de ambos métodos foram incubadas a 370C por 18 horas. Após, cada alíquota foi semeada em caldo Rappaport-Vassiliadis (RV) e incubadas à 420C por 24 h e após, em àgar Xylose-Lysine-Tergitol 4 (XLT4) à 370C por 48 h. Colônias suspeitas foram confirmadas por testes bioquímicos. O número de placas positivas para Salmonella sp. foi utilizado para o cálculo do Número Mais Provável, utilizando tabela apropriada. Na segunda fase, os dois métodos foram avaliados em 20 amostras naturalmente contaminadas, mantidas congeladas por até 115 dias. Em 45 ensaios conduzidos, para cada método, em amostras de lingüiça de carne suína contaminadas artificialmente, 38 do método A e 41 do método B resultaram em NMP (95% de intervalo de confiança) concordante com número de UFC de Salmonella inoculado. A maioria das amostras naturalmente contaminada de massa de embutido apresentaram contagens <10NMP/g. A variabilidade encontrada entre os valores de NMP médio foi bastante elevada, tanto entre os métodos como entre repetições de um mesmo método. Isto reflete uma das limitações do método de NMP para estimar a contagem de microrganismos e deverá ser considerada quando o método for aplicado em alimentos naturalmente contaminados e quando aplicado em um estudo de análise de risco. Uma vez que o método B foi o que demonstrou valores médios de NMP mais próximos das quantidades inoculadas, sugere-se que este seja adotado em estudos futuros de quantificação de Salmonella sp. em produtos de origem suína.

Isolamento de bactérias resistentes a cromo hexavalente e purificação parcial da enzima redutora de cromo do Bacillus sp. ES29

A redução do Cr(VI) para Cr(III) diminui a toxidade deste metal no ambiente uma vez que, o Cr(III) é insolúvel às membranas biológicas. Assim a redução microbiana do Cr(VI) é uma alternativa para reduzir os impactos ambientais causados por este metal, utilizado em diversos processos industriais. O objetivo deste trabalho foi selecionar microrganismos a partir de solo contaminado com cromo, caracterizar sua capacidade de redução do Cr(VI) durante o crescimento celular e purificar parcialmente a enzima cromo redutase do Bacillus sp. ES29, através da precipitação com sulfato de amônio (45-75%), cromatografia de gel filtração (Sephadex G-25) e cromatografia de interação hidrofóbica (Octyl Sepharose). A atividade de redução do Cr(VI) pelos isolados foi quantificada com o reagente de s-difenilcarbazida. No isolamento, foram obtidas 20 bactérias resistentes a cromo(VI). Seis destas foram capazes de reduzir 100 mg L-1 Cr(VI) em 24 horas. Um dos isolados foi identificado, através de testes bioquímicos, como pertencete ao gênero Bacillus, sendo tolerante a 750 mg L-1 Cr(VI) e reduzindo mais de 40% do Cr(VI) durante o crescimento celular. Na purificação parcial da enzima foi obtido um fator de purificação de 11,2, aumentando a atividade específica da enzima acima de 11 vezes, porém se faz necessário mais passos de purificações para obtenção desta enzima pura. As bactérias selecionadas e a enzima parcialmente purificada, foram eficientes na redução do Cr(VI) e apresentam potencial para outros estudos, visando a aplicação em processos de biorremediação.

Purificação e caracterização de uma β-glucuronidase extraída do molusco mesogastropoda ampularidade Pomacea sp

This work studies the involved enzymatic way in the metabolism of glycosaminoglycans sulfateds in the mollusc Pomacea sp. Had been identified endoglycosidases and exoglycosidases in the enzymatic extract of the mollusc Pomacea sp by means of hydrolysis activity in condroitim sulphate of whale cartilage and of the p-Nitrofenil-β-glucuronide, respectively. The enzymatic extracts qere obtained of Pomacea sp. being used of 0.1 sodium acetate buffer, pH 5.0 and later centrifugated the 8,000 x g and the presents proteins in the sobrenadante were submitted to the fractionament with two crescents ammonium sulphate concentrations, the visualized activity biggest in the F2 fraction (50-80%). The β-glucuronidase (F3) was isolated in gel chromatography filtration Biogel 1.5m, the purification degree was ratified in Chromatography Liquid of high efficiency (HPLC). The enzyme was purificated 6.362,5 times with 35,6% yield. The β -glucuronidase isolated in this work showed a molecular mass of 100 kDa, determined for eletroforese in poliacrilamida gel . The determination of the ideal kinetic parameters for the catalysis of the p-nitrofenil- β -glucuronide for β-glucuronidase, showed excellent activity in pH 5,0 and temperature 65ºC for 6 hours and apparent Km of 72 x 10-2 mM. It is necessary for the total degradation of 3mM of p-N-β-glucoronide, the amount of 1,2μg of ss-glucuronidase. The BaCl2 increased the activity of ss-glucuronidase, and the activity was inhibited completely by the composites SDS and NaH2PO4