Winogradsky column

Winogradsky Column

Our group is formed by a group of students of biotechnology (Marta,Chiara and Juan) and we are so pleased to show you the development of our Winogradsky column along several weeks.

Raw materials.
-NaCl, as a source of salt (0.4g)
-Mug, like the habitat of the bacteria (20g)
-CaCO3,as a source of calcium, (0.46g)
-Na2SO4, it releases energy in water (0.42g)
-Paper, as a source of carbon (0.3g)
 -Yeast, it allows bacteria to do fermentation (0.41g)

Hypothesis:
- The paper is one of the most important elements in the increasing of the heterotrophic microorganism because it´s their one source of organic carbon. This kind of organisms can´t produce photosynthesis.
- The ion SO4- can be the source of aminoacids like cysteine or methionine. There are a kind of  chemoautotrophs organisms that use the energy of the oxidation reaction (the oxidation of the sulfur). They don´t need the oxygen to live.
- The yeast creates a culture medium rich in energy thanks to the process of fermentation.
- The mug is rich in ammonium salts, that are one of the main sources of nitrogen, a very important compound of aminoacids and nucleic acids. There are a kind of bacterias that fix the nitrogen of the soil, for example the Rhizobium.
We think that is very posible the growing of heterotrophic, Sulphurous, Nitrifyng, fermenters bacterias.


From the begining the whole column were black and we could not notice any difference in the organisation


First week.

In the first week we can see a different organisation, we can see tree parts;The brown one on the top of the column(with some air caused by fermentation),a black one in the middle and a grey one in the bottom.

Second week.

We notice that the black part has increased making a gap of air because of fermentation.The organisation remains equal and there is far more air in the whole column.

Third week

 

 In the third week the structure is the same as in the previous week, but we notice that some seaweeds have grown up on the top of the column,where is oxygen that is needed by it,because of CaCO3,and it is important to underline that in the middle of the column has appear SO4(2-) precipitated.

UNIVERSITY OF IOANNINA GREECE
BIOLOGICAL APLICATIONS AND TECHNOLOGY

COURSE: AQUATIC MICROORGANISMS

Team 9: Katerina Mironaki
             Levidiotis Charalampos

Hello everyone!

       Inevitably our semester long project comes to an end. We tried to delay our entry in hopes of having clearer and more definitive results. So without further ado, these are some pictures of our columns about 17 weeks from the beginning of the experiment. 

       Thankfully the results followed our prior assumptions “a discernable growth spurt of microbes in the second column much sooner than the first one as well a more intense coloration of the column. “        
       By comparing the two columns someone can easily observe the advanced growth state that characterizes the second one. From the intense coloration of the water to the early appearance of the colonies of purple sulfur bacteria the second column kept surprising us. Day after day, week after week new things started to make their appearance (new colonies of microbes formed speckles and strips, the sediment began enriching, the bottle swelling etc). Some of these new “findings” cowardly appeared in the first column but with a notable delay in time.

       In the end we can say that we are pleased with our results but we needed more time(a pity that semesters aren't longer...not really). Nevertheless we learned a lot of things and had quite some fun during these months.

       

 Everyone, thanks and good luck with your own projects!   

                                                          Column 1

                  

                           

                                                              Column 2

Tzani Kalliopi

Mavridi Olga

University of Ioannina Greece
Biological Application & Technology

WEEK: 8th

Left: Winogradsky column with egg

Middle: Winogradsky column with garlic

Right: Winogradsky column with egg & garlic 

The hypothesis stated was confirmed; we observed differentiation between the three bottles in microorganisms’ development. Specifically, in the bottle with the egg as a sulfur source, we saw green sulfur bacteria characterized by a green/olive colored zone, and red-purple sulfur bacteria growing. We also saw the formation of biofilm. In the bottle with garlic as a sulfur source, we saw green sulfur bacteria and red-purple sulfur. However, we also observed the formation of FeS. Also, the water was easier to observe, since there was little to none biofilm, and we saw the growth of a plant. Finally, in the third bottle, which had both egg and garlic, we saw green sulfur bacteria, red-purple sulfur bacteria, and purple non-sulfur bacteria characterized by a red/ orange or rust colored zone. Again, in the water, there was no biofilm formation, and it had a high cyanobacterial growth.

Hello fellow winobloggers! We are Kalliopi Tzani & Olga Mavridi, under graduate students in the field of Biological Applications & Technology in the University of Ioannina. In terms of a class/ curriculum concerning aquatic microorganisms, we were assigned to make a Winogradsky column. For this purpose, we placed lake soil and lake water, from the Vrelis’s lake, into three bottles in a ratio 2:1. Each bottle contained lake soil and newspaper as source of cellulose. The bottles, however, contained different sources of Sulfur. In the first one, we added an egg, with its pod. The second one contained garlic & the third one contained both of these sources of sulfur (egg & garlic). Our bottles were closed, as to not exchange gasses like oxygen, with the environment, and they were placed in a sunny room. We observed them in a weekly base and we stated the following hypothesis: if there will be a differentiation between the 3 bottles concerning microorganisms’ development.

Vrelis's pond

Winogradsky column with egg & garlic (t=0)

Winogradsky column with garlic (t=0)

Winogradsky column with egg (t=0)

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)