Fred Transplant: Success!

A Gram-stained view of yeasts and bacteria in a sourdough culture named 'Fred'.

I had to do a Fred Transplant last week. A grey fuzzy mold had taken up residence in on the sides of the jar above Fred’s liquid culture, so I set up a fresh container with fresh water and flour, and dipped a spoon down the center of Fred to the bottom, pulling up just a tiny amount of the stuff in there. Then I mixed it into the fresh stuff and covered it with plastic wrap (instead of a paper towel this time.)

Fred smells like Swiss Cheese Feet right now, but he’s obviously still growing, as you can see from last night’s “Gram Stain” microscopy. The slightly blurry light-red-brown lumps are, I believe, yeast cells, possibly Saccharomyces boulardii, since I dumped a capsule of supposedly-still-viable “probiotic” yeast of that species into Fred previously. I have no idea who the bacteria are in here at the moment. I did also see a small number of longer, thinner bacterial cells in there (presumably Lactobacillus) though most of them are the ones you see here.

Meanwhile, I’m about to dig out the still-unused Hillbilly Autoclave and try it out on the media I’m mixing up to try to obtain a culture of genuine wild “native flora” vinegar/kombucha yeast-and-bacteria to play with from the local wildflowers that are just now getting into full bloom.

My starting recipe goes something like this: I mix up about 2 Liters of distilled water with about 100g of glucose (“Dextrose”/”Corn Sugar”), 100g of sucrose, 500mg of L-Arginine, and enough phosphoric acid to drop the pH down to about 5.5 to 6.0. That is intended to be then poured into small “canning” jars in about 100ml amounts and pressure-cooked for at least 15 minutes to sufficiently sterilize and seal them. Meanwhile, a single generic-brand children’s chewable vitamin is crushed up and dumped into a 4-oz bottle of cheap vodka and well shaken.

Then when it comes time to go bioprospecting, I’ll pop open the jar of acidic sugar solution and add about 5ml of the cheap-vitamin-vodka to it to give me about 2% ethanol, and then go find some flowers to cut off and dump into the jars, which will be loosely covered with foil (to let air in but keep dust out) and put in a nice quiet cupboard to grow for a few days.

Hypothetically, the only things that are likely to grow in that will be microorganisms associated with vinegar-making. At some point I’ll also make up a batch of sweet black tea and see if I get a kombucha-like culture going in it, and make up some solid media to try to isolate individual microbes from it.


Looks like the truckloads of candy-seeking larvae are done finally. Wretched little urchins now get driven from block to block rather than walking the neighborhood like we did.

(It doesn’t actually bother me as much as that makes it sound, I just like having an excuse to say “wretched little urchins”…which reminds me – I have only about a month to get a cheerful flashing “Bah! Humbug!” sign built…)

The only thing that really annoyed me is the fact that having to be ready to be interrupted by another horde of costumed consumers meant I couldn’t really spend any of the evening getting into anything requiring any real attention…which means the 113g of CaCl2.2H2O I’ve got sitting here now to go with my Xanthan Gum has been left neglected, and I still don’t know if I can make Xanthan Gum gel into beads the way you can with sodium alginate. I figure it must be possible, given that both Xanthan gum and Alginate (among others) were all formed into little “bio-booger” beads using the same kind of process in the paper I discussed in Episode 2 of my little audio oggcast. Perhaps I’ll have time to find out tomorrow.

For now, it’s time for bed. Daylight Losings Time starts tonight, so if the critters allow me to actually sleep, I ought to be well rested to attempt some serious lake-spanking in the morning – there’s supposedly a resort on the shore of the lake that has a sushi bar, and the idea of being able to paddle out for sushi amuses me. It looks like it’s at least 9-10 miles away, though, so it’ll be a long trip if I attempt it. Hopefully I’ll have time left after that.

Also, the developer of the libdmtx datamatrix barcode encoder and decoder software posted a recent comment on my previous post about the software and its potential uses – looks like some interesting projects going on there, including one intended to generate ID cards that only legitimate authorities could read (so as to prevent identity theft).

P.S. Anybody know how to build a really good (but simple) ozone generator for sanitization purposes? Or the effective pore sizes of commonly available materials like plastic wrap? Or if a corporate entity can be a shareholder/partner in a Limited Liability Company?

Don’t forget to feed and walk your mitochondria

Yes, I’m still here – though I don’t know if any of YOU are.

The pay at my job is somewhat low for the skillset it requires, but makes up for that by having a very reasonable workload, a pleasant work environment, and certain perks – like access to the electronic journals that my employer subscribes to. I added an RSS feed from pubmed intended to cover my main interests – basically edible and industrial microbiology and biotechnology. Every day, a list of 300-600 or so new scientific articles pops up in my feedreader and I scan through the titles looking for anything interesting to me. Unintentionally, my selection appears to also result in quite a bit of diabetes, obesity, and sports medicine research. Lately I’ve taken a moderate interest in our own most blatantly bacterial components, the mitochondria.

Mitochondria are kind of like a nearly 2-billion-year-long case of typhus (or Rocky Mountain Spotted Fever, if you prefer). After infecting our ancestors (and now us) for so long, they’ve been reduced to dependency on living in our cells. Perhaps a bit like the progression from wolves to Chinese Crested dogs. On the other hand, having thoroughly domesticated them, we get a lot of use out of them, and couldn’t live without them. Their ability to harness the electron-sucking power of oxygen means we get almost 20 times more energy out of our food than we otherwise would, which is a good thing since biologically speaking, keeping the hideously complicated mess of biochemistry that makes up a human body takes a ridiculous amount of biochemical energy compared to that of normal organisms (i.e. prokaryotes).

Lately in the stream of new publications I’ve been seeing a number of papers suggesting that a lack of proper mitochondrial activity might be related to obesity and related problems (e.g. “metabolic syndrome”, type 2 diabetes and insulin resistance, obesity-related “inflammation”, and so on) and even some age-related problems, both physical and mental. There is some seriously interesting research going on into treatments to potentially stimulate mitochondrial activity and whether this might help solve a number of health problems.

So…take good care of your mitochondria. For the past couple of weeks I’ve been trying to pay special attention to properly feeding my mitochondria and making sure I take them for regular walks (and paddling trips and so on). It could, of course, be purely psychosomatic, but right now I feel better than James Brown

There’s a fair amount of rational skepticism over using drugs or nutritional supplements to stimulate mitochondria, but here’s a tip that I suspect everyone’s doctor would accept: make sure you take your mitochondria for regular walks. Frequent exercise (particularly endurance exercise) seems to be a scientifically well-accepted way to induce production of more mitochondria.

But now I have to go to bed. My main complaint with work these days is that it eats up essentially my entire day, leaving me with just enough time for some household chores between getting up in the morning and going to bed in the evening. Not their fault I live almost and hour and a half from work, though (and at least the commute is through relatively low-traffic and scenic terrain.). Still, it makes it hard to get blog posts and podcasts done (episode 4, on the subject of “heat-fixing” of bacteria for microscopy – particularly Mycobacterium tuberculosis – will be out as soon as I can manage. Still pondering the subject of Episode 5. I’m saving the “Two Mass Spectrometers, High Performance Liquid Chromatography, and a Female Donkey” episode for later when I manage to surpass the “nearly 3” listeners that I seem to be stuck at…)

“Improvements in the Fermentation and Maturation of Beers”

Judging by my webserver’s logs, almost nobody actually bothers to click through the blog-carnival host’s site to read my Giant’s Shoulders” posts. This could be due to a secret conspiracy involving famous bloggers and several shadowy government agencies. I suppose, though, that there’s a chance that simply nobody but me is that interested in non-medical microbiology. Well…today’s post is an attempt to disprove that concept, for what aspect of non-medical microbiology could be more universally appealing than beer?

Unfortunately, in the middle of trying to assemble this posting, I see the February host has decided to put the carnival up a day early, undercutting my experiment. See, I told you it was a conspiracy! I suspect the Secret Cabal of Popular Bloggers was getting pressure from the Trilateral Comission, the NSA, and Pepsico® to silence me, so they had to do it. At least being forced to miss one, I am now free from the “I’ve been posting to these since the beginning, I can’t miss one now!” treadmill.

That means, loyal readers, that you get to see this post a month before everyone else! Hooray! Stick it to The Man™! Comically paranoid rantings aside, it also means I can split this up into more than one post, which may be more readable considering how much ground the article in question actually covers. Today’s Classic Scientific Paper is:

Nathan, L:”Improvements in the fermentation and maturation of beers.”; 1930; J. Inst. Brewing; 36; pp538-550

I ran across this reference recently while working my way through an industrial microbiology text[1] that I checked out of the campus library. According to the author of this text, “The use of cylindro-conical vessels in the brewing of lager was first proposed by Nathan (1930)[…]”, referring to the now-ubiquitous style of metal fermenter seen in small brewpubs and “MegaBladderwashCo” large-scale industrial breweries alike. Based on this I had expected the reference to be a digression on the design, construction, and testing of the fermenter. When inter-library loan managed to get me a copy of the paper, I found something much more involved.

The paper is a presentation made by Dr. Leopold Nathan in 1930 to the Scottish section of the Institute of Brewing. The topic was not simply a fermenter design but the entire “Nathan System” of brewing which appears to be the basis of modern large-scale brewing, especially for Lager-type beers. At this point, Dr. Nathan had apparently already been developing this system for about thirty years (apparently starting with a German patent in 1908, which I’ve yet to find a copy of), so as you might guess it was not just a single invention but a whole collection of them. Compared to the more rustic techniques frequently in use at the time, the “Nathan System” of brewing promised to provide faster production, more consistent results, and a better final product. It does this mainly by improving the removal of “trub” (the cloudy bits of protein and such that settle out of the malt-water – the “wort” – after you boil it), preventing infection of the beer with undesirable organisms during the cooling, hops-infusion, and aeration, and by eliminating the need to “age” the brew to make it palatable. The most important improvement in the “Nathan Process” seems to be how he treats the wort between boiling and “pitching”.

For anyone unfamiliar with the brewing process, here’s a Grossly Oversimplified review of the steps:

  • Boil some malt-sugar dissolved in water to sterilize it and to help coagulate the “trub” proteins so they’ll settle out of the liquid.
  • Cool the malt solution and aerate it so that the yeast will grow in it.
  • “Pitch” your yeast into the now-cooled-and-aerated malt-water, in a container that will keep air out while letting out the carbon dioxide bubbles that the yeast will give of during the fermentation
  • Wait until the yeast get done fermenting, then put the resulting liquid into bottles/kegs/casks/whatever.

Diagram showing the containment vessel, cooling system, and sterile-air generator for the 'Nathan method' of brewing
I’ve added a couple of labels to that image from the paper, which I’m guessing was itself copied from a contemporary patent of Dr. Nathan’s. There are two purposes to this part of the Nathan Process – To cool and aerate the wort quickly without exposing it to risk of contamination, and to move trub and volatile sulfur compounds that would otherwise make the brew taste and smell funny. The hot boiled wort is pumped directly into an insulated vat (labelled “A” in the diagram) from the boiling kettle. At this stage the wort is hot enough to prevent anything from landing in it and growing. Then, the hot wort is pumped from the top of this vat into a clean-room containing a cooling device that the wort is poured on, cooling and aerating it as it flows through. Infection is prevented here by the fact that the room has a continuous stream of “sterilized” (or at least well-filtered) air, which is exhausted through the vent in the ceiling. The cooled, aerated wort is then pumped back out of the room and into the bottom of the insulated container below the still-hot wort.

Because of the large open cooling room with its constant stream of clean air, the cooling and aeration step also allows the volatile sulfurous compounds of “jungbukett” (The “Bouquet of Youth”; the unpleasant smells and tastes of immature beer, described in this paper as ‘onion-like’) to evaporate off and be carried away. Since waiting for these compounds to break down was apparently a primary reason for having to “age” lager before selling it, this not only improves the quality but eliminates the need to store the beer for months after fermentation.

The now-chilled wort then rests back in vat “A” and the trub settles out onto horizontal plates inside the vat, where it stays behind when the clarified wort is pumped out to the fermenters.

I did some poking around, and this appears to be what is described in US Patent# 1,581,194 (application filed in August of 1921), in case you are bored and want to look that up. If not, or if you don’t want to deal with the frustrating hassle of trying to view TIFF files in your browser, I intend to provide a followup post with some more details of the process and some interesting bits I found in it, and I’ll include a pdf of the patents, assuming anyone wants them.

Oh, one last thing – I’ve had no luck getting any biographical information about Dr. Leopold Nathan. Unfortunately when you search for “Leopold Nathan”, the results are clogged with references to a murdering smartass named “Nathan Leopold” instead. Doesn’t Google™ realize that brewmeisters are far more important than obscure murderers? No pictures of him, either, so I can’t even say whether his hairstyle is cooler than Eduard Buchner’s or not.

[1] Stanbury PF, Whitaker A, Hall SJ:”Principles of Fermentation Technology (2nd edition)”; 1995; Elsevier Science, Ltd; Tarrytown NY


They say “When life gives you lemons, make lemonade”. What if life gives you snot instead?

Someone's slime-covered handNow, see, this is what happens when I’m too poor to buy nice distracting new toys for myself. (No, not the hand in the picture – that’s not mine, it’s just there for illustration.)

I found a pot that I’d rinsed well but then left filled with water in the sink to soak, to help remove the last of the rice bits stuck to it. It hadn’t gotten stinky or fuzzy or anything, but it had gone…viscous. Like a light sewing-machine oil. Naturally, I took appropriate action to deal with it.

I fed it.

Glucose (“dextrose”), to be precise. It’s since been dumped into an old glass jar and the original pot thoroughly scrubbed with hot soapy water. At this point (a day later) the slime is closer to the viscosity of vegetable oil now. And I fed it again.

I wonder what it is? I mean, obviously it’s bacteria-snot, but what kind? I suppose if I had some iodine I could check to see if it’s a polysaccharide (evidently this test works on polysaccharides besides starch). If only I had a microscope, I could at least get some basic hints as to what’s producing the slime. Maybe I can maintain a culture and figure it out later, if I can ever afford a real microscope. Perhaps I could even attempt a strain-improvement program to increase the production rate…

Uh…I did mention I was a nerd, right? Okay then.

I wonder if anyone at work has a bacteriological microscope setup that I could use?…

I have a shocking confession to make.

I think I may be a nerd.

No, no, don’t try to deny it. I think it pretty much has to be true for someone who reads a 10-page scientific paper (hover or click here to see the reference) in order to learn that bacteria-snot is slimy. Yes, I am a nerd. And for that I am deeply, deeply….

Oh, who am I kidding? I like being a nerd.

Stainless Steel fermenters at a breweryFor one thing, being a nerd allows me to fully enjoy one of the perks that my job gives me – namely access to a lot of scientific papers that I otherwise wouldn’t be able to afford to obtain access to from the greedy [insert your favority profanity here] who insist on charging $30 for permission to look at a decades-old articles for a day. I should add that this perk includes Inter-Library Loan for articles that I can’t get online, and the service on campus is great so far. Same day delivery of a classic article from 1930 in what I’m guessing most people would probably consider an obscure journal.

It doesn’t have quite the same thing in it that I expected from the source that pointed me to it, but I think it can still be considered “classic”. I need to re-read it more carefully to make a final decision on this, but I think I have my next “The Giant’s Shoulders” article in time for this month’s upcoming issue. And, yes, the picture attached to this post is a hint (and, no, it’s not directly related to the bacteria-snot article mentioned above in any way…)

Coming Soon to a blog post near you!

I got a little money for Christmas, but I’m feeling quite guilty as I didn’t really have the time and money to reciprocate in advance. I also haven’t been able to figure out what to spend it on until now. Now, I have a solution for both problems.

Xanthomonas campestris growing in a petri dish and exuding slimeI intend to spend it all on fermentation-related food ingredients and do some experimentation with sweet-tasting foods. Initially, in addition to flavors (spices and whatnot), I need to track down bulk quantities of:

  • Erythritol, which is a virtually non-caloric sugar alcohol which unlike sorbitol and so forth is not normally prone to cause gastric distress, and unlike Xylitol is not hazardous for beloved household pets. Better still, it actually is very tasty unlike that nasty hippy “Stevia” crap (which isn’t produced by fermentation anyway, as far as I know).
  • Food-grade Glycerol (“Glycerine”), which I hypothesize is close enough to the structure of Erythritol to mix well with it and help the erythritol dissolve (and hopefully prevent crystallization, much the same way the “corn syrup” does with sucrose).
  • Xanthan Gum. MMmmmmm…edible bacteria-snot. (Okay, for all that this sounds disgusting, it’s really somewhat similar to pectin, which like xanthan gum is a polysaccharide. Pectin is just fruit-snot rather than bacteria snot. Dietarily, both count as “soluble fiber”.)

There are probably other ingredients I can come up with as well. For those of you out there who are owed gifts: Chewy candies, hard candies, baked goods, and/or beverages, what’s your preference for my initial experimentation? Assuming anyone’s interested, I will probably blog my results…

Meanwhile, I’ve also been thinking about geolocation, geotagging of audio and video media, and Asterisk again. I want to take the lessons learned from my playing with the “Where Was I?” prototype and turn it into a real geolocation system, integrating with Asterisk and Laconica (which turned out to be easier to set up than I’d feared – I’ve now got my own Laconica server at, though I need to sit down and activate the IM integration (Twitter may have abandoned IM, but it reportedly works fine in Laconica).

It also turns out that you can use Asterisk with cellphones(!), at least if they have bluetooth. That’s handy to know…

Argh – too much to do, not enough time!…

Obscure scientific papers, Mad Science, Travel, and other randomness

First – an amazingly astute observation that I’m ashamed to have not previously noticed myself (click image to go to it’s original site and see it full-size…):
Most 'Mad Scientists' are actually just 'mad engineers'...

I’m proud to say that I think testing Mad Hypotheses is great, and will continue to try to be a Mad Scientist. And a “Dirty Old Man” someday, but that’s a whole separate issue.

Second – I am really loving the perks of my new job – namely access to the college library system. I had previously mentioned (see last couple of paragraphs) a certain article that I wanted to get my hands on:

Greenberg LA:”The Definition of an Intoxicating Beverage”;Q J Stud Alcohol. 1955 Jun;16(2):316-25

Not only does the medical library have copies of a Czechoslovakian microbiology journal, the main library had a set of this old journal, too. I have my bedtime reading for tonight…

Thirdly – Another Giant’s Shoulders carnival has come and gone. I now believe that Eduard Buchner had hit upon not only a useful truth of living systems, but also a nifty alternative “mad scientist” hairstyle. Now I need to come up with one for next month. It’s been getting me thinking, though. That blog carnival is intended for “Classic” papers. Implied is that the papers are somehow important to the development of some scientific field or other. I’d like to see a variation on the “old papers” theme focussing on random old papers (where “old” might mean a few years or decades, depending on the subject) that people have found useful or interesting. Stuff that isn’t necessarily ground-breaking and has perhaps been forgotten or lost to obscurity but still has useful things to teach us. Naturally, I’m thinking especially Microbiology (and especially Microbiology other than Medicine) and Food Science. The Carnival could be called something like “Second Chance Science” or something of the sort. Just a thought.

Fourth – speaking of “Microbiology Other Than Medicine” and Food Science, apparently The National Academies of Science want to know what scientific topics people most want to read about. As usual, “microbiology” appears to have been relegated in their breakdown to merely a subset of either medicine/diseases, “biology”, and perhaps a small subset of “energy” and “Feeding the World” (no, seriously). The survey includes space to tell them what they’re missing – I heartily encourage anyone who cares to make sure you take the survey, and mention industrial and environmental microbiology and food science as subjects they shouldn’t continue to neglect.

And, finally – next week I need to make a very-long-overdue run back up to Idaho to grab some things from the old house and make sure it’s still standing, the water’s really turned off, nothing unnecessary is running, etc. 1600 miles of driving each way. Ugh. Anybody got any good recommendations for things to listen to on the trip? Other than having a chance to finally grab some things that I am missing, maybe I’ll at least have a chance to visit New Belgium Brewing Company again, since my route goes right past it. So long as I’m not driving by on Christmas day (when I assume they’ll be closed) I may have a chance.

Über alkoholische Gärung ohne Hefezellen

In my last submission to “The Giants’ Shoulders” blog carnival, we saw how the famous surgeon Dr. Joseph Lister deftly demonstrated definitively that fermentation processes were caused by live microbes rather than some sort of mysterious soluble substance that just happened to be associated with microbes. In today’s episode, we will see how Eduard Buchner definitely demonstrated that fermentation was caused by a soluble substance that was associated with microbes, and no microbes are actually needed.

Better still…they’re both right. “Wait…what?” Read on, O Seeker of Microbiological Knowledge, and be enlightened by this month’s entry: Continue reading Über alkoholische Gärung ohne Hefezellen