Long-term stability of hydrogen and organic acids production in an anaerobic fluidized-bed reactor using heat treated anaerobic sludge inoculum

This study evaluates the stability of hydrogen and organic acids production in an anaerobic fluidized-bed reactor (AFBR) that contains expanded clay (2.8-3.35 mm in diameter) as a support medium and is operated on a long-term basis. The reactor was inoculated with thermally pre-treated anaerobic sludge and operated with decreasing hydraulic retention time (HRT), from 8 h to 1 h, at a controlled temperature of 30 degrees C and a pH of about 3.8. Glucose (2000 mg L(-1)) was used as the substrate, generating conversion rates of 92-98%. Decreasing the HRT from 8 h to 1 h led to an increase in average hydrogen-production rates, with a maximum value of 1.28 L h(-1) L(-1) for an HRT of 1 h. In general, hydrogen yield production increased as HRT decreased, reaching 2.29 mol of H(2)/mol glucose at an HRT of 2 h and yielding a maximum hydrogen content of 37% in the biogas. No methane was detected in the biogas throughout the period of operation. The main soluble metabolites (SMP) were acetic acid (46.94-53.84% of SMP) and butyric acid (34.51-42.16% of SMP), with less than 15.49% ethanol. The steady performance of the AFBR may be attributed to adequate thermal treatment of the inoculum, the selection of a suitable support medium for microbial adhesion, and the choice of satisfactory environmental conditions imposed on the system. The results show that stable hydrogen production and organic acids production were maintained in the AFBR over a period of 178 days. (C) 2009 International Association for Hydrogen Energy. Published by Elsevier Ltd. All rights reserved.

Influence of a probiotic soy product on fecal microbiota and its association with cardiovascular risk factors in an animal model

Background: Previous work showed that daily ingestion of an aqueous soy extract fermented with Enterococcus faecium CRL 183 and Lactobacillus helveticus 416, supplemented or not with isoflavones, reduced the total cholesterol and non-HDL-cholesterol levels, increased the high-density lipoprotein (HDL) concentration and inhibited the raising of autoantibody against oxidized low-density lipoprotein (ox-LDL Ab) and the development of atherosclerotic lesions. Objective: The aim of this study was to characterize the fecal microbiota in order to investigate the possible correlation between fecal microbiota, serum lipid parameters and atherosclerotic lesion development in rabbits with induced hypercholesterolemia, that ingested the aqueous soy extract fermented with Enterococcus faecium CRL 183 and Lactobacillus helveticus 416. Methods: The rabbits were randomly allocated to five experimental groups (n = 6): control (C), hypercholesterolemic (H), hypercholesterolemic plus unfermented soy product (HUF), hypercholesterolemic plus fermented soy product (HF) and hypercholesterolemic plus isoflavone-supplemented fermented soy product (HIF). Lipid parameters and microbiota composition were analyzed on days 0 and 60 of the treatment and the atherosclerotic lesions were quantified at the end of the experiment. The fecal microbiota was characterized by enumerating the Lactobacillus spp., Bifidobacterium spp., Enterococcus spp., Enterobacteria and Clostridium spp. populations. Results: After 60 days of the experiment, intake of the probiotic soy product was correlated with significant increases (P < 0.05) on Lactobacillus spp., Bifidobacterium spp. and Enterococcus spp. and a decrease in the Enterobacteria population. A strong correlation was observed between microbiota composition and lipid profile. Populations of Enterococcus spp., Lactobacillus spp. and Bifidobacterium spp. were negatively correlated with total cholesterol, non-HDL-cholesterol, autoantibodies against oxidized LDL (ox-LDL Ab) and lesion size. HDL-C levels were positively correlated with Lactobacillus spp., Bifidobacterium spp., and Enterococcus spp. populations. Conclusion: In conclusion, daily ingestion of the probiotic soy product, supplemented or not with isoflavones, may contribute to a beneficial balance of the fecal microbiota and this modulation is associated with an improved cholesterol profile and inhibition of atherosclerotic lesion development.

Liquid-liquid extraction of proteases from fermented broth by PEG/citrate aqueous two-phase system

This work deals with the use of an aqueous two-phase system (ATPS) of PEG/citrate to remove proteases from a Clostridium perfringens fermentation broth. To plan the experimental tests and evaluate the corresponding results, three successive experimental designs were employed, for which the PEG molar mass (M-PEG) and concentration (C-PEG), the citrate concentration (C-C) and the pH were selected as independent variables, while the purification factor (PF), the partition coefficient (K), the activity yield (Y) and the selectivity (S) were selected as responses. PF of proteases in the top phase was shown to increase with increasing MPEG and decreasing Cc, whereas a completely opposite trend was observed for K. On the other hand, Y was favored by simultaneous decreases in both these variables, while S decreased with increasing Cc. Therefore, selecting a simultaneous increase in PF and Y as the most desirable result, the best performance of the system was obtained using M-PEG = 10-000 g/mol C-PEG = 22% (w/w) and C-c = 8.0% (w/w) at pH 8.5. Under these conditions, the activity yield was very high (131 %) but the purification factor (4.2) and the selectivity (4.3) were lower than those ensured by more selective purification methods. According to these results, the ATPS seems to be an interesting alternative primary concentration/decontamination step for vaccine preparation from C. perfringens fermented broth. (C) 2007 Elsevier B.V. All rights reserved.

Removal of a cysteine ligand from rubredoxin: assembly of Fe2S2 and Fe(S-Cys)(3)(OH) centres

The electron transfer protein rubredoxin from Clostridium pasteurianum contains an Fe(S-Cys)(4) active site. Mutant proteins C9G, C9A, C42G and C42A, in which cysteine ligands are replaced by non-ligating Gly or Ala residues, have been expressed in Escherichia coli. The C42A protein expresses with a (Fe2S2)-S-III cluster in place. In contrast, the other proteins are isolated in colourless forms, although a (Fe2S2)-S-III cluster may be assembled in the C42G protein via incubation with Fe-III and sulfide. The four mutant proteins were isolated as stable mononuclear Hg-II forms which were converted to unstable mononuclear Fe-III preparations that contain both holo and apo protein. The Fe-III systems were characterized by metal analysis and mass spectrometry and by electronic, electron paramagnetic resonance, X-ray absorption and resonance Raman spectroscopies. The dominant Fe-III form in the C9A preparation is a Fe(S-Cys)(3)(OH) centre, similar to that observed previously in the C6S mutant protein. Related centres are present in the proteins NifU and IscU responsible for assembly and repair of iron-sulfur clusters in both prokaryotic and eukaryotic cells. In addition to Fe(S-Cys)(3)(OH) centres, the C9G, C42G and C42A preparations contain a second four-coordinate Fe-III form in which a ligand appears to be supplied by the protein chain. Electronic supplementary material to this paper can be obtained by using the Springer Link server located at http://dx.doi.org/10.1007/s00775-0020355-1.

Autologous non-myeloablative haemopoietic stem cell transplantation in relapsing-remitting multiple sclerosis: a phase I/II study

Background Autologous non-myeloablative haemopoietic stem cell transplantation is a method to deliver intense immune suppression. We evaluated the safety and clinical outcome of autologous non-myeloablative haemopoietic stem cell transplantation in patients with retapsing-remitting multiple sclerosis (MS) who had not responded to treatment with interferon beta. Methods Eligible patients had relapsing-remitting MS, attended Northwestern Memorial Hospital, and despite treatment with interferon beta had had two corticosteroid-treated relapses within the previous 12 months, or one relapse and gadolinium-enhancing lesions seen on MRI and separate from the relapse. Peripheral blood haemopoietic stem cells were mobilised with 2 g per m(2) cyclophosphamide and 10 mu g per kg per day filgrastim. The conditioning regimen for the haemopoietic stem cells was 200 mg per kg cyclophosphamide and either 20 mg alemtuzumab or 6 mg per kg rabbit antithymocyte globulin. Primary outcomes were progression-free survival and reversal of neurological disability at 3 years post-transplantation. We also sought to investigate the safety and tolerability of autologous non-myeloablative haemopoietic stem cell transplantation. Findings Between January 2003, and February, 2005, 21 patients were treated. Engraftment of white blood cells and platelets was on median day 9 (range day 8-11) and patients were discharged from hospital on mean day 11 (range day 8-13). One patient had diarrhoea due to Clostridium difficile and two patients had dermatomal zoster. Two of the 17 patients receiving alemtuzumab developed late immune thrombocytopenic purpura that remitted with standard therapy. 17 of 21 patients (81%) improved by at least 1 point on the Kurtzke expanded disability status scale (EDSS), and five patients (24%) relapsed but achieved remission after further immunosuppression. After a mean of 37 months (range 24-48 months), all patients were free from progression (no deterioration in EDSS score), and 16 were free of relapses. Significant improvements were noted in neurological disability, as determined by EDSS score (p<0.0001), neurological rating scale score (p=0.0001), paced auditory serial addition test (p=0.014), 25-foot walk (p<0.0001), and quality of life, as measured with the short form-36 (SF-36) questionnaire (p<0.0001). Interpretation Non-myeloablative autologous haemopoietic stem cell transplantation in patients with relapsing-remitting MS reverses neurological deficits, but these results need to be confirmed in a randomised trial.

Effects of adding polyclonal antibody preparations on ruminal fermentation patterns and digestibility of cows fed different energy sources

Nine ruminally cannulated cows fed different energy sources were used to evaluate an avian-derived polyclonal antibody preparation (PAP-MV) against the specific ruminal bacteria Streptococcus bovis, Fusobacterium necrophorum, Clostridium aminophilum, Peptostreptococcus anaerobius, and Clostridium stick-landii and monensin (MON) on ruminal fermentation patterns and in vivo digestibility. The experimental design was three 3 x 3 Latin squares distinguished by the main energy source in the diet [dry-ground corn grain (CG), high-moisture corn silage (HMCS), or citrus pulp (CiPu)]. Inside each Latin square, animals received one of the feed additives per period [none (CON), MON, or PAP-MV]. Dry matter intake and ruminal fermentation variables such as pH, total short-chain fatty acids (tSCFA), which included acetate, propionate, and butyrate, as well as lactic acid and NH(3)-N concentration were analyzed in this trial. Total tract DM apparent digestibility and its fractions were estimated using chromic oxide as an external marker. Each experimental period lasted 21 d. Ruminal fluid sampling was carried out on the last day of the period at 0, 2, 4, 6, 8, 10, and 12 h after the morning meal. Ruminal pH was higher (P = 0.006) 4 h postfeeding in MON and PAP-MV groups when compared with CON. Acetate: propionate ratio was greater in PAP-MV compared with MON across sampling times. Polyclonal antibodies did not alter (P > 0.05) tSCFA, molar proportion of acetate and butyrate, or lactic acid and NH(3)-N concentration. Ruminal pH was higher (P = 0.01), 4 h postfeeding in CiPu diets compared with CG and HMCS. There was no interaction between feed additive and energy source (P > 0.05) for any of the digestibility coefficients analyzed. Starch digestibility was less (P = 0.008) in PAP-MV when compared with CON and MON. In relation to energy sources, NDF digestibility was greater (P = 0.007) in CG and CiPu vs. the HMCS diet. The digestibility of ADF was greater (P = 0.002) in CiPu diets followed by CG and HMCS. Feeding PAP-MV or monensin altered ruminal fermentation patterns and digestive function in cows; however, those changes were independent of the main energy source of the diet.

Flora bacteriana da cavidade oral, presas e veneno de Bothrops jararaca: possível fonte de infecção no local da picada

Materiais colhidos das presas, das bainhas das presas e do veneno de 15 Bothrops jararaca recém-capturadas, aparentemente saudáveis, foram submetidos a exame bacterioscópico e cultura aeróbia a anaeróbia. As bactérias mais freqüentemente isoladas foram os estreptococos do grupo D (1.2 serpentes), Enterobacter sp. (6), Providencia rettgeri (6), Providencia sp. (4), Escherichia coli (4), Morganella morganii (3) e Clostridium sp. (5). Como estas bactérias são semelhantes às encontradas nos abscessos de pacientes picados por serpentes do gênero Bothrops, é válido considerar a possibilidade de que bactérias da boca da serpente sejam inoculadas no momento da picada e, encontrando condições favoráveis de multiplicação, causem infecção.

Evolução do teor de nitritos e de nitratos e da concentração de pigmentos no fiambre e na mortadela ao longo do seu processo produtivo e do seu prazo de vida útil

Dissertação apresentada na Faculdade de Ciências e Tecnologia da Universidade Nova de Lisboa para a obtenção do grau de Mestre em Tecnologia e Segurança Alimentar

Relembrar o Botulismo

A propósito do seu caso pessoal, a autora tece algumas considerações acerca da intoxicação alimentar por toxina botulínica, chamando a atenção para cinco pontos principais: 1) O botulismo não é uma infecção tão rara como se pensa; 2) Está longe de ser uma doença com a mortalidade que geralmente se lhe atribui; 3) O veículo desta intoxicação não se limita às tradicionais conservas caseiras ou industriais; 4) É uma doença pouco diagnosticada, não apenas porque o seu diagnóstico é difícil mas sobretudo porque não se pensa nela; 5) Os doentes com botulismo são frequentemente rotulados com outros diagnósticos.

Botulism in Brazil, 2000-2008: epidemiology, clinical findings and laboratorial diagnosis

Botulism is a rare and potentially lethal illness caused by Clostridium botulinum neurotoxin. We describe the findings of a laboratorial investigation of 117 suspected cases of botulism reported to the surveillance system in Brazil from January 2000 to October 2008. Data on the number and type of samples analyzed, type of toxins identified, reporting of the number of botulism cases and transmission sources are discussed. A total of 193 clinical samples and 81 food samples were analyzed for detection and identification of the botulism neurotoxin. Among the clinical samples, 22 (11.4%) presented the toxin (nine type A, five type AB and eight with an unidentified type); in food samples, eight (9.9%) were positive for the toxin (five type A, one type AB and two with an unidentified type). Of the 38 cases of suspected botulism in Brazil, 27 were confirmed by a mouse bioassay. Laboratorial botulism diagnosis is an important procedure to elucidate cases, especially food-borne botulism, to confirm clinical diagnosis and to identify toxins in food, helping sanitary control measures.

An NMR structural study of nickel-substituted rubredoxin

J Biol Inorg Chem (2010) 15:409–420 DOI 10.1007/s00775-009-0613-6

Infecções Nosocomiais. Alguns Aspectos

A infecção nosocomial não é um problema recente, no entanto assume hoje em dia aspectos importantes, não só pela morbilidade e mortalidade associadas, mas também pelo seu dispêndio em recursos hospitalares. A maior susceptibilidade dos doentes em risco e o aparecimento de agentes cada vez mais resistentes aos antibióticos disponíveis são factores determinantes para a sua gravidade. O Staphylococcus aureus resistente à meticilina e o Clostridium difficile são dois dos agentes mais importantes pela sua frequência, gravidade das infecções que provocam e persistência. Actualmente são preocupação constante em hospitais por todo o mundo. O conhecimento da sua epidemiologia, particularidades clínico-patológicas e tratamento é fundamental para a vigilância e controle deste tipo de infecções por parte dos comités de infecção hospitalar. Por estas razões e por experiência recente com este tipo de infecções em dois hospitais dos E.U.A., me pareceu oportuna a sua abordagem.

Molecular determinants of ligand specificity in carbohydrate-binding modules: an NMR and X-ray crystallography integrated study

Dissertação para obtenção do Grau de Doutor em Bioquímica – Ramo Bioquímica Estrutural

Botulismo Infantil em Portugal – Um Lactente com Hipotonia

O Botulismo Infantil (BI) constitui uma síndrome neuroparalítica rara, potencialmente fatal, causada pela neurotoxina do Clostridium botulinum. Descreve-se o primeiro caso reportado desde o início da notificação obrigatória em Portugal (1999). Lactente de dois meses, internado por prostração, dificuldade alimentar e obstipação. Constatou-se envolvimento inicial dos pares cranianos associado a fraqueza muscular progressiva, descendente e simétrica. Constituíam factores de risco o consumo de mel caseiro e o banho com ervas de camomila. A confirmação diagnóstica foi efectuada pela pesquisa de esporos de Clostridium botulinum nas fezes e pela prova de inoculação em ratinhos. O tratamento implicou suporte respiratório e nutricional, e imunoglobulina humana anti-toxina, com evolução favorável. A clínica e o contexto epidemiológico são importantes para o diagnóstico, permitindo a instituição precoce do tratamento.

Surto epidêmico de gastrenterite, com identificação do provável alimento veiculador da infecção

Descreve-se um surto epidêmico de gastrenterite entre pessoas adultas, com um total de 52 casos, todos iniciando-se até 24 horas após terem ingerido um alimento provavelmente contaminado. Clinicamente, a doença caracterizou-se por dor abdominal em cólicas, urgência para evacuar, e missão de fezes diarreicas, febre e adinamia, que persistiram por um a tres dias. Todos os casos foram benignos, sem uma só complicação e nenhum tem necessidade de tratamento hospitalar. Foram feitos exames bacteriológicos das fezes de alguns casos, mas não se isolou qualquer germe; esses casos, porém, já estavam em convalescença ou em uso de antibióticos. Todos os casos tinham em comum o fato de terem almoçado ou feito lanche em um mesmo restaurante. Comparou-se, então, 109 pessoas que haviam almoçado, e 70 que haviam feito lanche, com 107 pessoas, do mesmo ambiente dos comensais, que não almoçaram nem lancharam. A associação com fato de almoçar ou lanchar no restaurante foi inequívoca. Ademais, em um número de pessoas bem maior do que os comensais, bebendo da mesma água, não surgiu um só caso de gastroenterite. As 109 pessoas que almoçaram responderam quais os alimentos que haviam ingerido, o que permitiu identificar a carne assada, servida no almoço, como sendo o veiculo provável da infecção. Infelizmente, não se conseguiu restos dessa carne assada para que se pudesse tentar isolar germes. A mediana do período de incubação foi 10,4 horas e os períodos de incubação máximo e mínimo foram de 24 e 0,5 horas, respectivamente; a distribuição cronológica dos casos lembra muito o perfil dos surtos epidêmicos de toxi-infecção alimentar produzidos pelo Clostridium perfrigens.