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Old 07-30-2019, 03:55 AM #1
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Winogradsky Column

Forewarning, this post has a little bit of discussion about what lead me to this post. If you care to skip it, the real content will be in the 2nd post.

So, I don’t know where the rabbit is that makes these holes all over the internet, but I ended up falling into another one today. After a good bit of reading and thinking, I did a search on here and RIU and didn’t see any mention of this topic. To tell the truth I’m very excited to bring this “new” topic to everyone’s attention here.

Like many of us here, I am a probiotic farmer. I try to get a healthy, diverse population living and thriving in my soil. I am successfully growing using only local materials, and making my own brews based on these materials. I have posted some of my use of these inputs in this thread (Local materials). I brewed random aerated teas for a while. Then I discovered LABs. I enjoy my experience growing them and they have given me a greater appreciation for knowing the microbes.

This lead me to seek out other microbes we can easily cultivate from the wild. If we look at a EM-1 recipe it contains
  • LABs
  • Yeasts
  • Photosynthetic Bacteria

The Yeast I have experience with from wine brewing and baking. I haven’t really figure out how having a bunch of yeast is a good thing yet, but I haven’t looked to much into it either. If anyone has some good information on this please share it.

The Photosynthetic Bacteria were completely new to me though. I knew they existed, but I didn’t recognize how beneficial they are to the soil. Further research lead me to Rhodopseudomonas palustris. This organism is AMAZING! It has 4 different modes of metabolism. Meaning it can live off a wide variety of materials, and in any environment. It even generates an electric charge. My mind was blown for a second there! After settling back down, I stated to look into how to get this thing into my soil. It’s remarkable adaptivity means we’re likely surrounded by it.

Onto the next step, how do we isolate and cultivate this organism? I have a background in biology, I have worked with agar numerous times for cultivating different species of bacteria and fungi. However, aside from taking a few swipes from my phone, or hand, or whatever I have always started with known inoculants. I know that would be the proper way to approach this. Unfortunately, I don’t know where to find those materials where I’m living, and I’m looking for something less involved. I am also not an expert in microbiology, especially identification.

Down this rabbit hole I came across Sergei Winogradsky. This is one cool dude, and I’m really surprised his name hasn’t been mentioned in all of this Probiotic/KNF buzz. He discovered lithographic bacteria way back in 1887. These guys chew up rocks and other minerals, and excrete them in more plant friendly forms. Of course we need some of those! He also discovered nitrogen fixing bacteria, photoautotrophs, and chemoautotrophs. If my boy Sergei could do it with what he had back then, surely we can do it today? And for real, why aren’t we giving this guy more props!
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Old 07-30-2019, 03:56 AM #2
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Winogradsky Column

Winogradsky worked his magic with a tool he invented called a Winogradsky column. This man was brilliant, he figured out how to make what I see as “column chromatography for useful bacteria”. This video does a great job explaining the concept.
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I AGREE


It seems logical, simple, cheap, and efficient. What more could you ask for? Hopefully it works as advertised!

I happen to have a small stream in my back yard, so off I went to make my own Winogradsky column.
  1. First you need to find somewhere water has saturated the ground for a while. This can be from the edge of a pond, lake, small stream, long standing mud puddles, and even brackish and salt water. This will be your inoculant source.


  2. Next dig up some mud. Everything I saw said to omit as much rock and organic matter as possible, you want mud! Unfortunately my river is pretty sandy, and the only thing I consider mud was in the middle. I waded in and gathered some mud. Just under the sediment there was a rich black layer. Every shovelful sent a load of bubbles rising to the surface. There was a slight stench of sulfur. We have life!


  3. The inputs required are minimal. You want to add a calcium source. I used powdered egg shells (1/2 tsp), but you could also use other sources or calcium carbonate, calcium sulfate or calmag. You also need a source of sulfur, I used 1/4 tsp magnesium sulfate (Epsom salt), but you could use calcium sulfate and I even saw an egg yolk was suggested. None of my research listed any proportions, and the YouTube examples just tossed in small amounts. I looked up how much sulphur is in a yolk, and a non scientific source said 25mg in the yolk and 50 in the white. Personally I don’t like the idea of using a raw egg, but I used it to kind of rough gauge of how much sulfur to add. You also need a carbon source. Many of the tutorials used shredded paper, I used shredded paper eggcrate. It’s something I had on hand, and I prefer to use this material since it’s already been recycled at least once. I imagine you could also use bio char, which is something I may experiment with if all goes well. Finally, you need a bit of water from your source, so make sure you collect some of that too. Mix all of the materials together EXCEPT THE WATER



  4. Fill up most of a clear container with mud. You can use plastic, but I think glass is preferable for me. A lot of things suggested 1L, but many examples used something smaller. My gut tells me bigger (taller) is better. I don’t currently have any 1L glass jars to devote to this, so I improvised with some pasta jars I had in the recycling. I made the effort to sterilize them by boiling hot water in the microwave. I was high, and it probably wasn’t necessary. Luckily I had let them cool before I collected any mud. The rest of the space I filled with water. I don’t know if it would be beneficial to leave a layer of air above the water. I could see how this could be beneficial as it’s another zone for microbes to grow. Again that’s something for further research.


  5. 5. Set your jar somewhere with sufficient indirect sunlight. Direct sunlight could cook your jar. Now like everything in this hobby, you just need to wait. The internet says you should see changes in 3-6 weeks. Changes that are visible, easily identifiable, and consistent. I sure hope everything on the internet is true!

Follow along with me over the next couple weeks, I’ll post how our jar is progressing. In the mean time, let’s discuss the different organisms I may or may not be cultivating. We should also consider how I may extract and use these organisms after this step. If you enjoyed this post, please click the “this post was helpful” button or leave me some rating. I know it doesn’t mean anything on the board or in real life, but I enjoy seeing that people are following along and appreciate my contributions to this website.

Thanks!

Last edited by Hookahhead; 07-30-2019 at 04:52 AM.. Reason: Should have previewed post
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Old 07-30-2019, 05:23 AM #3
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The video I posted above shows an idealized Winogradsky column. Looking at this image I realize an air space is a good idea. Also it’s not good to pressurize glass bottles, duh. I went out and removed the lids when I realized my mistake. However, my jars still lack the traditional airspace.



So let’s talk about the organisms found in the various layers.

Last edited by Hookahhead; 07-30-2019 at 06:06 AM..
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Old 07-30-2019, 05:45 AM #4
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I built one last year. Didn't let it age long enough. Take months to mature.

They do sell photosynthetic bacteria.

Sadly, the product I really wanted is impossible to get in Canada and it has 4 species in it.

It's called AZOO PSB SUPER RED PHOTOSYNTHETIC BACTERIA and it has these 4 bacteria in it.

Rhodobacter sphaerorides, Rhodobacter capsulatus, Rhodosprillium rubrum and Rhodospeudomonas palustris.

I just emailed somebody who may be able to send it to me. We'll see if I can finally get it.
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Old 07-30-2019, 06:50 AM #5
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Thanks for stopping by troutman. I’d be happy to hear any of your ideas, experiences or criticisms on this project.

The wiki has better info on construction than some of the other sources. I think I’m going to get a few more of these together in the next couple of days/weeks to test different inputs. It looks like we can swing the conditions to favor more specific groups/organisms. I like the idea of a small layer of sand over top of the mud, and a larger water area. I think as troutman pointed out the red/purple photosynthetic bacteria look like a good group to target.

Quote:
The column is a rough mixture of ingredients – exact measurements are not critical. A tall glass (30 cm long, >5 cm wide) is filled one third full of pond mud, omitting any sticks, debris, and air bubbles. Supplementation of ~0.25% w/w calcium carbonate and ~0.50% w/w calcium sulfate or sodium sulfate is required (ground eggshell and egg yolk respectively are rich in these minerals), mixed in with some shredded newspaper, filter paper or hay (for cellulose). An additional anaerobic layer, this time of unsupplemented mud, brings the container to two thirds full. Alternatively, some procedures call for sand to be used for the layer above the enriched sediment as to allow for easier observation and sampling of resulting populations. This is followed by water from the pond to saturate the mud (or sand) and occupy half the remaining volume. The column is sealed tightly to prevent evaporation of water and incubated for several months in strong natural light.

After the column is sealed tightly the anaerobic bacteria will develop first, including Clostridium spp. These anaerobic bacteria will consume the cellulose as an energy source. Once this commences they create CO2 that is used by other bacteria and thus the cycle begins. Eventually colour layers of different bacteria will appear in the column. At the bottom of the column will be black anaerobic H2S dominated zone with sulfur reducing bacteria, the layer above will be green sulfur photosynthetic anaerobic bacteria, then the layer will be purple which is sulphur anaerobic bacteria, followed by another column of purple anaerobic non-sulfur bacteria and at the top will be a layer of Cyanobacteria which is sulphur oxidising bacteria. This top layer of aerobic bacteria, produces CO2 which feeds back into the column creating further reaction.

While the Winogradsky column is an excellent tool to see whole communities of bacteria, it does not allow one to see the densities or individual bacterial colonies. It also takes a long time to complete its cycle. However its importance in environmental microbiology should not be overlooked and it is still an excellent tool to determine the major bacterial communities in a sample.[2]
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Old 07-30-2019, 08:11 AM #6
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Awesome Post!!!Very interesting! Thanks for sharing!!
Keep up the good work!!
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Old 07-30-2019, 12:31 PM #7
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Old 07-30-2019, 02:19 PM #8
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Hey Hooka; Great exercise for folks. You will find that I have recommended and referred to this process many times in the forum. Spurr did an exercise displayed with photos for growing PNSBs in similar fashion using fish [paste] as I recall.

Here is my most recent reference and a link to one of my heroes sites; https://www.icmag.com/ic/showpost.ph...3&postcount=30

Pitcher plants are another good source for PNSBs and fish scales.

EDIT; It appears that Jimmy Deacon's site is not available. He did say he no longer monitors it.
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Old 07-30-2019, 02:30 PM #9
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Hi MM thanks for stopping by, I was secretly hoping you would chime in! Wow, the post you linked is from the local materials thread I’ve been posting in, and somehow I still missed this info. I saw you reference PSB, but didn’t understand the acronym at the time. Also for some reason, a search for “Winogradsky” doesn’t turn up that post, maybe the basic search only covers thread titles? Anyhow, thanks for the info I’ll be sure to read up some more later today. If you have any insights, known issues, whatever please feel free to share them here.

Edit: It seems like the link you posted is down now. Here is a web archive of the link. https://web.archive.org/web/20190324...s/winograd.htm

If anyone else wants to set up a column and share their progress, this is the tread to do it!

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Old 07-30-2019, 02:36 PM #10
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EM1 has palustris in but not its relatives.

Edit: just noticed you basically mentioned that already

Great thread btw..
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