Fungal and fungal-like diversity present in ornithogenically influenced maritime Antarctic soils assessed using metabarcoding.

We assessed soil fungal and fungal-like diversity using metabarcoding in ornithogenically influenced soils around nests of the bird species Phalacrocorax atriceps, Macronectes giganteus, Pygoscelis antarcticus, and Pygoscelis adelie on the South Shetland Islands, maritime Antarctic. A total of 1,392,784 fungal DNA reads was obtained and assigned to 186 amplicon sequence variants (ASVs). The dominant fungal phylum was Ascomycota, followed by Basidiomycota, Chytridiomycota, Blastocladiomycota, Rozellomycota, Mortierellomycota, Monoblepharomycota, Aphelidiomycota, Basidiobolomycota, Mucoromycota, and the fungal-like Oomycota (Stramenopila), in rank order. Antarctomyces sp., Blastocladiomycota sp., Pseudogymnoascus pannorum, Microascaceae sp., Mortierella sp., Lobulomycetales sp., Sordariomycetes sp., Fungal sp., Rhizophydiales sp., Pseudeurotiaceae sp., Chytridiomycota sp. 1, Filobasidiella sp., Tausonia pullulans, Betamyces sp., and Leucosporidium sp. were the most abundant assigned taxa. The fungal assemblages present in the different ornithogenically influenced soils displayed different diversity indices. However, in general, we detected high fungal diversity and few taxa shared between the samples. Despite the polyextreme environmental conditions experienced in these Antarctic soils, the metabarcoding approach detected a rich and complex fungal community dominated by saprophytes, but with some pathogenic taxa also present. The community was dominated by psychrophilic and psychrotolerant taxa, some apparently endemic to Antarctica, and those identified only at higher taxonomic levels, which may represent currently undescribed fungi. The mycobiome detected included taxa characterized by different ecological roles, including saprotrophic, human- and animal-associated, phytopathogenic, mutualistic, and cosmopolitan. These fungi may potentially be dispersed by birds or in the air column over great distances, including between different regions within Antarctica and from South America, Africa, and Oceania.

Selection of Antarctic yeasts as gray mold biocontrol agents in strawberry.

The postharvest disease popularly known as gray mold is considered one of the most limiting factors strawberry fruit production. The most effective way to control this disease is still the use of chemical fungicides. However, other alternative sources of control are being explored. Among these, psychrophilic yeasts adapted to extreme conditions, such as those found in the Antarctic region, may have great potential for use as biocontrol agents. Thus, the present study aimed to select psychrotolerant yeasts obtained from Antarctic region and to evaluate their potential for biocontrol under gray mold, caused by Botrytis cinerea in strawberries stored at low temperature. For this, 20 potential antagonist yeasts were evaluated in vitro (thermotolerance and enzymatic) assays. Debaryomyces hansenii, Rhodotorula mucilaginosa and Dioszegia hungarica were selected for growing in strawberry juice. However, only D. hansenii was selected for in vivo studies and showed a reduction in the incidence of gray mold by 82% for the tests performed on injury and 86% for the tests on non-injured fruits treated by immersion bath. Thus, demonstrating that the selection of this cold-adapted Antarctic yeast can be a promising strategy as a biocontrol agent used to curb the development of gray mold in strawberry fruits.

In Vitro Cytotoxic Effects and Mechanisms of Action of Eleutherine Isolated from Eleutherine plicata Bulb in Rat Glioma C6 Cells.

Gliomas are the most common primary malignant brain tumors in adults, and have a poor prognosis, despite the different types of treatment available. There is growing demand for new therapies to treat this life-threatening tumor. Quinone derivatives from plants have received increased interest as potential anti-glioma drugs, due to their diverse pharmacologic activities, such as inhibiting cell growth, inflammation, tumor invasion, and promoting tumor regression. Previous studies have demonstrated the anti-glioma activity of Eleutherine plicata, which is related to three main naphthoquinone compounds-eleutherine, isoeleutherine, and eleutherol-but their mechanism of action remains elusive. Thus, the aim of this study was to investigate the mechanism of action of eleutherine on rat C6 glioma. In vitro cytotoxicity was evaluated by MTT assay; morphological changes were evaluated by phase-contrast microscopy. Apoptosis was determined by annexin V-FITC-propidium iodide staining, and antiproliferative effects were assessed by wound migration and colony formation assays. Protein kinase B (AKT/pAKT) expression was measured by western blot, and telomerase reverse transcriptase mRNA was measured by quantitative real-time polymerase chain reaction (qRT-PCR). Eleutherine reduced C6 cell proliferation in a dose-dependent manner, suppressed migration and invasion, induced apoptosis, and reduced AKT phosphorylation and telomerase expression. In summary, our results suggest that eleutherine has potential clinical use in treating glioma.

Taxonomy and richness of yeasts associated with angiosperms, bryophytes, and meltwater biofilms collected in the Antarctic Peninsula.

The interest in the diversity of yeasts in the Antarctic environment has increased in recent years, mainly because Antarctic microbiology is a recent science, and little is known about the biodiversity and genetic resources of the microorganisms that inhabit this ecosystem. This study aimed to determine the diversity of epiphytic yeasts in samples of Deschampsia antarctica, Colobanthus quitensis, and bryophytes, as well yeasts present in biofilms collected from Antarctic meltwater. Samples were collected in the summer of 2014 and 2015 during expeditions organized by the Brazilian Antarctic Program. A total of 310 yeasts were isolated, and 34 species were identified by sequencing the D1/D2 domains of the rDNA region belonging to 18 genera. The species Vishniacozyma victoriae and Mrakia gelida were the most abundant. Dioszegia antarctica and Leucosporidium creatinivorum were found only in plant substrates. Most psychrophilic yeasts were isolated from biofilms, including Glaciozyma antarctica, Glaciozyma martinii, Mrakia gelida, Mrakia frigida, Mrakia robertii, Phenoliferia glacialis, and Phenoliferia psychrophenolica, suggesting that the substrates examined in this study represented an interesting habitat for the isolation and characterization of epiphytic and non-epiphytic yeasts that colonize the Antarctic region.

Phytochemistry and antibacterial activity of aqueous andhydroalcoholic extracts of three medicinal plants against foodpathogens / Fitoquímica e atividade antibacteriana de extratos aquosos e hidroalcoólicos de trêsplantas medicinais contra patógenos de alimentos

This study aimed to characterize the phytochemical compositions of three medicinal Brazilian plants’ leaves and bast extracts, and to determine their antibacterial activity on three foodborne and waterborne bacterial pathogens. Parkia platycephala, Pouteria ramiflora and Lophanthera lactescens leaves and basts were collected and aqueous and hydroalcoholic extracts were prepared. Qualitative screening of the phytochemical extracts was performed with three replicates and in triplicate in order to identify the bioactive compounds. The Minimal Inhibitory Concentration and Minimal Bactericide Concentration were determined by microdilution in broth and Escherichia coli, Salmonella typhimurium and Staphylococcus aureus growth was observed on agar plates. Phytochemical composition analysis allowed for the identification of anthraquinones, catechins, saponins, tannins, sesquiterpenlactones and other lactones in the three plants’ leaves and bast aqueous and hydroalcoholic extracts. Eighty-three percent of the plant extracts showed antibacterial activity against S. aureus, and P. platycephala extracts were the only ones that inhibited E. coli and S. typhimurium growth. The present study contributes significantly to the phytochemical composition characterization of three plant species commonly used in Brazilian traditional medicine. The plant extracts’ in vitro antibacterial activity was demonstrated and catechins present in the extracts are, most likely, the bioactive compounds responsible for this action.

Este estudo objetivou caracterizar a composição fitoquímica dos extratos de folhas e das entrecascas de três plantas medicinais brasileiras e determinar a sua atividade antimicrobiana contra três patógenos bacterianos de alimentos. Foram elaborados extratos aquosos e hidroalcoólicos, por meio de folhas e de entrecascas de Parkiaplatycephala, Pouteriaramiflora e Lophantheralactescens. O estudo qualitativo dos extratos foi realizado com três réplicas, em triplicata, para permitir a identificação dos compostos bioativos. A Concentração Inibitória Mínima e a Concentração Bactericida Mínima foram determinadas por microdiluição contra Escherichia coli, Salmonella typhimurium e Staphylococcus aureus. A análise da composição fitoquímica permitiu identificar antraquinonas, catequinas, saponinas, taninas, sesquiterpenlactonas e outras lactonas nos extratos aquosos e hidroalcoólicos das folhas e das entrecascas das três plantas. Oitenta e três porcento dos extratos das plantas apresentaram atividade antibacteriana contra S. aureus. Os extratos de P. platycephala foram os únicos que inibiram o crescimento de E. coli e S. typhimurium. Este estudo contribui significativamente para a caracterização da composição fitoquímica de três espécies de plantas, frequentemente, utilizadas na medicina tradicional brasileira. A atividade antibacteriana, in vitro, dos extratos das plantas foi demonstrada, e as catequinas são, provavelmente, o composto bioativo responsável por essa atividade.

Production of cellulases by Penicillium oxalicum through solid state fermentation using agroindustrial substrates / Produção de celulases por Penicillium oxalicum em fermentação em estado sólido usando diferentes substratos agroindustriais

The purpose of this study was to define the factors that influence the production of cellulases by Penicillium oxalicum, a strain obtained from a leaf-cutting ant colony and identified based on the ITS gene. The experimental design included solid state fermentation using sugarcane bagasse and lignocellulosic sorghum as the lignocellulosic substrate. The variables were analyzed using a 25-1 fractional factorial design, with three replicates on the central point. All the variables analyzed influenced the production of at least one of the three cellulose types analyzed. The highest values observed were: FPase 4.2 U g-1, CMCase 9.2 U g-1 and Avicelase 8.4 U g-1 using lignocellulosic sorghum as the substrate. The best conditions for enzyme production were: incubation temperature at 40ºC, initial moisture of 60%, pH of 4.0 and a growth period of four days using lignocellulosic sorghum as the substrate.

O estudo teve como foco a determinação de fatores que influenciam a produção de celulases por uma cepa isolada de ninho de formigas cortadeiras e identificada por meio do gene ITS como Penicillium oxalicum. O processo produtivo foi Fermentação em Estado Sólido utilizando como substrato lignocelulósico bagaço de cana-de-açúcar e sorgo lignocelulósico. As variáveis foram analisadas através de um planejamento fatorial fracionário 25-1, com três repetições no ponto central. Todas as variáveis analisadas influenciaram a produção de pelo menos um dos três tipos de celulases analisados. As maiores atividades observadas foram: FPase 4,2 U g-1; CMCases 9,2 U g-1 e avicelase 8,4 U g-1, utilizando sorgo lignocelulósico como substrato. As melhores condições para produção foram: temperatura de incubação a 40oC, umidade inicial 60%, pH 4,0, tempo de cultivo de quatro dias, utilizando como substrato sorgo lignocelulósico.

Chemical Composition and Antimicrobial Potential of Palm Leaf Extracts from Babaçu (Attalea speciosa), Buriti (Mauritia flexuosa), and Macaúba (Acrocomia aculeata).

Babaçu (A. speciosa), Buriti (M. flexuosa), and Macaúba (A. aculeata) are palm trees typical of the ecotone area between Cerrado and the Amazon rainforest. The purpose of this study was to evaluate the antimicrobial potential of the extracts prepared from the leaves of those palms as well as determine their chemical compositions. The ethanol extracts were prepared in a Soxhlet apparatus and tested by disk diffusion and agar dilution technique against Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, Pseudomonas aeruginosa, Candida albicans, and Candida parapsilosis. However, there was no significant activity at concentrations of 25, 50, and 100 mg·Ml(-1). Moreover, the phytochemical analysis revealed the presence of tannins, flavonoids, catechins, steroids, triterpenes, and saponins. Gas chromatography (GC/MS) analysis also identified organic acids, such as capric (decanoic) acid, lauric (dodecanoic) acid, myristic (tetradecanoic) acid, phthalic (1,2-benzenedicarboxylic) acid, palmitic (hexadecanoic) acid, stearic (octadecanoic) acid, linoleic (9,12-octadecadienoic) acid (omega-6), linolenic (octadecatrienoic) acid (omega-3), and the terpenes citronellol and phytol. Based on the chemical composition in the palm leaf extracts, the palms have the potential to be useful in the food, cosmetic, and pharmaceutical industries.

Biological activities of ophiobolin K and 6-epi-ophiobolin K produced by the endophytic fungus Aspergillus calidoustus.

Endophytic fungi represent ubiquitous microbial organisms able to live in the tissues of different plants around the world and represent a prolific source of bioactive metabolites. In the present study, the endophytic fungus Aspergillus calidoustus was isolated from the medicinal plant Acanthospermum australe (Asteraceae), and identified using molecular, physiological and morphological methods. A methylene chloride crude extract of A. calidoustus has been produced and subjected to antifungal bioassay-directed fractionation which resulted in the isolation of the two bioactive compounds: ophiobolin K and 6-epi-ophiobolin K. These pure compounds displayed antifungal activity against fungal plant pathogens, protozoal activity against Trypanosoma cruzi, and cytotoxic activity against human tumoral cell lines. The results show that A. calidoustus was able to produce the antifungal and cytotoxic metabolites ophiobolin K and 6-epi-ophiobolin K, which may help the fungus to colonise and occupy the substratum as well as survive in natural environments.

Use of Probiotics to Control Aflatoxin Production in Peanut Grains.

Probiotic microorganisms (Saccharomyces cerevisiae var. boulardii, S. cerevisiae UFMG 905, and Lactobacillus delbrueckii UFV H2b20) were evaluated as biological control agents to reduce aflatoxin and spore production by Aspergillus parasiticus IMI 242695 in peanut. Suspensions containing the probiotics alone or in combinations were tested by sprinkling on the grains followed by incubation for seven days at 25°C. All probiotic microorganisms, in live and inactivated forms, significantly reduced A. parasiticus sporulation, but the best results were obtained with live cells. The presence of probiotics also altered the color of A. parasiticus colonies but not the spore morphology. Reduction in aflatoxin production of 72.8 and 65.8% was observed for S. boulardii and S. cerevisiae, respectively, when inoculated alone. When inoculated in pairs, all probiotic combinations reduced significantly aflatoxin production, and the best reduction was obtained with S. boulardii plus L. delbrueckii (96.1%) followed by S. boulardii plus S. cerevisiae and L. delbrueckii plus S. cerevisiae (71.1 and 66.7%, resp.). All probiotics remained viable in high numbers on the grains even after 300 days. The results of the present study suggest a different use of probiotics as an alternative treatment to prevent aflatoxin production in peanut grains.

Chemical Assessment and Antimicrobial and Antioxidant Activities of Endophytic Fungi Extracts Isolated from Costus spiralis (Jacq.) Roscoe (Costaceae).

Costus spiralis (Costaceae) is a species native to the Amazon region and is used in traditional medicine. The endophytic fungi used in this study were obtained from leaves of this plant. 13 strains were selected to obtain hydroethanolic extracts and were submitted to hydroalcoholic extraction and evaluated for antioxidant activity by DPPH (2,2-difenil-1-picrilhidrazil) and FRAP (ferric reducing antioxidant power), and all of the fungi had positive results. The antimicrobial action of crude extracts had a good range of activities. All extracts had inhibitory activities against the yeasts of Candida albicans and C. parapsilosis, with 125 to 500 µg/mL MIC. Eight extracts had antimicrobial activities against Bacillus subtilis (MIC from 62.4 to 125 µg/mL), 5 against Pseudomonas aeruginosa (MIC from 125 to 500 µg/mL), 2 against Salmonella enterica (MIC from 125 to 62.5 µg/mL), and 2 against Enterococcus faecalis (MIC from 500 to 125 µg/mL). The presence of secondary metabolites, including coumarins, was observed during chemical evaluation by thin layer chromatography. Total phenol content was estimated, and a strong positive correlation to antioxidant activity was observed, according to its Pearson coefficient. This is the first report of the bioactive potential of endophytic fungi isolated from the Costaceae family in Brazilian ecosystems.

Prevalência de sepse e fatores de risco em neonatos de unidade de terapia intensiva de referência em Palmas,Tocantins, Brasil / Prevalence of sepsis and risk factors in newborns from intensive care unit of reference in Palmas, Tocantins, Brasil

As infecções microbianas neonatais, especialmente as sepses, correspondem a uma das causas mais importantes de morbimortalidade em unidades de terapia intensiva e sua prevalência é maior naqueles pacientes expostos a procedimentos invasivos. Objetivo: identificar a prevalência de sepse neonatal hospitalar, bem como os principais fatores de risco associados a esta infecção e os microrganismos envolvidos. Métodos: foi realizado um estudo retrospectivo revisando as fichas de investigação epidemiológica do Serviço de Controle de Infecção Hospitalar com dados dos recém-nascidos internados em unidade de terapia intensiva de referência estadual do Tocantins, no período de julho de 2007 a junho de 2009. Observou-se que 25% dos pacientes analisados desenvolveram sepse neste período, dos quais 60% foram do gênero masculino, 67% prematuros e 65% com peso ao nascer menor que 2500 gramas. Entre os fatores de risco de importância clínica, observou-se a prematuridade, baixo peso ao nascimento, ventilação mecânica, cateterismo venoso central e utilização de nutrição parenteral. A prematuridade e o peso ao nascimento foram fatores de menor risco relativo quando comparados aos demais fatores analisados. Resultados: foram identificados 111 pacientes com sepse, com taxa de mortalidade de 26%. Os principais microrganismos associados à sepse neonatal foram Klebsiella pneumoniae (28%), os Staphylococcus coagulase negativa - CoSN (20%) e Candida albicans (14%). Conclusão: os achados neste estudo reforçam a importância de ações de prevenção de infecções neonatais, bem como o manejo asséptico durante a instalação e a manutenção de procedimentos invasivos necessários aos cuidados intensivos neonatais

Neonatal bacterial infections, especially sepsis, are one of the most important causes of morbidity and mortality in intensive care units and they are more prevalent among patients who have undergone invasive procedures. Objective: to identify the prevalence of hospital neonatal sepsis as well as the associated risks factors for this infection and the involved microorganisms. Method: a retrospective study was done going over the epidemiological investigation forms of the Service of Hospital Infection Control with data from newborns admitted in a State reference intensive care unit in Tocantins, from July 2007 to June 2009. We noticed that 25% of the analyzed patients developed sepsis and 60% were males, 67% premature and 65% with a weight at birth below 2500 grams. Among the risks factors of clinical importance we noticed, the prematurity, low birth weight, mechanical ventilation, central venous catheterization and use of parenteral nutrition. The prematurity and birth weight were lower relative risks factors in comparison with the other analyzed factors. Results: 111 patients with sepsis were found, with a mortality rate of 26%. The main microorganisms associated with neonatal sepsis were Klebsiella pneumoniae (28%), Staphylococcus coagulase negative - CoSN (20%) and Candida albicans (14%). Conclusion: the findings on this study emphasize the importance of actions for prevention of neonatal infections as well as aseptic handling during set up and maintenance of invasive procedures needed in neonatal intensive care

The influence of carbohydrates in the interaction of Paracoccidioides brasiliensis with CCL-6 cells in vitro / A influência de carboidratos na interação entre Paracoccidioides brasiliensis e células CCL-6 (in vitro)

INTRODUCTION: Little is known about the early events in the interaction between Paracoccidioides brasiliensis and its host. To understand the effect of carbohydrates in the interaction between the fungus and epithelial cell in culture, we analyzed the influence of different carbohydrate solutions on the adhesion of P. brasiliensis yeast cells to CCL-6 cells in culture. METHODS: Fungal cells were cultivated with the epithelial cell line, and different concentrations of D-fucose, N-acetyl-glucosamine, D-mannose, D-glucosamine, D-galactosamine, sorbitol and fructose were added at the beginning of the experiment. Six hours after the treatment, the cells were fixed and observed by light microscopy. The number of P. brasiliensis cells that were adhered to the CCL-6 monolayer was estimated. RESULTS: The number of adhesion events was diminished following treatments with D-fucose, N-acetyl-glucosamine, D-mannose, D-glucosamine and D-galactosamine as compared to the untreated controls. Sorbitol and fructose-treated cells had the same adhesion behavior as the observed in the control. P. brasiliensis propagules were treated with fluorescent lectins. The FITC-labeled lectins WGA and Con-A bound to P. brasiliensis yeast cells, while SBA and PNA did not. CONCLUSIONS: The perceptual of adhesion between P. brasiliensis and CCL-6 cells decreased with the use of D-mannose, N-acetyl-glucosamine and D-glucosamine. The assay using FITC-labeled lectins suggests the presence of N-acetyl-glucosamine, α-mannose and α-glucose on the P. brasiliensis cell surface. An enhanced knowledge of the mediators of adhesion on P. brasiliensis could be useful in the future for the development of more efficient and less harmful methods for disease treatment and control.

INTRODUÇÃO: Pouco se conhece a respeito dos eventos iniciais que mediam as interações entre Paracoccidioides brasiliensis e seus hospedeiros. Com a intenção de compreender a importância de carboidratos junto a estas interações, foram analisados os efeitos de soluções de carboidratos sobre a adesão de células leveduriformes de P. brasiliensis sobre culturas de células CCL-6. MÉTODOS: As células fúngicas foram cultivadas com as células epiteliais e diferentes concentrações de D-fucose, N-acetyl-glucosamina, D-manose, D-glicosamina, D-galactosamina, sorbitol e frutose foram adicionadas ao cultivo no início da interação. Após 6h de tratamento, as células foram fixadas e observadas em microscópio óptico. RESULTADOS: Os tratamentos utilizando D-fucose, N-acetil-glicosamina, D-manose, D-glicosamina e D-galactosamina reduziram os números de adesões quando comparados com o controle. Os tratamentos realizados com o uso de sorbitol e frutose apresentaram os mesmos resultados observados no controle. Para detectar a presença de carboidratos na superfície do fungo, propágulos de P. brasiliensis foram tratados com lectinas fluorescentes. WGA-FITC e Con-A-FITC se ligaram às células de P. brasiliensis ao contrário de SBA e PNA. CONCLUSÕES: O percentual de adesão entre P. brasiliensis e células CCL-6 foi reduzido com o uso de D-manose, N-acetil-glicosamina e D-glicosamina. O uso de lectinas marcadas sugeriu a presença de N-acetil-glicosamina, α-manose e α-glicose na superfície de P. brasiliensis. Estes resultados contribuem para o aumento do conhecimento relacionado aos mediadores de adesão de P. brasiliensis, e poderão ser utilizados no futuro para o desenvolvimento de medidas mais eficientes para o controle e tratamento deste patógeno.

Endophytic fungi from plums (Prunus domestica) and their antifungal activity against Monilinia fructicola.

Enophytic fungi were isolated from plum (Prunus domestica) leaves, identified with ITS1 and ITS4 primers, and their antagonistic activity was tested against Monilinia fructicola, which causes brown rot, blossom blight, and twig blight of stone fruits, and Colletotrichum gloeosporioides, which causes anthracnose on a variety of fruit crops. The production of antifungal compounds was determined in agar-diffusion and volatile inverted-plate tests. A total of 163 fungi were recovered from 30 plum trees, representing 22 cultivars. Twenty-nine morphotypes were detected, but only 14 species were identified genetically. The most frequently isolated species was Phaeosphaeria nodorum, constituting 86.5% of the total isolates. Four isolates produced inhibitory volatiles to M. fructicola; however, no isolate produced volatiles inhibitory to C. gloeosporioides. The volatiles produced by these fungi were identified as ethyl acetate, 3-methyl-1-butanol, acetic acid, 2-propyn-1-ol, and 2-propenenitrile. The fungal volatiles inhibited growth and reduced width of the hyphae, and caused disintegration of the hyphal content. This is the first study describing fungal endophytes in stone fruits. The P. nodorum strains producing inhibitory volatiles could play a significant role in reduction of M. fructicola expansion in plum tissues. Potential of these strains for biological control of this pathogen on stone fruits warrants further investigation.

The influence of carbohydrates in the interaction of Paracoccidioides brasiliensis with CCL-6 cells in vitro.

INTRODUCTION: Little is known about the early events in the interaction between Paracoccidioides brasiliensis and its host. To understand the effect of carbohydrates in the interaction between the fungus and epithelial cell in culture, we analyzed the influence of different carbohydrate solutions on the adhesion of P. brasiliensis yeast cells to CCL-6 cells in culture. METHODS: Fungal cells were cultivated with the epithelial cell line, and different concentrations of D-fucose, N-acetyl-glucosamine, D-mannose, D-glucosamine, D-galactosamine, sorbitol and fructose were added at the beginning of the experiment. Six hours after the treatment, the cells were fixed and observed by light microscopy. The number of P. brasiliensis cells that were adhered to the CCL-6 monolayer was estimated. RESULTS: The number of adhesion events was diminished following treatments with D-fucose, N-acetyl-glucosamine, D-mannose, D-glucosamine and D-galactosamine as compared to the untreated controls. Sorbitol and fructose-treated cells had the same adhesion behavior as the observed in the control. P. brasiliensis propagules were treated with fluorescent lectins. The FITC-labeled lectins WGA and Con-A bound to P. brasiliensis yeast cells, while SBA and PNA did not. CONCLUSIONS: The perceptual of adhesion between P. brasiliensis and CCL-6 cells decreased with the use of D-mannose, N-acetyl-glucosamine and D-glucosamine. The assay using FITC-labeled lectins suggests the presence of N-acetyl-glucosamine, α-mannose and α-glucose on the P. brasiliensis cell surface. An enhanced knowledge of the mediators of adhesion on P. brasiliensis could be useful in the future for the development of more efficient and less harmful methods for disease treatment and control.

Enterobacteriaceae in mouth and cloaca of podocnemis expansa and P. Unifilis (testudines: chelonia) populations of national park of araguaia plains, Brazil

Shigella flexnerii and Escherichia coli were the most frequent Gram-negative bacteria found in the mouth cavity and cloacae of the turtles Podocnemis expansa and P. unifilis on beaches in the National Park of Araguaia, Brazil. Reptiles are known as Salmonella carriers, despite rarely isolated in these turtles.

Enterobacteriaceae in mouth and cloaca of podocnemis expansa and P. Unifilis (testudines: chelonia) populations of national park of araguaia plains, Brazil.

Shigella flexnerii and Escherichia coli were the most frequent Gram-negative bacteria found in the mouth cavity and cloacae of the turtles Podocnemis expansa and P. unifilis on beaches in the National Park of Araguaia, Brazil. Reptiles are known as Salmonella carriers, despite rarely isolated in these turtles.

Yeasts Occurring in Surface and Mouth Cavity of Two Chelonian Species, Podocnemis expansa Schweigger and P. unifilis Troschel (Reptilia: Chelonia: Pelomedusidae), in the Javaés River Border of Araguaia National Park in Brazil.

Thirty-eight specimens of free-ranging Podocnemis expansa (Amazon turtle) and 22 of P. unifilis (Tracajá) were screened for yeast isolation from surface (plastron, skin, and nails), eye, and mouth cavity. A hundred and eighteen yeast isolates belonging to 39 species were obtained. Debaryomyces hansenii, Candida galli, C. sake, and Rhodotorula mucilaginosa were the most frequent species isolated from these chelonians. Species diversity measured by Shannon's index was shown to be low and a degree of dominance could be detected as species known as potential pathogens were commonly isolated. The effective number of species in plastron of P. expansa was higher than in mouth samples, but not in P. unifilis probably due to dietary factors. P. expansa animals were captured on the beaches, and the superficial yeast populations may include terrestrial species. P. unifilis animals were captured in the water and the yeasts from superficial sites may represent species from river water.

Enterobacteriaceae associated with eggs of Podocnemis expansa and Podocnemis unifilis (Testudines:Chelonia) in nonpolluted sites of national park of Araguaia Plains, Brazil.

Fertile eggs of Podocnemis expansa and Podocnemis unifilis were investigated for the presence of enterobacteria, as these two endangered species have the potential for conservation measures that include egg transfer. Knowledge of normal microflora associated with turtles and turtle eggs would help effectively manage the transfer of these eggs among institutions. Thirty eggs of each species were collected, aseptically transferred, cracked inside plastic bags containing tetrathionate broth, and spread on selective media plates. Ten samples of sand and water were also examined for the presence of fecal coliforms using Colilert Quanti-Tray kits. Enterobacteriaceae were identified using an API 20E biochemical test kit. A majority of the bacteria isolated were potential pathogens. All egg samples were positive for Enterobacteriaceae, among which 15 eggs of P. expansa were positive for Shigella flexneri. Other isolates included Chromobacterium violaceum, Escherichia coli, and Aeromonas salmonicida subsp. salmonicida. Eggs of P. unifilis were most frequently positive for Shigella flexneri and C. violaceum, and less frequently positive for A. salmonicida subsp. salmonicida and Salmonella choleraesuis subsp. arizonae. The source of the bacteria in these eggs is unknown, but the nesting beaches were considered a source as water samples had low total coliform counts and E. coli was isolated from only one sand sample. This study demonstrated that Enterobacteriaceae are part of the indigenous microflora of chelonians.

Influência de Saccharomycopsis schoenii e Saccharomycopsis crataegensis na produção de aflatoxinas B1 e G1 por Aspergillus parasiticus em amendoim (Arachis hypogaea L. ) / Saccharomycopsis schoenii and Saccharomycopsis crataegensis effect on B1 and G1 aflatoxins production by Aspergillus parasiticus in peanut (Arachis hypogaea L. )

As aflatoxinas constituem o grupo de metabólitos secundários produzidos principalmente pelos Aspergillus flavus, A. parasiticus e A. nomius. Foi investigado o efeito isolado das leveduras Saccharomycopsis schoenii e S. crataegensis na produção de aflatoxinas B1 e G1 em amendoim, cultivar IAC Caiapó. As amostras de amendoim in natura e previamente autoclavadas foram inoculadas com A. parasiticus (1,6 x106 esporos.mL-1) e cultura das leveduras (1,6 x 108 células.mL-1), seguido de incubação a 25ºC durante sete dias. Foram realizados dois experimentos: no primeiro o fungo filamentoso e as leveduras foram inoculados simultaneamente. No segundo, a levedura foi inoculada 3 h antes da adição de fungo filamentoso. A quantificação das aflatoxinas foi executada por cromatografia em camada delgada. A produção das aflatoxinas B1 e G1 foi reduzida na presença das leveduras. A porcentagem de redução da concentração das aflatoxinas foi maior quando a suspensão do fungo foi adicionada 3 h após a inoculação da suspensão de leveduras. O decréscimo da concentração de aflatoxina B1 atingiu 89,3% e 82,6%, respectivamente na presença de S.schoenii e de S crataegensis. Os níveis de aflatoxina G1 foram reduzidos em 91,2% na presença de S.schoenii e em 93,2% quando S.crataegensis foi inoculada.