Discussion

The Winogradsky Column was invented by Sergei Winogradsky in the 1880s as a simple
device by which a large diversity of Microorganisms can be cultured easily. The device consists
of a Transparent Column filled with Mud/Soil Sample as a solid medium for microbial growth
that is mixed with water, and a volume of air left in the column. The column is then covered to
prevent the evaporation of water and also the new addition of air, this maintains limited but equal
conditions for each organism, and enriched with nutrient sources to fit the requirements of the
variety of microorganisms needed to be cultured. The column is then incubated in light or dark
for a period of days in which large amounts of microbial growth will take place using the
provided nutrients. Different colonies of Bacteria Thrive at different areas of the column because
each area has a differing composition of air and nutrients as well as microbes thrive away from
areas of unfavourable conditions.

The Top layer of the column in the air space is strictly Oxic and provides oxygen for the
oxygenic microbes in the pond water below. Upon descending the column O2 Levels drop
significantly to close to Zero. Oxygen Levels become Microaerophillic in the water as well as the
top region of the soil in the column. Over a period of weeks motile oxygenic phototrophs thrive
in the aquatic pond water environment as the water provides medium for free movement, Algae
also thrives here. These Microorganisms are also green in colour showing the presence of a
photosynthetic mechanism which is why they proliferate at the top as light passes freely through
the water as well as they obtain more oxygen to engage in cellular respiration. Since
photosynthesis is taking place here, this creates an O2 Gradient where high O2 levels are at the
top of the column, and low O2 levels at the bottom. At the soil surface, green and blue green
microbes are also seen indicating the presence of Cyanobacteria Species.

At the bottom of the column conditions are strictly Anaerobic/Anoxic, CO2 levels are high
here this causes the proliferation of Sulphate Reducing Microbes like Desulfovibrio sp and
Clostridium sp which are Sulfidogenic meaning they breathe Sulphate instead of Molecular O2
as the final electron acceptor in respiration. This produces H2S in the soil which reacts with
Metal ions in the soil to form Metal Sulphate decompositions which gives the soil its black
sludgy colour. This in turn creates a sulphur gradient where high SO4 2- concentrations are at the
bottom of the column and low levels at the top. Above the Sulphate Reducers, H2S feeds into the
metabolism of 2 types of Photosynthetic Bacterium, Green Sulphur Bacterium e.g. Chlorobium
sp and above that, a layer of Purple Sulphur Bacterium e.g. Chromatium sp. These bacterium
uses CO2 as a carbon source, H2S as an electron donor and light as an energy source to produce
organic materials. Green Sulphur bacterium can tolerate higher levels of H2S that the Purple
Bacterium and thus purple Sulphur Bacterium proliferate in a region further away from the
Sulphate reducers. At the middle of the column and above, purple non sulphur bacterium thrive
as sulphur levels become very low going up the column e.g. Rhodomicrobium sp. These
bacterium use light for photosynthesis and use ethanol as an electron donor rather than H2S.
 When Sulphate levels rise in the aquatic environment, a unique microbial Genera of
filamentous bacteria tend to proliferate here i.e. Beggiotoa sp. As a method of survival, these
bacterium use H2S as an energy source and oxidizes it to sulphuric acid which uses this energy
as an energy source to fix carbon to produce organic molecules regulating the sulphate levels
here thus maintaining the sulphate gradient.

In this Experiment after a 4 week incubation period of the light column, algal growth at the
surface of the pond water as well as on the grass cutting in which it obtained its carbon from the
breakdown of the cellulose n the cell walls of the grass. Thick blue green Filamentous Growth
was seen at the top soil surface of the column and some green growth was seen a little above the
centre of the soil possible green sulphur bacterium species. Over the 4 weeks period Sulphate
reducers grew marvellously in abundance as a lot of black colouration was deposited in the soil
to more than half of the column to where the small speckles of possibly green sulphur bacterium
were. No growth of any other pigmented bacterium was observed from the column probably as a
result of the over accumulation of sulphate reducers increasing the H2S concentration in the soil
thus making the environment less favourable for bacterium such the purple sulphur bacterium
and non-sulphur bacterium to grow. Despite the observations from the column some rust red
coccoid bacterium were seen in the wet mount which could possibly be slowly growing purple
sulphur/non sulphur as a result of the unfavourable environment caused by the other microbes as
well as the usage up of nutrients and the oxygen availability. A lot of cyanobacteria were seen
under the wet mounts. The wet mounts of the dark column also showed microbes resembling that
off cyanobacteria and other photosynthetic organisms but their pigmentation was off probably
because of the face that they are trying to grown in an unfavourable environment and using other
metabolic pathways to sustain life.