Microbial diversity and Molecular Biomonitoring for Efficient Treatment of Tannery Wastewater: Laboratory and Field- Scale Studies
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Date
2013-11
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Addis Ababa Universty
Abstract
This dissertation reports on culture- dependent and culture- independent characterization of
microorganisms of from tannery wastewater, with special reference to degradation of selected
retanning chemicals, In the culture- based study, four bacterial strains isolated ll'om a temporary
storage site of composite tanncry wastewater were reported to grow and utilizc the retanning
agcnts named as Basyantan AN and Retanal MD80 as their carbon and nitrogen sources. Ultra
Performance Liquid Chromatography (UPLC) analysis of compounds in the retanning agents
from batch cultures over a period of 14 days revealed an average of 18, 26 and 33% degradntion
of the three detected peaks of Basyantan by isolate B45. The same isolate also showed an
average of 34% degradation of Retana!' Kinetic studies conducted on this isolnte showed the
dependence of growth on the concentration of Retanal as a substrate, with a maximum grO\\1h
rate (Jlma,)of 1450 cells mr'.hr" and a half saturation constant (Ks) of 10.16 mg.r'. On the basis
of 16S rRNA gene amplification, sequencing and phylogenetic analysis, the four isolates werc
identified as Micrococcus luleus strain B lR, Brevibacleriulll pilyocalllpae strain B4500R,
Lysinibacillus sphaericlls strain R 1902R and Enlerococcus gallina/'lllJl Strain R280 I R.
Bench- scale study involving a sequencing batch reactor (SBR) run for 42 days for studying the
biodegradation of Basyantan and Retanal revealed a pronounced degradation up to 100%
regarding peak 2 of Basyantan. Dynamics of sludge microbial sub- communities as followed by
T-RFLP showed the dominance of membcrs of the class Burkholderaceae and a less diverse
community in the first five batches of the SBR run; which was then succeeded by members
belonging to the phylum Bacteroidetes and classes Rhodocycalaceae, Rhodobacteraceae,
Pseudomonadaceae and Bradyrhizobiaceae; with increased diversity towards the end of the run.
Flow cytometric analysis of bacterial cells in the sludge was used to create pattcrns mirroring
dynamics of bacterial community over the period of SBR run. The patterns were resolved in 30
sub, clusters eight of which wcre sOlicd and members identified. Correlation of the bacterial
abundancc in thc sortcd sub- clusters with the measured abiotic data showed the prcsence of a
strong (p< 0.0 I) positive correlation bctween degradation of Basyantan and Retanal with
mcmbers of Bacteroidetes and Protcobactcria, indicating the key degradation role played by
thesc members.
Ficld- scale study on the structure and diversity of the microbial community in the sludge of the
Anaerobic- aerobic reactors as well as the root area of a constructed wetland treating tannery
effluent located in Modjo tannery was conducted using the 16S ribosomal RNA gene scquencing
performcd for cach of the reactors. The results revealed a total of 31 different phylotypes with
the dominant members belonging to FirllliclIIes, Bacleroidia, Proleobacleria, Sphingobacleria
and S)'Ilergislia in the anaerobic and aerobic reactors. In the constructcd wetland, members of
thc group C)'({l1obaCleria were also identified in addition to Firmicules, 13acleroidia and
Proleobaclel'ia. Thc presence of members of groups that arc associated with reduction of sulfate and removal of chromium, nitrate and degradation of synthetic aromatic compounds indicates
their implication in the current performance of the treatment system. Furthermore, physicochemical
data (total Nitrogen, total Carbon, pH, Sulphate, Nitrate) have been recorded in the
anaerobic, aerobic and constructed wetlands and analyses of these data elucidate the key factor
structuring the observed microbial communities.
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Keywords
Laboratory and Field- Scale Studies