Poultry slaughterhouse anaerobic ponds as a source of inoculum for biohydrogen production.

Several waste sources have been studied as substrate sources for the production of biogas rich in hydrogen and for the isolation of bacteria capable of fermenting several substrates for the same purpose. Nonetheless, to simplify the process and minimize production costs, it is important to seek alternatives both for the use of microbial consortia using crude waste and for the use of substrates also in their crude form, without the need for purification. The aim of this study was to use only waste as inoculum and substrate for the biological production of hydrogen. Thus, samples from anaerobic ponds of a poultry slaughterhouse were used as inoculum. Sucrose, pure glycerol (in initial tests) and crude glycerol (inserted in blends with pure glycerol) were used as substrates. H2 production experiments were conducted in batches, using a reactor kept in an anaerobic environment for 11 days, at 35°C, under orbital agitation at 150 rpm. To analyse the composition of the biogas and the presence of soluble metabolic products (SMPs), samples of the headspace gases generated and of the reaction medium were collected. The results using sucrose as substrate indicated that the inoculum under study has potential for bio-H2 production, as it produced CH4-free biogas containing 50-60% H2. The inoculum was also shown to be adaptable to the use of glycerine as a substrate, producing biogas with similar characteristics to those obtained from sucrose degradation; however, it required a longer acclimatization period, and thus more in-depth study is required.

Fucogalactan from the giant mushroom Macrocybe titans inhibits melanoma cells migration.

An aqueous extract containing polysaccharides was obtained from the giant mushroom Macrocybe titans, and it was purified by amylase treatment, freeze-thawing process and dialysis. The purified fraction (ESP) was analyzed by HPSEC and GC-MS which showed a homogenous polysaccharide with Mw 14.2 × 103 g/mol composed by galactose and fucose. NMR and methylation analysis of ESP confirmed the presence of a fucogalactan with a (1 → 6)-linked α-d-Galp main chain partially substituted at O-2 by non reducing end units of α-l-Fucp residues in the side chain. Its biological activity was evaluated against murine melanoma cells B16-F10. The fucogalactan did not alter the viability, proliferative capacity and morphology of cells. However, this polysaccharide was able to reduce the cell migration in vitro at 40% (100 µg/mL) and 33% (250 µg/mL). The results obtained showed that Macrocybe titans fucogalactan is a promising agent capable of altering melanoma cell migration without decrease the cell viability.

Candida albicans PROTEIN PROFILE CHANGES IN RESPONSE TO THE BUTANOLIC EXTRACT OF Sapindus saponariaL.

Candida albicans is an opportunistic human pathogen that is capable of causing superficial and systemic infections in immunocompromised patients. Extracts of Sapindus saponaria have been used as antimicrobial agents against various organisms. In the present study, we used a combination of two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) to identify the changes in protein abundance of C. albicans after exposure to the minimal inhibitory concentration (MIC) and sub-minimal inhibitory concentration (sub-MIC) of the butanolic extract (BUTE) of S. saponaria and also to fluconazole. A total of six different proteins with greater than 1.5 fold induction or repression relative to the untreated control cells were identified among the three treatments. In general, proteins/enzymes involved with the glycolysis (GPM1, ENO1, FBA1), amino acid metabolism (ILV5, PDC11) and protein synthesis (ASC1) pathways were detected. In conclusion, our findings reveal antifungal-induced changes in protein abundance of C. albicans. By using the previously identified components of the BUTE of S. saponaria(e.g., saponins and sesquiterpene oligoglycosides), it will be possible to compare the behavior of compounds with unknown mechanisms of action, and this knowledge will help to focus the subsequent biochemical work aimed at defining the effects of these compounds.

Diagnosis of American cutaneous leishmaniasis by enzyme immunoassay using membrane antigens of Leishmania (Viannia) braziliensis.

This study evaluated the reactivity of membrane antigens of Leishmania (Viannia) braziliensis for the diagnosis of ACL by enzyme immunoassay (EIA). Promastigotes of L. (V.) braziliensis were grown in medium 199 and lysed in a sonicator. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting showed that specific proteins of L. (V.) braziliensis (apparent molecular weights 36 kDa and 48-56 kDa) were recognized by sera from ACL patients. These proteins were eluted from the SDS-PAGE and tested in EIA-IgG with sera from ACL patients, healthy individuals, patients with toxoplasmosis, paracoccidioidomycosis, syphilis, tuberculosis, leprosy, and Chagas disease. The EIA-IgG with membrane antigens allowed us to distinguish patients with ACL from healthy individuals and patients with other diseases (P < 0.0001), and showed a sensitivity of 93.3% and specificity of 90.8%, not including Chagas disease patients. 2D-SDS-PAGE followed by Western blotting was performed to improve the characterization of the antigens, and showed a component with isoelectric points near the acid pH side and apparent molecular weights of 48-56 kDa. The results showed good sensitivity and specificity of EIA-IgG with membrane antigens, indicating their potential use for diagnosis of ACL, as well as seroepidemiological surveys and follow-up of clinically cured patients.

Biomass and exopolysaccharide production in submerged cultures of Pleurotus ostreatoroseus Sing. and Pleurotus ostreatus "florida" (Jack.: Fr.) Kummer.

The species of the genus Pleurotus produce large amounts of biomass and exopolysaccharide (EPS) in submerged cultures, which may be used for biotechnological purposes. In the present work two Brazilian autochthonous strains of edible Pleurotus (P. ostratoroseus Sing. and P. ostreatus "florida") were used. The fungi grown in liquid Potato Dextrose medium (PD) were used as inocula to cultivate those microorganisms in the POL culture medium. After a 9-day incubation, the optimal growth time for biomass production, P. ostreatus "florida" presented higher biomass production (22.8 g d.w./l) than P. ostreatoroseus (16.8 g d.w./l). After a 7-day incubation, the optimal growth time for EPS production, P. ostreatoroseus produced higher amounts of crude EPS (5.8 g d.w./l) than P. ostreatus "florida" (1.4 g d.w./l). Relative carbohydrate composition for P. ostreatoroseus and P. ostreatus "florida" EPS were: glucose (95.5-87.7), galactose (traces - 4.9), mannose (traces - 3.1), xylose (1.3-2.5), and arabinose (3.2-1.8). Lower ammonium sulfate concentration in the POL culture medium increased the exopolysaccharides production by P. ostreatoroseus.

Alternative method of inoculum and spawn production for the cultivation of the edible brazilian mushroom Pleurotus ostreatoroseus SING.

Efficiency of solid and liquid inocula and their use for spawn production were compared so that improved cultivation conditions for the edible mushroom Pleurotus ostreatoroseus could be tested. Solid and liquid inocula were prepared respectively with Potato Dextrose Agar (PDA) and Liquid Potato Dextrose (LPD). Wheat grains and cotton residues were used as substrates for spawn preparation. Inoculum types did not affect the development of P. ostreatoroseus, and LPD spawns were cheaper, more homogenous, less contaminated. Decomposition activity of mushroom growth, as a percentage of organic matter loss (OML), was higher in the wheat grain spawn and was not influenced by the inoculum type. Advantages in the use of cotton residue for spawn production were longer storage time, lower contamination and reduced costs. The cotton residue substrate may be also used for the production of mushroom fruiting bodies.