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Doctoral Thesis
DOI
https://doi.org/10.11606/T.18.2009.tde-29042009-105734
Document
Author
Full name
Sandra Imaculada Maintinguer
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Carlos, 2009
Supervisor
Committee
Silva, Maria Bernadete Amâncio Varesche (President)
Arenas, Claudia Etchebehere
Foresti, Eugenio
Oliveira, Valéria Maia de
Sakamoto, Isabel Kimiko
Title in Portuguese
Obtenção e caracterização filogenética de consórcio bacteriano utilizado em reator anaeróbio em batelada aplicado à produção de hidrogênio
Keywords in Portuguese
Clostridium sp.
Fermentação
Hidrogênio
Sacarose
Xilose
Abstract in Portuguese
O pré-tratamento a quente do inoculo associado ao controle do pH tem sido aplicado para selecionar bactérias produtoras de hidrogênio, como por exemplo, Clostridium sp. Nesse sentido, o objetivo desse trabalho foi obter consórcio de bactérias anaeróbias produtoras de hidrogênio a partir de inóculo proveniente de lodo granulado de reator UASB, usado no tratamento de dejetos de suinocultura. Para tanto, foi realizado tratamento a quente do inóculo e, posterior, enriquecimento com diluições seriais visando obter consórcio anaeróbio. Foram utilizados 20% (v/v) desse consórcio como inóculo nos ensaios de determinação de gás hidrogênio em reatores com diferentes concentrações de sacarose e xilose. Os reatores anaeróbios (2 L) foram alimentados com sacarose e xilose, separadamente, em triplicata, a 37 graus Celsius, pH 5,5 e headspace com 'HE' (100%) para as seguintes condições: (1) 630,0 mg sacarose/L; (2) 1184,0 mg sacarose/L; (3) 1816,0 mg sacarose/L, (4) 4128,0 mg sacarose/L, (5) 630,0 mg xilose/L; (6) 1341,0 mg xilose/L; (7) 1848,0 mg xilose/L e (8) 3588,0 mg xilose/L. A produção de 'H IND.2' foi verificada em todos os reatores anaeróbios e não houve geração de metano. Os rendimentos de substrato a 'H IND.2' para as condições (1), (2), (3), (4), (5), (6), (7) e (8) foram, respectivamente, 15% (1,2 mol 'H IND.2'/mol sacarose), 20% (1,6 mol 'H IND.2'/mol sacarose), 15% (1,2 mol 'H IND.2'/mol sacarose), 4% (0,3 mol 'H IND.2'/mol sacarose); 3,0% (0,2 mol 'H IND.2'/mol xilose), 8,0% (0,5 mol 'H IND.2'/mol xilose), 10,0% (0,6 mol 'H IND.2'/mol xilose) e, 14% (0,8 mol 'H IND.2'/mol xilose). Em todos os reatores foram detectados os ácidos acético e butírico e etanol. A morfologia predominante foi de bacilos Gram positivos e formadores de endósporos. Análises de clonagem e sequenciamento do consórcio bacteriano revelaram semelhanças com Clostridium, Burkolderia, Klebsiella e bactérias não cultivadas. A produção biológica de gás hidrogênio foi devida a consórcios de bactérias que estiveram presentes nas condições estudadas, identificados, principalmente, por espécies de, Clostridium sp., Clostridium acidotolerans e Klebsiella pneumoniae, reconhecidas como geradoras de 'H IND.2' e ácidos voláteis.
Title in English
Collection and phylogenetic characterization of bacterial consortium used in batch anaerobic reactor applied to hydrogen production
Keywords in English
Clostridium sp.
Fermentation
Hydrogen
Sucrose
Xylose
Abstract in English
The heat treatment of the inoculum associated with the pH control has been applied to select hydrogen-producing bacteria such as Clostridium sp. Therefore, this study aimed at preserving and keeping the anaerobic bacteria cultures producing hydrogen that comes from the granular sludge in UASB reactor, used to treat waste from pig farming, after heat treatment. For this, heat treatment of the inoculum was done and later, enrichment with serial dilutions to get anaerobic consortium. The 20% (v/v) of the bacterial consortium was used in essays for determination of hydrogen gas in reactors with different concentrations of sucrose and xylose. The reactors (2 L), fed with sucrose and xylose, separately, in triplicate, to 37 Celsius degrees, pH 5.5 and headspace with 'HE' (100%) for the conditions: (1) 630.0 mg sucrose/L, (2) 1184.0 mg sucrose/L, (3) 1816.0 mg sucrose/L, (4) 4128.0 mg sucrose/L, (5) 630.0 mg xylose/L, (6) 1341.0 mg xylose /L, (7) 1848.0 mg xylose/L and (8) 3588.0 mg xylose/L. 'H IND.2' production was detected in all the anaerobic reactors and there was no methane generation. 'H IND.2' yield for the conditions (1), (2), (3), (4), (5), (6), (7) and (8) were, respectively, 15% (1, 2 mol 'H IND.2'/mol sucrose), 20% (1.6 mol 'H IND.2'/mol sucrose), 15% (1.2 mol 'H IND.2'/mol sucrose), 4% (0.3 mol 'H IND.2'/mol sucrose), 3.0% (0.2 mol 'H IND.2'/mol xylose), 8.0% (0.5 mol 'H IND.2'/mol xylose), 10.0% (0.6 mol 'H IND.2'/mol xylose), and 14% (0.8 mol 'H IND.2'/mol xylose). The metabolic intermediarie products were acetic, butyric acid and ethanol in all anaerobic reactors. The main morphology was Gram positive bacilli and endospores formers. The cloning and sequencing analysis of the bacterial consortium revealed the presence of Clostridium genus (57%), Burkolderia genus (30%), Klebsiella genus (8%) and uncultivated bacteria (5%). The hydrogen gas production was due to bacterial consortium that were present in the studied conditions, marked mainly by species Clostridium sp., Clostridium acidotolerans and Klebsiella pneumoniae, recognized as 'H IND.2' and volatile acids producing.
 
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SandraMaintinguer.pdf (1.33 Mbytes)
Publishing Date
2009-05-05
 
WARNING: The material described below relates to works resulting from this thesis or dissertation. The contents of these works are the author's responsibility.
  • MAINTINGUER, S. I., et al. Fermentative hydrogen production by microbial consortium. International Journal of Hydrogen Energy, 2008, vol. 33, p. 4309-4317.
  • Maintinguer, Sandra I., et al. Evaluation of the microbial diversity of denitrifying bacteria in bactch reactor. Brazilian Journal of Chemical Engineering , 2013, vol. 30, p. 457-465.
  • Maintinguer, Sandra I., et al. Fermentative hydrogen production with xylose by Clostridium and Klebsiella species in anaerobic batch reactors [doi:10.1016/j.ijhydene.2011.07.095]. International Journal of Hydrogen Energy [online], 2011, vol. 36, p. 13508-13517.
  • MAINTINGUER, S. I., et al. Avaliação da desnitrificação de amostra de sistema de lodos ativados sob diferentes relações carbono e nitrogênio. In XVII Simpósio Nacional de Bioprocessos - SINAFERM, Natal/RN, 2009. XVII SINAFERM., 2009.
  • MAINTINGUER, S. I., et al. Biological hydrogen production from environmental sample in tropical countries. In Simpósio de Microbiologia Aplicada, Rio Claro - SP, 2011. Simpósio de Microbiologia Aplicada., 2011. Abstract.
  • MAINTINGUER, S. I., et al. Diversity of anaerobic bacterial communities in natural environments. In 13 World Congress on Anaerobic Digestion - Recovering (bio) Resources for the World, Santiago de Compostela, 2013. Proceedings of 13th World Congress on Anaerobic Digestion., 2013.
  • MAINTINGUER, S. I., et al. Evaluation of the microbial diversity from environmental sample applied to hydrogen bioproduction. In Fourth International Symposium on Energy fromBiomass and Waste, Veneza, 2012. Fourth International Symposium on Energy fromBiomass and Waste., 2012.
  • MAINTINGUER, S. I., et al. Evaluation of the microbial diversity in anaerobic reactors fed with sucrose applied to hydrogen production. In II International Conference on Environmental, Industrial and Applied Microbiology, Seville - Espanha, 2007. BioMicroWorld - 2007., 2007. Available from: http://www.formatex.org/biomicroworld2007.
  • MAINTINGUER, S. I., et al. Hydrogen biologic production from environmental sample in tropical countries. In 6th Annual International Symposium on Environment, Athenas - Grécia, 2011. 6th Annual International Symposium on Environment., 2011.
  • MAINTINGUER, S. I., et al. Purificação de bactérias anaeróbias produtoras de hidrogênio. In Sinaferm 2007 - XVI Simpósio Nacional de Bioprocessos, Curitiba - Paraná, 2007. Sinaferm 2007 - XVI Simpósio Nacional de Bioprocessos., 2007. Dispon?vel em: http://sinaferm 2007 - xvi simp?sio nacional de bioprocessos.
  • MAINTINGUER, S. I., SAKAMOTO, I. K., and Varesche, M. B. Evaluation of microbial diversity of denitrifying bacteria in activated sludge system. In The Water Research Conference, Lisboa - Portugal, 2010. The Water Research Conference., 2010.
  • MAINTINGUER, S. I., SAKAMOTO, I. K., e VARESCHE, M. B. A. Avaliação da comunidade e arquéias metanogênicas em sedimento de reservatório. In X Latin American Workshop and Symposium on Anaerobic Digestion, Ouro preto - MG, 2011. X DAAL., 2011.
  • Maintinguer, Sandra Imaculada, Sakamoto, Isabel Kimiko, e Varesche, M. B. Avaliação da diversidade microbiana de reator anóxico em batelada. In XX Congresso Latinoamericano de Microbiología, Montevideo - Uruguay, 2010. XX Congresso Latinoamericano de Microbiología., 2010. Resumo.
  • PACHIEGA, R., et al. Consortium of bacteria producing hydrogen from low sugars concentration. In 13 World Congress on Anaerobic Digestion - Recovering (bio) Resources for the World, Santiago de Compostela, 2013. Proceedings of 13th World Congress on Anaerobic Digestion., 2013.
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