UCA_E7-8_1

WINOGRADSKY COLUMN (first day)

Rio San Pedro sediment + 0,3 g NaCl + 0,3 K3PO4 + 0,4 g sugar + 0,4 g CaSO4

The purpose of this practice is to observe the color change and the increase of microorganisms over time in a medium created by ourselves with the reactives elements mentioned above.

We introduce the same mixture of the same quantity into two different tubes. One of them will be exposed to light and the other will not.

Then, we will explain why we have added each of the aforementioned compounds:

• The sucrose is added to bring carbon to the medium as every living thing needs a source from which it can obtain it.
• CaSO4 provides sulfur to the medium. It will allow energy-capturing bacteria to grow through the reduction of this.
• With the addition of K3PO4 we are contributing with phosphorus to the medium, which is used by microorganisms for their metabolism and it is an essential macronutrient for the growth and development of living beings.
• We added NaCl to have a more saline medium than we had originally, and thus favor the growth of halophilic bacteria.

          

In this image we can observe the state of our tubes immediately after the deposition of the mixture in them and therefore they present a homogeneous and compact color throughout the column.

The tube that is in contact with the sun has a different development than the one that is not.

Over time, we will see different colors and changes in our column due to the growth of microorganisms. These variations can be seen because each bacteria will grow under the conditions that are most favorable to them along the column.

Ana García Ramos, Carlota Borne Bernal, Ernesto Segundo Mendoza, Maria del Carmen Espinosa Corona and Pablo Carrasco Ercilla

Second post: The evolution of our Winogradsky column. Group A4.

In this post we will discuss the progress of our column during the last four weeks.

First week:
Four parts are distinguished:
At the bottom of the tube we can see bubbles which expands to the Surface . It seems like a volcano.
Due to the presence of an acumulation of organic material, the microorganisms make methanogenesis, althought this is one of the less efficient source of energy. This process produce the grey colour .
Above the grey colour, we can observe a black area where oxigen can´t access ,so the microorganisms who live there are anaerobic and instead of using this molecule they take the sulfate and release  sulfuric acid and carbon dioxide. The sulfuric acid reacts with iron producing iron sulfate, which is a black precipitate.
In the third zone, the orange and black colours are mixed . This part can be reached by oxigen , so microorganisms are aerobic and organoheterothophs. The orange colour is a consequence of the photosinthetic pigments.
In the last part, at the top of the column, there is some orange water without any algae but in the surface there is an oily liquid due to the bacteria.

                      
Second week:
This time, we can distinguish three parts:
The bottom of the tube has stayed grey, due to the methanogenesis, as the last week.
The orange liquid of the surface has not change neither.
The principal changes have occured in the black zone of the tube (at the middle). It has expanded, because of the acumulation of organic material.
At the surface of the  mud, a green colour has appeared which indicates presence of photosinthetic microalgae.
                                         
Third week:
The black zone of the bottom of the tube has not changed. However, in the middle of the tube a light pink colour has appeared, due to the presence of a special photosintetic bacteria. It assimilates sulphur and produce sulfate instead of oxigen.
At the surface, the orange colour is mixed with green and brown. As we said, the green colour has appeared because of the growth of photosinthetic microalgae.

                   

Fourth week:
The major change is that the pink colour has become more intense and dark, because the bacteria has increased the concentration of sulfate.
At the same time, it has appeared bubbles at the surface because of the presence of oxigen produced by photosinthetic microalgae and cianobacteria.
The gas produced in the methanogenesis has created some breaks which we eliminated by hitting the table with the column.
The column was warm due to the biological activity and the solar heat.
In general, we have noticed a disgusting smell every week because of the excess of sulphur.

                    

Winogradsky Column Day 0. Group A5

We added into the column:

- Pond mud: 24.7 g.
- Paper (5 pieces): 0.5 g aprox.
- Yeast: 0.4 g.
- Calcium carbonate: 0.3 g.
- Sea water.

The mud will be the medium of the column where the microorganism will grow. Each component will apport some nutrient for each kind of microorganism.

The paper will be used as the source of glucose by the chemotrophic microorganism but, at the same time, it will only permit to grow the microorganism that can digest the cellulose. The yeast offers a lot of nutrients due to its amount of amino acids. Litotrophic microorganism will grow using the calcium carbonate as the carbon source. It is a saline medium because of the sea water so only will grow microorganism that resist those conditions.

At the end, there will appear different layers. Each colour that appears in the column will mean the growth of different microorganism. At the bottom of the column, will grow anaerobic or facultative anaerobic microorganism. Also, the sun light will be used by autotrophic microorgnanism that synthesize glucose by photosynthesis (cianobacteria).

WINOGRADSKY COLUMN DAY 0: HYPOTHESIS (GROUP 6C)

Pond mud→ 20,81g

Yeast extract→ 0,1g

Sodium sulfate→ 0,4g

Calcium sulfate → 0,1g

Pieces of paper→ 0,1g

For our Winogradsky column we use pond mud as a culture medium. The yeast extract can  be used for microorganisms as a nutrient source because  it is high in  B vitamin, and it contains amino acids and other factors  that are ideal for their growth.In addition,  we have used sodium sulfate and calcium sulfate as a source of sulfur that will be used for the anaerobic microorganisms that grow at the bottom of our column in order to being able to accomplish their breathing process.Finally,we have added little pieces of paper that can be  used as a carbon source (cellulose) so the organisms can obtain  energy from the synthesis of glucose which is  required for their biological processes.

As a result of all this, we predict that there will be able to notice  a few layers.We will be able to find aerobic microorganisms as there is enough oxygen for them to use. Facultative anaerobic organisms will be found in the layers that are in between, and anaerobic organisms will be found in the deepest layer of our column.

Winoblog, Third post, group B7.

          This is the third post which we make about our “Winogradsky column”.

       By this time, the column has changed a lot. For example, microalgae have appeared in our column. To be more accurate, they have appeared in a water’s zone with oxygen.

        

           On the one hand, some gas bubbles have appeared in our column. That bubbles 
as a sign of metabolic activitiy. Cianobacteria produces oxygen using another componets of our “widnogradsky column”.

        On the other hand, stratification can be better appreciated. For example, under the water, there are a large area of ​​greenish gray. This is because microorganisms are growing up there, more specifically Beggiatoa is growing there.

         To sum up, we can se how the bio-system that we have created is becoming stable. Besides, we can appreciate differents strata that a couple of days before there werent there. 

Figure 1: Microalgae.

Figure 2: Photo taken 17th April.

UCA_3A_2: Day 48 - Our column in process.

In the first week, we can see a black colour at the bottom of the column. This is a result of sulphydric production. Besides, there is water in the top of the column, where we can find an orangey brown colour (this is the zone provided with oxygen).  Moreover, consequently to have added a lot of mud and just a bit of sand, there is a part in the middle of the column that is completely grey and homogenous, where gases can’t pass through it. Due to this point, we can guess that there will be two separated communities.

In the second week, we observe that the homogenous part has decreased. Besides, we see a black colour in the top of the mud (there are bacteria that feed of sulphate and produce sulphydric acid). In another hand, the brown community are bacteria that are able to use oxygen.

After two weeks, we focus on our column again. As a result of have added a lot of mud, the development of the microorganisms  is very slow. The continuous black colour indicate us that bacteria keep producing H₂S. Now, there is an orange colour that is a signal of the existence of photoheterotrophs microorganisms producers of iron.  

Winoblog, Second post, group B7.

            In this second post, we can appreciate a change in the colour of our “Winogradsky column”. The methane wall cavity has been deleted, because organic substance’s excess was consumed. Besides, is possible to see muddy water. 

            In addition, a rosy colour appears, because a microorganism like Chromatium uses CO₂ and H₂S to create organic molecules. On top of rosy stratum, we can see other stratum more light where we find Rhodomicrobium. Rhodomicrobium uses CO₂ and organic acids to produce organic molecules.

             Finally, at the beginning of Winogradsky column, there is a space that has oxygen. This oxygen is utilized for filamentous microorganism like Beggiatoa, it produces sulfuric acid and organic molecules. Besides that, on wáter, photosynthetic mirobes make their work.

Figure 1: A rosy stratum.

Figure 2: Photo taken 31st March.

Figure 3: Space with oxygen.

UCA_C2_1: Day 1 - Our Winogradsky column

Hi! We are Irene, Raúl and Nieves. We are studying Biotechnology in Cádiz and we want to show and explain you how we prepares our Winogradsky column. On later posts, we will see some changes due to the growth of many microorganisms.
Our column is made of:

• Rio San Pedro sediment (mud and sand)
• 0,23 g sugar
• 0,02 g filter paper
• 0,40 g CaSO4

We have decided to add sugar and filter paper in order to give microorganisms a carbon source to do fermentation. They will release some waste products that will be used by another microorganisms to do anaerobic breathing, so they could use CaSO4 (sulphate source) to break down into H2S. This sulphide will be used too by another ones, and so on.
We think that our column is a gradient of O2 and H2S, so we will appreciate different types of microorganisms (aerobic and anaerobic ones) distinguished by colours. At the top of the column, where there is a lot of O2, aerobic microorganisms will grow. Whenever we descend to the bottom, the amount of H2S increases and O2 disappear, so it favours anaerobic microorganisms growth.
We haven't taken photos of the column the first day, but we will see the evolution on the next posts.

UCA_4A_Entrada 1: 1º day

Hi there! We are Elena, Pepi and Marta and we are going to explain how we did our column in the lab. First of all, we mixed  20,9 g of mud with some seawater until we had an homogenean mixture. Then, we added :
- 0,41 g of CaSO₄ because it is a source of sulfur for  aminoacids. SO₄^2-  + O.M  à  CO₂ + H₂S^-

- 0,21 g of NaCl because we want halophilic bacteria to grow.
- 0,58 g of CaCO₃ so autotrophic bacteria can grow.
- 0,31 g of C₁₂H₂₂O₁₁, which is a common carbon natural source and also allows heterotrophic microorganisms to grow.
- 0,22 g of cellulose, another carbon natural source that also allows heterotrophic microorganisms to grow.
- 0,1 g of agar, a polysaccharide  that attatches cells.
Now we introduced the mixture in our test tube.
After that, we measured 30 g of mud and we mixed it with 90 g of sand, and we added this new mixture to the test tube too.
We also added a bit of seawater at the top of the test tube and left it in the lab for one week.
The results we expect to get is a concentration gradient that shows the different types of microorganisms that grows in our Winogradsky Column.

Winogradski Column. First post

Hello we are David , Diego and Malena, we are Biotechnology students. We have prepared a Winogradsky column and we are going to show the changes that have been produced. 

The Winogradsky column is a microorganism cultivating resource. The base components we use are :

- Sand 75 gr.

-Mud 25 gr.

-Thin layer of sea water 

This components are the same for every columns. Moreover we decided to add :

-0.13 gr of sugar as a carbon source

-0.10 gr of calcium sulfate as a sulfur source

- 0.09 gr of  yeast to produce fermentation.At the bottom of the column there is no oxygen, so it will be a different  source of organic matter,for example , lactic acid.

We will try to make a gradient of O2 at the top and H2S at the bottom so that both aerobic and anaerobic organism can grow.

Our hypothesis is that heterotroph organism will grow because of the external carbon addition and also sulfur fixative bacteria because of the sulfur source.