Production of Aspergillus niger GH1 tannase using solid-state fermentation

The production of tannase by Aspergillus niger GH1 in solid-state fermentation (SSF) was evaluated. Several concentrations of tannic acid (12.5, 25, 50 and 100 g/L) were tested in a model culture system. Column reactors were packed with polyurethane foam impregnated with liquid medium and inoculated with fungal spores. Tannase production was kinetically monitored by 48 h. Tannic acid uptake was spectrophotometrically measured and extracellular and intracellular tannase activities were assayed by HPLC-chromatography. Tannase activity increased with increasing concentration of tannic acid. Maximum extracellular and intracellular tannase activities (11.35 and 6.95 U/mL respectively) were recorded with 100 g/L of tannic acid. The substrate uptake was 100% at concentrations of 12.5, 25 and 50 g/L, while 74.4% was consumed in the presence of 100 g/L of tannic acid after 48 h of culture. These results suggest that high concentrations of tannins can be removed by SSF and tannase production can be reached in high levels.The present work was performed as part of a cooperative agreement between the Universidad Autonoma de Coahuila (UAdeC, Mexico) and the Universidade do Minho (UM, Portugal) within a specific international exchange program (VALNATURA project, alfa network from European Union) undertaken at the Biological Engineering Department (UM, Portugal) M A Cruz-Hernandez thanks to Concejo nacional de Ciencia y Tecnologia (CONACYT) the fellowship to study the doctoral program at the Department of Biotechnology (UAdeC

A turn-on fluorescent solid-sensor for Hg(II) detection

A rhodamine organosilane derivative (Rh-UTES) has been obtained by one-pot synthesis. The chemical structure of Rh-UTES was confirmed by nuclear magnetic resonance (NMR) and infrared (FTIR) techniques. To obtain an inorganic-organic hybrid sensor, Rh-UTES was covalently immobilized on a porous silicon microcavity (PSiMc) via triethoxysilane groups. The attachment of the organic derivative into PSiMc was confirmed by FTIR, specular reflectance, and scanning electron microscopy (SEM). The optical performance of Rh-UTES receptor for Hg2+ detection was investigated by fluorescent spectroscopy and microscopy. Upon the addition of increasing amounts of Hg2+ ions, a remarkable enhancement in emission intensity was produced in both systems. In the solid phase, an increase of integrated fluorescent emission of 0.12- and 0.15-fold after Hg2+ receptor coordination was observed. The light harvesting capability of PSiMc devices allowed obtaining an enhanced fluorescent emission after Rh-UTES immobilization (277-fold). The fluorescence microscopy of hybrid PSiMc sensor provided an optical qualitative test for Hg2+ detection.A rhodamine organosilane derivative (Rh-UTES) has been obtained by one-pot synthesis. The chemical structure of Rh-UTES was confirmed by nuclear magnetic resonance (NMR) and infrared (FTIR) techniques. To obtain an inorganic-organic hybrid sensor, Rh-UTES was covalently immobilized on a porous silicon microcavity (PSiMc) via triethoxysilane groups. The attachment of the organic derivative into PSiMc was confirmed by FTIR, specular reflectance, and scanning electron microscopy (SEM). The optical performance of Rh-UTES receptor for Hg2+ detection was investigated by fluorescent spectroscopy and microscopy. Upon the addition of increasing amounts of Hg2+ ions, a remarkable enhancement in emission intensity was produced in both systems. In the solid phase, an increase of integrated fluorescent emission of 0.12- and 0.15-fold after Hg2+ receptor coordination was observed. The light harvesting capability of PSiMc devices allowed obtaining an enhanced fluorescent emission after Rh-UTES immobilization (277-fold). The fluorescence microscopy of hybrid PSiMc sensor provided an optical qualitative test for Hg2+ detection

Preliminary culture conditions for Aspergillus niger GH1 tannase production in submerged culture

Nowadays, tannase is produce by submerged culture (SmC) being the enzyme obtained in an intracellular form resulting, as a consequence, in high production costs. To demonstrate that fungal tannase excretion is strongly influenced by the culture conditions, tannase production by Aspergillus niger GH1 on SmC using different bioreactors was evaluated. Two sets of experiments were conducted using in a first step a 1.5 L bioreactor and in a second step 100 mL erlenmeyer flasks. Modifications to culture conditions previously reported allowed to obtain positive results for tannase production in SmC, being the selected conditions: initial pH 5, temperature 35°C, initial substrate concentration 25g/L and agitation 200 rpm.A cultura submersa (SmC) é o processo empregado para produzir comercialmente a enzima tanasa, porém neste sistema da cultura, a tanasa é expressada principalmente de maneira intracelular o que representa custos de produção elevados. Para demonstrar que a excreção da tanasa do fungo está fortemente influenciada pelas condições da cultura, a produção da enzima tanase por Aspergillus niger GH1 em SmC foi avaliada. Dois conjuntos de experiências foram conduzidos usando em uma etapa um fermentador de 1.5 L e em uma segunda etapa matrazes (erlenmeyers) de 100 mL. As modificações às condições da cultura previamente descritas permitiram a obtenção de resultados positivos para a produção do tannase neste modo de produção, tendo sido definidas as seguintes condições de cultura: pH inicial 5, concentração inicial de substrato 25g/L, temperatura 35°C e agitação 200 rpm

Prevalence of Intestinal Parasites in Endangered Ashy Red Colobus Monkeys (Piliocolobus tephrosceles) in Tanzania

Intestinal parasites constitute one of the most frequent causes of gastrointestinal diseases in primates, directly affecting their health. We sampled 3 populations of the Endangered ashy red colobus monkey (Piliocolobus tephrosceles) with different levels of anthropogenic disturbance in Tanzania. We collected faecal samples (n=157) soon after defecation and fixed them in situ in 70% ethanol. We then re-fixed half of each sample in MIF (merthiolate iodine formaline) for microscopic study and saved the rest for molecular analysis. We examined helminth eggs, larvae and protozoan cysts using a light microscope after faecal sedimentation. We analysed samples positive for Giardia using Polymerase Chain Reaction (PCR) to determine genotypes. The overall prevalence of protozoan and helminth infection was 94.3% (148/157), with 64.9% (96/148) being infected by 1 species, 25.7% (38) by 2 species, and 9.5% (14) by 3 or more species. We detected 8 species of intestinal parasites: Ancylostoma sp. (13.4%), Trichuris trichiura (3.8%), Strongyloides stercoralis rhabditoid larvae (2.5%), Entamoeba chatonni (82.8%), Iodamoeba butschlii (14%), Endolimax nana (4.5%), Blastocystis hominis (2.5%) and Giardia duodenalis (14%). These species were detected in different combinations in the 3 areas, while Giardia was detected in only 1 area. The molecular analysis of positive Giardia samples showed that all of them belonged to assemblage B, which could also infect humans. However, we could not identify an exclusively anthropogenic origin of the parasitic species found. Our study contributes to our knowledge of parasitic infections in ashy monkeys in Tanzania, allowing us to assess their health status and disease risk, which in turn will help us design more successful conservation strategies for this Endangered primate in Tanzani

Spitzer Imaging of the Nearby Rich Young Cluster, Cep OB3b

We map the full extent of a rich massive young cluster in the Cep OB3b association with the IRAC and MIPS instruments aboard the {\it Spitzer} Space Telescope and the ACIS instrument aboard the $\it{Chandra}$ X-Ray Observatory. At 700 pc, it is revealed to be the second nearest large ($>1000$ member), young ($< 5$ Myr) cluster known. In contrast to the nearest large cluster, the Orion Nebula Cluster, Cep OB3b is only lightly obscured and is mostly located in a large cavity carved out of the surrounding molecular cloud. Our infrared and X-ray datasets, as well as visible photometry from the literature, are used to take a census of the young stars in Cep OB3b. We find that the young stars within the cluster are concentrated in two sub-clusters; an eastern sub-cluster, near the Cep B molecular clump, and a western sub-cluster, near the Cep F molecular clump. Using our census of young stars, we examine the fraction of young stars with infrared excesses indicative of circumstellar disks. We create a map of the disk fraction throughout the cluster and find that it is spatially variable. Due to these spatial variations, the two sub-clusters exhibit substantially different average disk fractions from each other: $32$ and $50$. We discuss whether the discrepant disk fractions are due to the photodestruction of disks by the high mass members of the cluster or whether they result from differences in the ages of the sub-clusters. We conclude that the discrepant disk fractions are most likely due to differences in the ages.Comment: 48 Pages, 12 figures, 6 table

Gallic acid production with mouldy polyurethane particles obtained from solid state culture of Aspergillus niger GH1

Gallic acid production in a batch bioreactor was evaluated using as catalytic material the mouldy polyurethane solids (MPS) obtained from a solid-state fermentation (SSF) bioprocess carried out for tannase production by Aspergillus niger GH1 on polyurethane foam powder (PUF) with 5 % (v/w) of tannic acid as inducer. Fungal biomass, tannic acid consumption and tannase production were kinetically monitored. SSF was stopped when tannase activity reached its maximum level. Effects of washing with distilled water and drying on the tannase activity of MPS were determined. Better results were obtained with dried and washed MPS retaining 84 % of the tannase activity. Maximum tannase activity produced through SSF after 24 h of incubation was equivalent to 130 U/gS with a specific activity of 36 U/mg. The methylgallate was hydrolysed (45 %) in an easy, cheap and fast bioprocess (30 min). Kinetic parameters of tannase self-immobilized on polyurethane particles were calculated to be 5 mM and 04.1×102 mM/min for K M and V max, respectively. Results demonstrated that the MPS, with tannase activity, can be successfully used for the production of the antioxidant gallic acid from methyl-gallate substrate. Direct use of PMS to produce gallic acid can be advantageous as no previous extraction of enzyme is required, thus reducing production costs.Authors thank the National Council for Science and Technology (CONACYT-Mexico) for the financial support. The present work was performed as part of a cooperative agreement between DIA-Universidad Autonoma de Coahuila (Mexico) and IBB-Universidade do Minho (Portugal) within a specific training stay undertaken at the DEB-UM. Part of the research was funded by a project SEP-CONACYT-CB-2011

Human Milk Antioxidative Modifications in Mastitis: Further Beneficial Effects of Cranberry Supplementation

Mastitis is the inflammation of one or several mammal lobes which can be accompanied by a mammary gland infection, and is the leading cause of undesired early weaning in humans. However, little information exists regarding the changes that this disease may induce in the biochemical composition of human milk, especially in terms of oxidative status. Given that newborns are subject to a significant increase in total ROS burden in their transition to neonatal life and that their antioxidant defense system is not completely developed, the aim of this study was to evaluate antioxidant defense (glutathione peroxidase (GPx), reduced glutathione (GSH), total polyphenol content (TPP), and total antioxidant capacity (TAC)) in milk samples from mothers suffering from mastitis and controls. We also measured the oxidative damage to lipids (malondyaldehyde (MDA)) and proteins (carbonyl group content (CGC)) in these samples. Finally, we tested whether dietary supplementation with cranberries (a product rich in antioxidants) in these breastfeeding mothers during 21 days could improve the oxidative status of milk. GPx activity, TPP, and TAC were increased in milk samples from mastitis-affected women, providing a protective mechanism to the newborn drinking mastitis milk. MDA concentrations were diminished in the mastitis group, confirming this proposal. Some oxidative damage might occur in the mammary gland since the CGC was increased in mastitis milk. Cranberries supplementation seems to strengthen the antioxidant system, further improving the antioxidative state of milk

Chimpanzee vertebrate consumption: Savanna and forest chimpanzees compared

There is broad consensus among paleoanthropologists that meat-eating played a key role in the evolution of Homo, but the details of where, when, and why are hotly debated. It has been argued that increased faunivory was causally connected with hominin adaptation to open, savanna habitats. If savanna-dwelling chimpanzees eat meat more frequently than do forest chimpanzees, it would support the notion that open, dry, seasonal habitats promote hunting or scavenging by hominoids. Here we present observational and fecal analysis data on vertebrate consumption from several localities within the dry, open Ugalla region of Tanzania. Combining these with published fecal analyses, we summarize chimpanzee vertebrate consumption rates, showing quantitatively that savanna chimpanzee populations do not differ significantly from forest populations. Compared with forest populations, savanna chimpanzees consume smaller vertebrates that are less likely to be shared, and they do so more seasonally. Analyses of chimpanzee hunting that focus exclusively on capture of forest monkeys are thus difficult to apply to chimpanzee faunivory in open-country habitats and may be misleading when used to model early hominin behavior. These findings bear on discussions of why chimpanzees hunt and suggest that increases in hominin faunivory were related to differences between hominins and chimpanzees and/or differences between modern and Pliocene savanna woodland environments