Abstract:
The major goal of microbial ecology is to understand microbial diversity in natural
habitats their interaction with one another and with their habitat. The soda lakes are
highly productive environments and the soda lakes of the East African Rift valley
have been shown to support a dense and diverse population of aerobic,
organotrophic, halophilic, alkaliphilic and alkalitolerant representatives of major
bacterial and archaeal phyla.
The isolation and characterization of organisms belonging to widespread but
previously uncultivated groups of organisms can provide insights into the roles and
functions of these organisms in their natural settings and assist in the formulation
of hypotheses about metabolic interactions between microorganisms and their
natural environment. Several studies have been carried out to document the
microbial diversity of the Kenyan soda lakes by other researchers. However no
comprehensive study has been done in Lake Elmenteita. The aim of this study was
to assess the microbial diversity of Lake Elmenteita using both culture independent
and culture dependent techniques. The application of both techniques was expected
to provide new insights into the microbial diversity of the Lake as well as possible
roles played by each group within the soda lake environment.
Application of molecular tools to study microbial ecology has widened our
approximation of diversity in the environments. Clone Libraries were constructed
from PCR amplicons from total environmental DNA. Primers specific for Bacteria
and Archaea respectively were used. Partial sequences were generated for both the
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clones and the isolates. The relatedness of the Lake Elmenteita bacterial rRNA
sequences to known rRNA gene sequences was determined by BLAST analysis
and by alignment to the sequences on the ARB database (Release, 1994).
Clones possessed a higher similarity to other environmental clones than to cultured
microorganisms. A total of 655 clone sequences were sequenced. Of these 525
(80.15%) sequences were related to uncultured members of the Domain Bacteria.
This indicates that a large proportion of deep phylogenetic groups are represented
in the clone libraries. Sixteen percent of the clones had similarity values below
90% to both cultured and uncultured microorganisms. Forty three percent of the
clones had similarity values between 90-95% as compared to 34.35% that had
values between 96-98%. Only a mere 6.87% had values between 99-100%.
However a number of factors including relatively low cell numbers of large
organisms and a variable number of rRNA operons among organisms, as well as
extraction and PCR bias, may lead to under-representation of phylotypes relative to
their in situ abundance.
Cultured isolates are still very important in developing our understanding of
bacterial physiology, genetics, and ecology. Isolation was done using both nutrient
rich and nutrient poor media. A polyphasic approach was employed in the
identification of the various strains. The majority of the isolates (36.75%) belonged
to the genus Halomonas while 31.35% belonged to the Genus Bacillus. More than
half of the isolates (59.45%) belonged to the Gammaproteobacteria. An overlap
between the clone library and the isolates was observed in the Order Bacillales and
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the Actinobacteria only. In this study novel isolates related to Marinospirillum,
Idiomarina, Streptomyces, ocardia, Marinilactibacillus, Amphibacillus and
Vibrio were recovered. A polyphasic approach to characterization showed they
represented novel taxa.
The study showed that the application of both culture dependent and culture
independent methods gives a better picture of diversity in the environment. It can
be concluded the soda lakes harbour novel uncultured groups of microorganisms
and most of them are of biotechnological potential. Future work should focus on
Archaeal diversity as well as the uncultured groups of bacteria.