| Summary: | Magister Scientiae - MSc (Biotechnology) === There has been a significant increase in the use of constructed wetlands globally for the
bioremediation of wastewater (Welz et al., 2011). In South Africa, the wine industry
generates more than one billion litres of wastewater annually and this is disposed of by
irrigation (Burton et al., 2007). A more cost effective and simple system need to be
employed for the treatment of winery effluent and a possible solution would be the use
of biological sand filters (BSFs). The microbial communities present in these BSFs play
an important role in the biodegradation of the organic wastewater pollutants.
Physicochemical and microbiological studies have been used to determine the
performance of BSFs for the remediation of winery effluent since 2009.
In this study, changes in the bacterial and fungal communities in different spatial niches
was analysed through the use of molecular fingerprinting techniques [terminal restriction
fragment length polymorphism (T-RFLP)] of the 16S rRNA gene and fungal ITS gene as
a response to the exposure to synthetic winery wastewater and to perform functional
studies using q-PCR on selected genes. Changes in the bacterial and fungal community
profiles were observed at the different niches after amendment with synthetic winery
wastewater. This result was confirmed by performing the phylogenetic analysis on the
bacterial population present within the BSF systems. Sand organisms including
Clostridium, Sarcina, Streptomyces, Actinobacteria were among the expected species
present in the sand samples at the deep inlet of the BSFs. Through the study by Burton
et al, (2007), one could hypothesise which organisms mentioned above were able to
metabolise the components of the synthetic winery wastewater to secondary
metabolites. Amplification of the functional gene through the use of qPCR of catechol
2,3-dioxygenase was successful. Increase in the amount of copy numbers between the
samples showed that the increase in expression of the catechol 2,3-dioxygenase meant
that there was an increase in the amount of organisms degrading the catehol build-up
in the BSF systems.
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