Thesis submitted in fulfilment of the requirements for the degree
Doctor of Technology: Biomedical Technology
in the Faculty of Health and Wellness Sciences
at the Cape Peninsula University of Technology
2008 / River systems can become contaminated with micro-organisms and metals and the
routine monitoring of these rivers is essential to control the occurrence of these
contaminants in water bodies. This study was aimed at investigating the metal
contamination levels in the Berg-, Plankenburg- and Diep Rivers in the Western Cape,
South Africa, followed by the remediation of these rivers, using bioreactor systems.
Sampling sites were identified and samples [water, sediment and biofilm (leaves,
rocks and glass, etc.)] were collected along the Berg- and Plankenburg Rivers from May
2004 to May 2005 and for the Diep River, from February 2005 to November 2005. The
concentrations of aluminium (Al), copper (Cu), iron (Fe), manganese (Mn), nickel (Ni),
lead (Pb) and zinc (Zn) were determined using the nitric acid digestion method and
analysed by Inductively Coupled Plasma Atomic Emission Spectrometry (ICP-AES).
For the Berg River, the highest concentrations in water samples were recorded
for Al, Mn and Fe at the agricultural area (Site A – chapter 2). In the sediment and
biofilm samples, the highest metal concentrations were once again recorded for Al and
Fe. The concentrations of Al and Fe were significantly higher (p < 0.05) than than Cu,
Zn, Pb, Ni and Mn in water, sediment and biofilm samples, and were mostly higher than
the quality guidelines recommended by the Department of Water Affairs and Forestry
(DWAF, 1996) and the Canadian Council for the Ministers of the Environment (CCME,
2001). Possible sources of contamination in the Berg River could be due to the leaching
or improper discarding of household waste from the informal- and established residential
areas, as well as the improper discarding of pesticides at the agricultural area.
For both the Plankenburg and Diep Rivers the Al and Fe concentrations were
higher than all the other metals analysed for in sediment and water samples. The
highest concentrations recorded in the Plankenburg River was 13.6 mg.l-1 (water - Week
18, Site B) and 15 018 mg.kg-1 (sediment - Week 1, Site C) for Al and 48 mg.l-1 (water -
Week 43, Site A) and 14 363.8 mg.kg-1 (sediment - Week 1, Site A) for Fe. The highest
concentrations recorded in the Diep River was 4 mg.l-1 (water - Week 1, Site A) and
19 179 mg.kg-1 (sediment - Week 1, Site C) for Al and 513 mg.l-1 (water - Week 27, Site
A) and 106 379.5 mg.kg-1 (sediment - Week 9, Site C) for Fe. For most of the metals
analysed the concentrations were higher than the recommended water quality
guidelines as stipulated by the Department of Water Affairs and Forestry (DWAF,
1996b), the Canadian Council for the Ministers of the Environment (CCME, 2001) and
the ‘World average’ (Martin and Windom, 1991). Point sources of pollution could not
conclusively be identified, but the industrial and residential areas could have influenced
the increased concentrations. Metal concentrations should be routinely monitored and
the guidelines should be updated and revised based on the current state of the rivers
and pollution influences.
Micro-organisms isolated from flow cells after exposure to varying metal
concentrations were investigated for possible metal-tolerance. A site where high metal
concentrations were recorded along the Plankenburg River was investigated. The
micro-organisms isolated from the flow cells were cultured and identified using the
Polymerase Chain Reaction (PCR) technique, in conjunction with universal 16SrRNA
primers. The phylogeny of the representative organisms in GenBank, were analysed
using the Neighbour-joining algorithm in Clustal X. After exposure, the channels were
stained with the LIVE/DEAD BacLightTM viability probe and visualised using
Epifluorescence Microscopy. The results revealed that when exposed to the highest
concentrations of Al (900 mg.l-1), Fe (1000 mg.l-1), Cu (10 mg.l-1) and Mn (80 mg.l-1), the
percentage of dead cells increased, and when exposed to the lowest concentrations of
Al (10 mg.l-1), Cu (0.5 mg.l-1), Mn (1.5 mg.l-1) and Zn (0.5 mg.l-1), no significant
differences could be distinguished between live an dead cells. When exposed to the
highest concentrations of Zn (40 mg.l-1) and Ni (20 mg.l-1), no significant differences
between the live and dead cell percentages, were observed. The phylogenetic tree
showed that a diverse group of organisms were isolated from the flow cells and that
some of the isolates exhibited multiple metal resistance (Stenotrophomonas maltophilia
strain 776, Bacillus sp. ZH6, Staphylococcus sp. MOLA:313, Pseudomonas sp. and
Delftia tsuruhatensis strain A90 exhibited tolerance to Zn, Ni, Cu, Al, Fe), while other
isolates were resistant to specific metals (Comamonas testosteroni WDL7,
Microbacterium sp. PAO-12 and Sphingomonas sp. 8b-1 exhibited tolerance to Cu, Ni
and Zn, respectively, while Kocuria kristinae strain 6J-5b and Micrococcus sp. TPR14
exhibited tolerance to Mn).
The efficiency of two laboratory-scale and one on-site bioreactor system was
evaluated to determine their ability to reduce metal concentrations in river water
samples. The laboratory-scale bioreactors were run for a two-week and a three-week
period and the on-site bioreactor for a period of ten weeks. Water (all three bioreactors)
and bioballs (bioreactor two and on-site bioreactor) were collected, digested with 55%
nitric acid and analysed using ICP-AES. The final concentrations for Al, Ni and Zn
(bioreactor one) and Mn (bioreactor two), decreased to below their recommended
concentrations in water samples. In the on-site, six-tank bioreactor system, the
concentrations for Fe, Cu, Mn and Ni decreased, but still exceeded the recommended
concentrations. The concentrations recorded in the biofilm suspensions removed from
the bioballs collected from bioreactor two and the on-site bioreactor, revealed
concentrations higher than those recorded in the corresponding water samples for all
the metals analysed, except Fe. The bioballs were shown to be efficient for biofilm
attachment and subsequent metal accumulation. The species diversity of the organisms
isolated from the bioreactor (bioreactor two) experiment after three days (initial) differed
from the organisms isolated after 15 days (final). Hydrogenophaga sp., Ochrobactrum
sp, Corynebacterium sp., Chelatobater sp. and Brevundimonas sp. were present only at
the start of the bioreactor experiment. The surviving populations present both in the
beginning and at the end of the bioreactor experiment belonged predominantly to the
genera, Pseudomonas and Bacillus. Metal-tolerant organisms, such as Bacillus,
Pseudomonas, Micrococcus and Stenotrophomonas, amongst others, could possibly be
utilised to increase the efficiency of the bioreactors. The bioreactor system should
however, be optimised further to improve its efficacy.
| Identifer | oai:union.ndltd.org:netd.ac.za/oai:union.ndltd.org:cput/oai:localhost:20.500.11838/1526 |
| Date | January 2008 |
| Creators | Jackson, Vanessa Angela |
| Publisher | Cape Peninsula University of Technology |
| Source Sets | South African National ETD Portal |
| Language | English |
| Detected Language | English |
| Type | Thesis |
| Rights | http://creativecommons.org/licenses/by-nc-sa/3.0/za/ |
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