MASMANIDOU MARIA

ZACHARIADI ISAVELLA
UNIVERSITY OF IOANNINA GREECE
BIOLOGICAL APLICATIONS AND TECHNOLOGY
AQUATIC MICROORGANISMS

WEEK: 2nd

LEFT: ¾ sediment-enrichment materials and lake water

RIGHT: ¼ sediment-enrichment materials, 2/4 sediment and lake water

On the left column, we observed that the aquatic phase got a dark green colour because of the growth of  green sulpur bacteria, such as chlorobium. Those microorganisms gain energy from light reactions and produce their cellular materials from CO2 in much the same way as plants do. However, there is one essential difference, they do not generate oxygen during photosynthesis because they do not use water as the reductant, instead, they use H₂S. We also noticed a black line on the sediment’s surface which provides the existence of black sulphur-reducing bacteria such as clostridium and desulfovibrio. They use sulphate or other partly oxidised forms of sulphur as the terminal electron acceptor, generating large amounts of H₂S by this process. The H₂S react with any iron in the sediment, producing black ferrous sulphide. This is why lake sediments are frequently black. However, some of the H₂S diffuses upwards into the water column, where it is utilised by other organisms (e.g green sulpur bacteria).

On the other hand, on the right column, we observed just a light green colour in the aquatic phase which could be a proof for the existence of Algae and Photosynthetic cyanobacteria(those microorganisms contains chlorophyll a and performs oxygenic photosynthesis).

From those indications we accepted the first hypothesis that microorganisms in the left bottle grew up faster than the right one.

WEEK: 3^rd – 4^rth

LEFT: ¾ sediment-enrichment materials and lake water

RIGHT: ¼ sediment-enrichment materials, 2/4 sediment and lake water

Our columns had no big difference between the 3rd and 4th week so we took just one photo.

The left turned purple  because of the growth of purple sulphur bacteria. These bacteria grow in anaerobic conditions, gaining their energy from light reactions but using organic acids as their carbon source for cellular synthesis. So they are termed photoheterotrophs. The organic acids that they use are the fermentation products of other anaerobic bacteria (e.g. Clostridium species). On the right one, aquatic phase turned dark green because of the green sulphur bacteria. No difference was observed in the sediment in both columns.

WEEK: 5^th

LEFT: ¾ sediment-enrichment materials and lake water

RIGHT: ¼ sediment-enrichment materials, 2/4 sediment and lake water

Both of our columns got darker and so swelled that they could not remain in an upright position. When we tried to open them, bubbles started to emerge, which is a proof of the existence and the increase in the amount of methanogens. Except the methanogens, there might have been some other obligatory anaerobic bacteria at the bottom, such as Clostridium or Desulfovibrio. Then, Desulfovibrio respire using these compounds to reduce the sulfate from the eggs. These processes quickly deplete any remaining of O₂ at the bottom of the column. Desulfovibrio release Hydrogen-Sulfide as a byproduct of said sulfate reduction. This causes a concentration gradient in the column between O₂ and H₂S (Higher O₂ at top). On the left bottle, in the aquatic anaerobic phase, grew up both green and purple photosynthetic sulphur bacteria. On the other hand,the right bottle got a purple-red colour because of the sulphur or non sulphur  photosynthetic bacteria (such as Rhodomicrobium). These bacteria use the Ethanol which produced from the clostridium as a photosynthetic reducer.

WEEK: 6^th

LEFT: ¾ sediment-enrichment materials and lake water

RIGHT: ¼ sediment-enrichment materials, 2/4 sediment and lake water

Six weeks later the column with the excess of enrichment materials (left one) got even darker in both sediment and aquatic phase. Microorganisms such as black sulphur-reducing  bacteria (in the sediment phase) and green sulphur bacteria (in the aquatic phase) continue growing up. In the right column, the aquatic phase got an orange-red colour because of the purple sulphur or non sulphur bacteria. Moreover, that colour could probably indicate cyanobacteria, who lost their chlorophylls and turned orange. No difference observed in the sediment phase in that column.

WEEK: 7^th

LEFT: ¾ sediment-enrichment materials and lake water

RIGHT: ¼ sediment-enrichment materials, 2/4 sediment and lake water

The last week, in the sediment phase, in both of our columns developed some white bacteria ( basically colourless bacteria), which use sulphate as the terminal electron acceptor, generating large amounts of H₂S by this process. Those microorganisms grow up in completely anaerobic conditions. Furthermore, both of our columns in aquatic phase turned green-orange because of the green and purple sulphur bacteria. Finally, in the left column,  grew up something like moss plants in the aquatic phase and biofilm in both columns.

Winogradsky’s Column

UNIVERSITY OF IOANNINA GREECE
BIOLOGICAL APLICATIONS AND TECHNOLOGY

AQUATIC MICROORGANISMS 

Team 7: Serasidis Konstantinos, Tsinoglou Makrina

Weeks: 5-7

During these three weeks there weren't observed any significant differences between the columns. However, between these weeks and the 4^th week a main difference has been noticed in the Control column. The color of the water in the column appears to have green- brown color. In contrary with the 4^th week the water had bright orange color. This difference is probably caused by the growth of cyanobacteria. Cyanoabcteria have oxygen-evolving photosynthesis like that of plants. Once the cyanobacteria start to grow they can oxygenate most of the water. So, the conditions in the water became aerobic and the purple non sulfur bacteria that need anaerobic conditions and are responsible for the orange color can’t grow anymore. That’s why the color in the water turned from orange to green after a few weeks.

The 7^th week is the last week of Winogradsky’s columns growth. The experiment ended successfully and our hypothesis was confirmed. Our hypothesis was that in conditions that light is absence the growth of phototrophic bacteria will be absence too. The results have showed that clearly, as green and orange color that indicates the growth of phototrophic bacteria was present in the Control column and absence in column in the dark.

2.Column in the dark 

1. Control column  

STUDENTS:NENA KOUKOUGELI
                        DIMITRA-IOLI SKOUROLIAKOU
UNIVERSITY OF IOANNINA GREECE
BIOLOGICAL APLICATIONS AND TECHNOLOGY
AQUATIC MICROORGANISMS

WEEK: 7th

LEFT: warmth-exposed

RIGHT: control

   Warmth – exposed column                                          Control column

  

  Between the 6^th and 7^th week, we couldn’t descry any obvious difference.

  The growth of the purple sulfur bacteria in the middle zone is still more visible in the control column than in the warmth-exposed column. During photosynthesis, unlike cyanobacteria or plants, they bind sulfur from the bottom of the column producing hydrogen sulfide, rather than oxygen. Thus, the purple sulfur bacteria cause anoxic conditions in the middle zone they grow.

  In both columns, is still observed the presence of biofilms, the concentration of which, is greater in the first column.

  This gradual distinction in the zones of the column, starting with the display of sulphates purple bacteria, has only been observed in the control column, even after 7 weeks. Thus, it can be assumed that the growth rate of the bacteria in the 2^nd column, is considerably slower, as zoning hasn’t been observed yet. This fact is probably due to the rapid fluctuation of temperature. In the warmth-exposed column, this affects the bacteria directly by limiting the growth of the non-resistant ones at high temperatures. Also, the bacteria in this column, in order to get protected from the temperature fluctuation, could form spores, which means a static metabolic condition. Rather than that, the overheating of the plastic bottle can excrete damaging chemicals for the bacteria.
  

Hypothesis: The rapid fluctuation of the temperature on daily basis, reduces the growth rate of the microorganisms in the 2^nd Winogradsky column

UNIVERSITY OF IOANNINA GREECE
BIOLOGICAL APLICATIONS AND TECHNOLOGY

AQUATIC MICROORGANISMS 

Team 7: Serasidis Konstantinos, Tsinoglou Makrina
Week: 4

Fourth week’s observation: 18/11/2016

Between the previous weeks and this week’s observations several changes have been noticed in the Control column. Firstly, at the bottom of the column green color has been noticed. Specifically, these changes state the growth of green sulfur bacteria. These bacteria require anaerobic conditions, sulfide ions and light effect in order to photosynthesize. Also, at the bottom of the column are noticeable black spots that are caused by anaerobic sulfur-reducing bacteria. These bacteria use either sulfate or oxidized forms of sulfur as the terminal electron acceptor, generating large amounts of H₂S. The H₂S will react with any iron in the sediment, producing black ferrous sulfide. At a higher level of the column a brown color in the sediment shows the growth of heterotrophic bacteria. There is a green color band immediately above the sediment. The green colored band is caused by the diffusion of H₂S from the sediment into the water column that enables anaerobic photosynthetic bacteria to grow. The orange color in the water of the column that is caused of purple non-sulfur bacteria seems to be the same as the previous week. Finally, biofilm is observed at the top of the water column. A biofilm is any group of microorganisms in which cells stick to each other and often these cells adhere to a surface.

At the bottom of the column in the dark, the sediment is observed darker in comparison with the previous week. The darker color of the sediment indicates the increasing growth of anaerobic bacteria. Also, black spots are noticed at the lower levels of the sediment that are caused by anaerobic sulfur-reducing bacteria. Additionally, the black band which formed at the upper levels of the sediment is caused by the growth anaerobic sulfur-reducing bacteria. The anaerobic conditions are formed by the diffusion of H₂S from the sediment into the water column. Finally, biofilm is observed at the top of the water column.

2.Column in the dark 

1. Control column    

STUDENTS:NENA KOUKOUGELI
                        DIMITRA-IOLI SKOUROLIAKOU
UNIVERSITY OF IOANNINA GREECE
BIOLOGICAL APLICATIONS AND TECHNOLOGY
AQUATIC MICROORGANISMS

WEEK: 6th

LEFT: control

RIGHT: warmth-exposed

The differences between the two columns are more obvious from the 5th to the 6th week.

In the first column, growth of urple sulfur bacteria in the middle zone is more odvious.

Purple sulfur bacteria are group of proteobacteria capable of photosynthesis and they are illuminated in anoxic zones. Unlike cyanobacteria or plants they do not use water as ther reducing agent, so they do not produce oxygen. Instead, they use hydrogen sulfide from the botom of the bottle which oxidised ti produse granule of elemental sulfure

In both columns, is still observed the presence of biofilms, the concentration of which, is greater in the first column

After 6 weeks, it has started a gradual distinction in the zones of the column, starting with the disply of sulphates purple bacteria. Considering that the forementioned zoning hasn't been yet observed in the 2nd column we can safely assume that it's groth rate is considerably slower. This happens probably due to the rapid fluctuation of temperature that affects the bacteria, not only directly by limiting the growth of the non-resistant, at high temperatures, bacteria, but also it affects them indirectly by the overheating of the plastic bottle that excretes damaging chamicals.

Hypothesis: The rapid fluctuation of the temperature on daily basis, reduces the growth rate of the microorganisms in the 2^nd Winogradsky column

Methane Production - Test with Fire

University of Ioannina

Team A (Gregoris Notarides & Kostas Karanicolas)

 

Tampakidou Maria-Ioanna Mpelonias Fivos University of Ioannina

Team 4 

Tampakidou Maria-Ioanna, 1479, Mpelonias Fivos, 1178 University of Ioannina 

5th-6th week observation:  11/11/2016 - 18/11/2016 

       The winograsky column-1 (contains eggs and it is closed) on the right has dark-black color that surely reveals anoxic conditions in the water. This is a sign of the presence of sulfate reducers and methanogens (archae). This column contains a lot of sulfide and very sandy sediment that allow the sulfide to diffuse easily into the bulk of the column, and as a result is anaerobic throughout. In addition, there are a lot of organics (great amount of eggs and paper) that make the decomposers increase. Many decomposers use sulfur, ferum and other kinds of anorganic matterials to oxidate organic matter and produce energy. In some of these reactions  there are byproducts such as hydrogen sulfide (H2S) and methane CH4 that cause the black color of the water. 

      On the other hand, in the winograsky column-2 (contains plasters and it is open) the water isn’t still clear anymore. It has been light green which is a sign of photosynthetic conditions. There is definatelly  some green algae and cyanobacteria , we can also see the biofilm which is products of microorganism witch after  the live there. Moreover, there are some organisms swimming in the water. They could be eterotrophs  and consume the autotrophs and their byproducts. Some of them have overall shape .The mud, is still totally black in some points and in some others is orange-red. It is a proof of red sulfur bacteria that finally are really observable. In the same time, the black spot are becoming bigger as sign of sulphur reducing bacteria such as Desulfovibrio that use products for anaerobic respiration using sulphates producing lots of H 2S. This H 2S will react with any iron producing black ferrous sulphide.

   

5th week