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Characterization of Bacterial Biofilms for Wastewater Treatment
KTH, School of Biotechnology (BIO), Environmental Microbiology. (Miljömikrobiologi)
2009 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Research performed at the Division of Environmental Microbiology has over the last years resulted in the isolation of possible bacterial key-organisms with efficient nutrient removal properties (Comamonas denitrificans, Brachymonas denitrificans, Aeromonas hydrophila). Effective use of these organisms for enhanced nutrient removal in wastewater treatment applications requires the strains to be retained, to proliferate and to maintain biological activity within theprocess. This can be achieved by immobilization of the organisms using an appropriate system.Two putative immobilization systems, agar entrapment and biofilm formation, wereassessed. Surface attached biofilm growth provided better results with respect to cell retention,proliferation and microbial activity than immobilization in agar beads. Thus, biofilm physiology was further characterized using simplified systems of single, dual or multi strain bacterial consortia containing the key-organisms as well as other wastewater treatment isolates. Mechanisms for initial adherence, biofilm formation over time, dynamics and characteristics of extracellular polymeric substances (EPS) and exopolysaccharides, nutrient removal activity as well as the effect of bacterial interactions were investigated. The results showed that all theassessed bacterial strains could form single strain biofilm providing that a suitable nutrientsupply was given. Production of EPS was found to be critical for biofilm development and both EPS and polysaccharide residue composition varied with bacterial strain, culture conditions and biofilm age. Denitrification and phosphorus removal activity of the keyorganisms was maintained in biofilm growth. Co-culturing of two or more strains resulted in both synergistic and antagonistic effects on biofilm formation as well as the microbial activitywithin the biofilm. Bacterial interactions also induced the synthesis of new polysaccharideswhich were not produced in pure strain biofilms.The complexity of single and mixed strain biofilm development and the implications of interactions on biofilm performance were underlined in this study. The data presented can be useful for modeling of biofilm systems, serve as a tool for selection of bacterial strain combinations to use for bioaugmentation/bioremediation or provide a base for further experiment design.

Place, publisher, year, edition, pages
Stockholm: Kungliga Tekniska Högskolan , 2009. , viii, 63 p.
Series
Trita-BIO-Report, ISSN 1654-2312
Keyword [en]
biofilm, extracellular polymeric substances, exopolysaccharides, interspecies interactions, wastewater treatment, denitrification, phosphorus removal
National Category
Microbiology
Identifiers
URN: urn:nbn:se:kth:diva-10118ISBN: 978-91-7415-255-5 (print)OAI: oai:DiVA.org:kth-10118DiVA: diva2:209486
Public defence
2009-04-17, Hörsal K1, KTH, Teknikringen 56, Stockholm, 10:00 (Swedish)
Opponent
Supervisors
Note
QC 20100622Available from: 2009-03-25 Created: 2009-03-23 Last updated: 2010-07-19Bibliographically approved
List of papers
1. Bioaugmentation for enhanced denitrification in a labscale treatment system
Open this publication in new window or tab >>Bioaugmentation for enhanced denitrification in a labscale treatment system
2006 (English)In: Proceedings of the Second IASTED International Conference on Advanced Technology in the Environmental Field / [ed] Ubertini, L, ACTA Press, 2006, 63-67 p.Conference paper, Published paper (Refereed)
Abstract [en]

The effect of bioaugmentation was investigated in a predenitrification lab-scale wastewater treatment system. The aim was to investigate the difference between two approaches to bioaugmentation: one in which suspended overnight culture was used as inoculum and another where bacteria immobilized in 1% agar beads were used. Pure cultures of the denitrifying bacteria Comamonas denitrificans ATCC 700936T were used in the experiments. The effect of bioaugmentation on the system was monitored using fluorescent in situ hybridization (FISH) and denitrification activity tests. The bioaugmentation with suspended bacteria showed a rapid initial (4 days) increase in denitrification activity. After 8 days the activity declined to the level of the reference system and cells of C. denitrificans were no longer detectable. Augmentation with agar-embedded bacteria resulted in a small increase in activity and very few bacteria of C. denitrificans could be observed.

Place, publisher, year, edition, pages
ACTA Press, 2006
Keyword
Bioaugmentation, denitrification, fluorescent in situ hybridization, immobilization, wastewater treatment, Comamonas denitrificans
National Category
Environmental Sciences
Identifiers
urn:nbn:se:kth:diva-10152 (URN)000236368300012 ()2-s2.0-38049140883 (Scopus ID)978-088986554-9 (ISBN)
Conference
2nd IASTED International Conference on Advanced Technology in the Environmental Field Location: Lanzarote, Spain, Date: FEB 06-08, 2006
Note

QC 20100622

Available from: 2009-03-25 Created: 2009-03-25 Last updated: 2014-11-07Bibliographically approved
2. Biofilm formation and interactions of bacterial strains found in wastewater treatment systems
Open this publication in new window or tab >>Biofilm formation and interactions of bacterial strains found in wastewater treatment systems
2008 (English)In: FEMS Microbiology Letters, ISSN 0378-1097, E-ISSN 1574-6968, Vol. 283, no 1, 83-90 p.Article in journal (Refereed) Published
Abstract [en]

Biofilm formation and adherence properties of 13 bacterial strains commonly found in wastewater treatment systems were studied in pure and mixed cultures using a crystal violet microtiter plate assay. Four different culture media were used, wastewater, acetate medium, glucose medium and diluted nutrient broth. The medium composition strongly affected biofilm formation. All strains were able to form pure culture biofilms within 24 h in at least one of the tested culture media and three strains were able to form biofilm in all four culture media, namely Acinetobacter calcoaceticus ATCC 23055, Comamonas denitrificans 123 and Pseudomonas aeruginosa MBL 0199. The adherence properties assessed were initial adherence, cell surface hydrophobicity, and production of amyloid fibers and extracellular polymeric substances. The growth of dual-strain biofilms showed that five organisms formed biofilm with all 13 strains while seven formed no or only weak biofilm when cocultured. In dual-strain cultures, strains with different properties were able to complement each other, giving synergistic effects. Strongest biofilm formation was observed when a mixture of all 13 bacteria were grown together. These results on attachment and biofilm formation can serve as a tool for the design of tailored systems for the degradation of municipal and industrial wastewater.

Place, publisher, year, edition, pages
OXFORD: BLACKWELL PUBLISHING, 2008
Keyword
biofilm formation; FISH; initial adherence; wastewater treatment
National Category
Industrial Biotechnology
Identifiers
urn:nbn:se:kth:diva-10153 (URN)10.1111/j.1574-6968.2008.01149.x (DOI)000255286100013 ()2-s2.0-42649086030 (Scopus ID)
Note
QC 20100622Available from: 2009-03-25 Created: 2009-03-25 Last updated: 2010-12-03Bibliographically approved
3. Assessment of carrier materials for biofilm formation and denitrification
Open this publication in new window or tab >>Assessment of carrier materials for biofilm formation and denitrification
2008 (English)In: Vatten, ISSN 0042-2886, Vol. 64, 201-207 p.Article in journal (Refereed) Published
Place, publisher, year, edition, pages
Lund: Föreningen vatten, 2008
National Category
Industrial Biotechnology
Identifiers
urn:nbn:se:kth:diva-10154 (URN)
Note
QC 20100622Available from: 2009-03-25 Created: 2009-03-25 Last updated: 2010-07-19Bibliographically approved
4. Characterization of extracellular polymeric substances from denitrifying organism Comamonas denitrificans
Open this publication in new window or tab >>Characterization of extracellular polymeric substances from denitrifying organism Comamonas denitrificans
2009 (English)In: Applied Microbiology and Biotechnology, ISSN 0175-7598, E-ISSN 1432-0614, Vol. 82, no 3, 535-543 p.Article in journal (Refereed) Published
Abstract [en]

Extracellular polymeric substances (EPS) play an important role in the formation and activity of biofilms in wastewater treatment (WWT). The EPS of the denitrifying biomarker Comamonas denitrificans strain 110, produced in different culture media and growth modes, were characterized. The EPS mainly contained protein (3-37%), nucleic acids (9-50%), and carbohydrates (3-21%). The extracellular DNA was found to be important for initial biofilm formation since biofilm, but not planktonic growth, was inhibited in the presence of DNase. The polysaccharide fraction appeared to consist of at least two distinct polymers, one branched fraction (A) made up of glucose and mannose with a molecular weight around 100 kDa. The other fraction (B) was larger and consisted of ribose, mannose, glucose, rhamnose, arabinose, galactose, and N-acetylglucosamine. Fraction B polysaccharides were mainly found in capsular EPS which was the dominant type in biofilms and agar-grown colonies. Fraction A was abundant in the released EPS, the dominant type in planktonic cultures. Biofilm and agar-grown EPS displayed similar overall properties while planktonic EPS showed clear compositional disparity. This study presents results on the physiology of a key WWT organism, which may be useful in the future development of improved biofilm techniques for WWT purposes.

Keyword
Biofilm; Comamonas denitrificans; GC-MS; Extracellular polymeric substances; HPAEC; Polysaccharides
National Category
Industrial Biotechnology
Identifiers
urn:nbn:se:kth:diva-10155 (URN)10.1007/s00253-008-1817-3 (DOI)000263484600014 ()2-s2.0-60549106192 (Scopus ID)
Note
QC 20100622Available from: 2009-03-25 Created: 2009-03-25 Last updated: 2010-12-03Bibliographically approved
5. Biological nutrient removal by individual and mixed strain biofilms
Open this publication in new window or tab >>Biological nutrient removal by individual and mixed strain biofilms
(English)Manuscript (Other academic)
National Category
Industrial Biotechnology
Identifiers
urn:nbn:se:kth:diva-10157 (URN)
Note
QC 20100622Available from: 2009-03-26 Created: 2009-03-26 Last updated: 2010-07-19Bibliographically approved
6. Persistence and competition of denitrifying biofilms subjected to a natural wastewater flora
Open this publication in new window or tab >>Persistence and competition of denitrifying biofilms subjected to a natural wastewater flora
(English)Manuscript (Other academic)
National Category
Industrial Biotechnology
Identifiers
urn:nbn:se:kth:diva-10158 (URN)
Note
QC 20100622Available from: 2009-03-26 Created: 2009-03-26 Last updated: 2010-07-19Bibliographically approved

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