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Master's Dissertation
DOI
10.11606/D.9.2016.tde-12082016-091342
Document
Author
Full name
Elizandra Hertel Lenhardt
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2016
Supervisor
Committee
Ishii, Marina (President)
Penna, Thereza Christina Vessoni
Matsudo, Marcelo Chuei
Title in Portuguese
Avaliação da produção e viabilidade de esporos de Bacillus atrophaeus ATCC 9372 utilizando resíduos do processamento de suco de laranja
Keywords in Portuguese
Esporos de Bacillus atrophaeus
Indicador biológico
Microbiologia aplicada
Processamento de laranja
Resíduos agroindustriais
Resistência térmica
Valor D
Abstract in Portuguese
O Brasil é um dos maiores produtores mundiais de suco de laranja, da mesma forma que a produção é elevada, a geração de resíduos também é significativa. Sabe-se que estes resíduos, os quais incluem sementes, cascas e restos de polpa são ricos em nutrientes que poderiam ser utilizados como substrato por micro-organismos, seja para o crescimento ou para a obtenção de subprodutos. Esporos de Bacillus atrophaeus ATCC 9372 são utilizados como indicadores biológicos, IBs, em processos térmicos por formarem esporos termorresistentes. O objetivo deste trabalho foi avaliar o uso de resíduos do processamento de suco de laranja como um meio de cultura alternativo para obtenção de esporos de B. atrophaeus, para serem aplicados em processos industriais. Ao bagaço de laranja (de 1,0 g a 20,0 g), obtido por processamento em centrífuga de frutas, foram adicionados 100 mL de água, e incubados a 150 rpm / 37 ºC por até 6 dias. Evidenciada a viabilidade de crescimento celular (µmáx = 0,0238 h-1 e Px = 0,0787 g/L.h, para 5,0 g de bagaço) procedeu-se ao estudo de planejamento experimental fatorial 22 em formato estrela com 6 pontos centrais, considerando a concentração de bagaço e o volume de meio. Foram determinados os valores de pH, de biomassa, de esporos viáveis e a resistência térmica dos mesmos a 102 ºC. Observou-se que houve aumento nos valores de pH após o cultivo e que as maiores concentrações de esporos foram de 1,73 x 109 esporos /mL e 5,75 x 109 esporos /mL após 3 e 6 dias de cultivo e os tempos de redução decimal determinados variaram de D102C = 0,92 min a D102C = 2,71 min e de D102C = 1,34 min a D102C = 3,98 min após 3 e 6 dias de cultivo, respectivamente. Com base no planejamento proposto e a análise de regressão, o desenvolvimento de esporos em bagaço segue a relação: Esporos = {-1,15 + 0,0303* [bagaço (g)] - 0,00611* [volume (mL)] + 0,611* [tempo (dias)]}, p=0,000, R2 =0,452, sendo o tempo (p=0,000) o fator de maior influência na formação de esporos. Os meios preparados com bagaço de laranja apresentaram-se viáveis para a produção de esporos de B. atrophaeus termorresistentes, produto de interesse farmacêutico e industrial, agregando valor ao resíduo que seria descartado.
Title in English
Evaluation of production and viability of Bacillus atrophaeus ATCC 9372 spores using orange juice processing waste
Keywords in English
Agricultural residues
Applied microbiology
Bacillus atrophaeus spores
Biological indicator
D value
Processing of orange
Thermal resistance
Abstract in English
Brazil is one of the world´s largest producers of oranges juice, in the same way that the production is high the amount of generated waste is also significant. It is well known that these residues, which include seeds, peel and pulp, are rich in nutrients that could be used as substrate by microorganisms whether for growth or for obtaining by-products. Bacillus atrophaeus ATCC 9372 spores are used as biological indicators, BIs, in thermal processes due to their ability to form heat-resistant spores. This study aimed to evaluate the use of orange juice processing waste as an alternative culture media to obtain B. atrophaeus spores, to be applied in industrial processes. To orange's bagasse (from 1.0 g to 20.0 g), obtained by processing in a fruit's centrifuge, 100 mL of water was added, and sterilized at 121 ºC. An aliquot of 0.1g/L of Bacillus atrophaeus spores was inoculated to bagasses's media and incubated at 150 rpm / 37 ºC up to 6 days. As cells (µmáx = 0.0238 h-1 and Px = 0.0787 g/L.h, for 5.0 g of bagasse) were obtained, a factorial experimental design 22, with star-shaped model and 6 central points, was performed considering the bagasse concentration and the media volume used. Values of pH, biomass, viable spores and their thermal resistance at 102 ºC were determined. It was observed that pH increased after cultivation and major values of spore concentration achieved were 1.73 x 109 spores /mL and 5.75 x 109 spores /mL after 3 and 6 days, respectively. Decimal reduction times determined ranged from D102C = 0.92 min to D102C = 2.71 min and from D102C = 1.34 min to D102C = 3.98 min after 3 and 6 days of incubation, correspondingly. The regression analysis showed that the development of spores in bagasse can be defined by the equation: Spores = {-1.15 + 0.0303* [bagasse (g)] - 0.00611* [volume (mL)] + 0.611* [time (days)]}, p=0.000, R2 =0.452 and time has a positive influence in the spore formation. Results demonstrated media prepared with oranges' bagasse were capable to grow and to develop B. atrophaeus heat-resistant spores, being an alternative to add value to a waste that would be discarded, generating a product of great importance in the pharmaceutical field.
 
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Publishing Date
2016-09-02
 
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