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Microscopic visualisation of succinate producing biofilms of actinobacillus succinogenes

Dissertation (MEng (Chemical Engineering))--University of Pretoria, 2017.

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Other Authors: Nicol, Willie
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Published: University of Pretoria 2017
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access_status_str Open Access
author2 Nicol, Willie
author_browse Nicol, Willie
author_facet Nicol, Willie
collection Thesis
dc_rights_str_mv © 2017 University of Pretoria. All rights reserved. The copyright in this work vests in the University of Pretoria. No part of this work may be reproduced or transmitted in any form or by any means, without the prior written permission of the University of Pretoria.
description Dissertation (MEng (Chemical Engineering))--University of Pretoria, 2017.
format Thesis
id oai:repository.up.ac.za:2263/62782
institution University of Pretoria (South Africa)
last_indexed 2026-06-10T12:38:15.015Z
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provenance_str_mv Harvested via OAI-PMH from UPSpace — University of Pretoria Institutional Repository
publishDate 2017
publishDateRange 2017
publishDateSort 2017
publisher University of Pretoria
publisherStr University of Pretoria
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source_str UPSpace — University of Pretoria Institutional Repository
spelling oai:repository.up.ac.za:2263/62782 Microscopic visualisation of succinate producing biofilms of actinobacillus succinogenes Nicol, Willie u11119072@tuks.co.za Mokwatlo, Sekgetho Charles Actinobacillus succinogenes Biofilm structure Scanning electron microscopy Confocal scanning laser microscopy UCTD Engineering, built environment and information technology theses SDG-09 Engineering, built environment and information technology theses SDG-12 Dissertation (MEng (Chemical Engineering))--University of Pretoria, 2017. Biofilms of Actinobacillus succinogenes, grown in a biofilm reactor system, were investigated for structure and cell viability, through microscopic visualisation with a confocal scanning laser microscope (CSLM) and a scanning electron microscope (SEM). Biofilms were sampled and visualised at steady state conditions with the broth containing succinic acid titres between 15 and 21 g/L. All sampled biofilm was 6 days old. Six-day-old biofilms of A. succinogenes showed a heterogeneous biofilm architecture composed of cell micro-colony pillars which varied considerably in thickness, area and shape. Microcolony pillars consisted of a densely packed entanglement of sessile cells. Quantitative analysis revealed that the pillars were mostly large, with a mean pillar diameter of 170 m and a mean thickness of 92 m, although pillar diameter and thickness were variable as they ranged from 25 – 500 m and 30 – 300 m, respectively. In the regions close to the substratum surface, pillars were characterised by having defined borders with a network of channels ranging from 40 – 200 m in width separating them. However, towards the middle of the biofilm depth some of the pillars coalesced. For this reason low cross sectional area coverage of biofilm consistently occurred at the bottom portion of the biofilm whilst the highest coverage was in the middle portion of the biofilm. Regarding cell morphology, very large differences were observed. Planktonic cells were rod-shaped, whereas sessile cells expressed an elongated rod morphology and thus were much longer and thinner compared with planktonic cells. Planktonic cells were 1 – 2 m thick and 4 – 5 m long, while sessile cells were 0.5 – 1 m thick and 5 – 100 m long. Long sessile cells resulted in extensive tangling in microcolony pillars, which may have contributed to the structural stability of the pillars. Fibre-like connections of constant diameter were observed between cells, and between the cells and surface. The diameter of these connections was approximately 20 – 30 nm. Viability stains showed that in the bottom portion (from 0 - 20 m above the substratum surface) of the biofilm, most of the cells were dead. However, the portion of covered area attributed to living cells increased past the middle of the biofilm towards the top part of the biofilm. A high percentage of living cells was thus found towards the top part of the biofilm. Overall, 65% (with 2% standard deviation) of the entire biofilm was composed of dead cells. In this way, the results show that operation at high acid conditions comes at a cost of low overall biomass productivity due to decreased active biomass. mi2026 Chemical Engineering MEng (Chemical Engineering) Unrestricted SDG-09: Industry, innovation and infrastructure SDG-12: Responsible consumption and production 2017-10-13T13:41:20Z 2017-10-13T13:41:20Z 2017-09-08 2017 Dissertation Mokwatlo, SC 2017, Microscopic visualisation of succinate producing biofilms of Actinobacillus succinogenes, MEng Dissertation, University of Pretoria, Pretoria, viewed yymmdd <http://hdl.handle.net/2263/62782> S2017 http://hdl.handle.net/2263/62782 © 2017 University of Pretoria. All rights reserved. The copyright in this work vests in the University of Pretoria. No part of this work may be reproduced or transmitted in any form or by any means, without the prior written permission of the University of Pretoria. application/pdf University of Pretoria
spellingShingle Actinobacillus succinogenes
Biofilm structure
Scanning electron microscopy
Confocal scanning laser microscopy
UCTD
Engineering, built environment and information technology theses SDG-09
Engineering, built environment and information technology theses SDG-12
Microscopic visualisation of succinate producing biofilms of actinobacillus succinogenes
title Microscopic visualisation of succinate producing biofilms of actinobacillus succinogenes
title_full Microscopic visualisation of succinate producing biofilms of actinobacillus succinogenes
title_fullStr Microscopic visualisation of succinate producing biofilms of actinobacillus succinogenes
title_full_unstemmed Microscopic visualisation of succinate producing biofilms of actinobacillus succinogenes
title_short Microscopic visualisation of succinate producing biofilms of actinobacillus succinogenes
title_sort microscopic visualisation of succinate producing biofilms of actinobacillus succinogenes
topic Actinobacillus succinogenes
Biofilm structure
Scanning electron microscopy
Confocal scanning laser microscopy
UCTD
Engineering, built environment and information technology theses SDG-09
Engineering, built environment and information technology theses SDG-12
url http://hdl.handle.net/2263/62782