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Comparative analysis of biohydrogen producing bacterial consortia in three thermophilic anaerobic fluidised bed bioreactors

A research report submitted to the Faculty of Health Sciences, University of the
Witwatersrand, in partial fulfilment of the requirements for the degree of Master
of Science in Medicine (Pharmaceutical Affairs).
Johannesburg 2012 / Global warming has stimulated research into alternative energy carries and fuels. Hydrogen is
one of these alternative fuels that are recognized as a promising future energy source.
Historically, it is produced by water electrolysis and the gasification of coal. Hydrogen is a
natural though transient by-product of several microbial driven biochemical reactions,
including anaerobic digestion and fermentation. Microorganisms degrade complex molecules
to produce butyrate and alcohols with CO2 and hydrogen as the only by- products. Hydrogen
produced by microorganisms is known as biohydrogen.
This study aimed to identify biohydrogen- producing bacteria in three Anaerobic Fluidised
Bed Bioreactors (AFBRs), which are capable of producing hydrogen under anaerobic and
thermophilic conditions, by using PCR-DGGE analysis of the 16 S rDNA genes. Sewage
sludge from anaerobic digester and cow- dung were used as inoculum to isolate potential H2-
producing organisms. The operational conditions were manipulated to removing mesophilic
bacteria and non- spore forming bacteria by gradually increasing the temperature to
thermophilic conditions and the pH maintained at acid conditions to allow acidotolerant
bacteria to survive. The bioreactors were operated for a period of forty days for each research
cycle. Bioreactor One was operated at 55°C for eight days and then the temperature kept at a
constant 65°C for the remainder of the research cycle. Bioreactor Two was operated for 8
days at each temperature ranging from 45°C - 65°C, with increments of 5 °C. Bioreactor
Three was operated at each temperature for ten days ranging from 55- 70°C, also with
increments of 5°C. Samples for microbial community identification were taken at 55°C and
65°C. The bacterial morphologies and structural properties were evaluated by examining the
hydrogen- producing granules, isolated at 65°C, using scanning electron microscopy. Species
of the families Bacillus, Enterobacteria, Actinomyces, Clostridium and Veillonella were
identified. Clostridium thermopalmarium, Bacillus coagulans and Bacillus
thermoamylovorans were the culturable species at the desired operational temperature of
65°C. C. thermopalmarium and B. coagulans are major H2 producers with theoretical values
of 4mol H2/ g glucose and 3mol H2/ mol sugar, respectively. Sewage sludge is the best source
of biohydrogen producing bacteria in comparison to cow dung as an inoculum in AFBRs.
Bioreactor 1 with a constant operational temperature of 65°C and at a HRT of 6.5 allowed for
the desired Clostridium sp. to be the predominant H2 producer.
Date January 2012
CreatorsSebola, Keneilwe Mmule
Source SetsSouth African National ETD Portal
Detected LanguageEnglish
Formatapplication/pdf, application/pdf

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