 It's time to start, I guess. So there were seven epidemics, actually, of cholera. Pandemics are worldwide. And a couple of things I want to say about the 19th century before I embark on this, it was the two greatest achievements, in my opinion, were antisepsis. And the development of public health measures, those two things saved more lives than anything else that was accomplished in the 19th century that I can think of. Maybe someone else can argue with me on that. But I'm going to lead through that. And then that'll be at the threshold of actual antibiotics that we'll discuss in another talk. So at any rate, one of the things about the World Adventure for the Soldiers, young men with, there weren't females in the military to speak of in those days that were traveling abroad. And the British Empire kept a huge army of international people. And soldiers moving from place to place acquire illnesses and spread them. They actually act as vectors. And cholera was no exception. In India, the first pandemic of cholera broke out. And again, this is a day in H1. Nobody knew what was causing this. No one had any clue. It's just it would hit and people would start dying. But they had most certainly massive mortality in India. But it spread at 10,000 deaths among British troops and hundreds of thousands at least across India. But 100,000 died in Java alone. But it burns itself out. Well, that's as I think it's called Marismas, this idea that bad air caused disease was prevalent in those days. But a banned pathogen will kill its host. A more symbiotic and better adapted pathogen uses its host like an uber or like a restaurant and doesn't kill it, just feasts on it and travels with it. And so cholera wasn't a very good pathogen. People died of it too readily and too quickly. So pandemics, terrible as they are, tended to last only so long. And second epidemic came pretty soon after that, 1829 and lasted for 20 years, really. Started in India again and went into Russia, Finland, and Poland. In England in 1831, 22,000 people. This was a huge mortality. And there were other things like potato famine. And it is malnutrition never helps if you've got a pandemic. But I've got some other details there about it. But one of the really interesting things that came out of this time, there was an Irish physician from Limerick who was a world traveler in a way. He ended up in India involved in telegraphs and trying to make a telegraph company run a line from to Europe and even did some innovative stuff there. But he studied chemistry and he looked at what was happening with cholera and people with cholera. One of the things these doctors, if you've ever seen pole dark, the early 19th century and before physicians, they did bloodletting, application of leeches, and cathartics and medics make you throw up. And toxins and they got pretty poor results. And this fellow, O'Shaughnessy, had an idea which was pretty insightful of restore blood to natural specific gravity. And his idea was to try to make saline solution and with minerals and bicarbonate and such and inject it. And he wrote and promoted this. And a physician, Thomas Lada in Britain, accompanied by two other physicians during the color epidemic actually tried this. I've talked to people that are a bit older than I who remembered making their own IV solutions when they were in residency in hospitals in the United States. A lot of aspects that almost traced back to the 19th century were still used in medical training, paid into later 20th century in the United States at least. But at any rate, they formulated a solution. They used, I guess, clean water and salts, probably not sterile water, which is a concern. But that may be tolerable depending on how clean the water was or if they boiled it or at least filtered it. And they probably had a hypotonic solution. If you have a hypotonic solution, meaning the molecules per unit volume are less than what extracellular fluid in the plasma is, then the red blood cells will take in fluid until they lyse or break open. So you can get a hemolytic effect. But at any rate, they had this extraordinary story they told and reported where they had this one female patient and she was at death's door. They called this the gray plague because they looked grayish in color as opposed to the black plague or the bubonic or pneumonic plague. But at any rate, which are the same but different forms of the Yersinia that went across Europe earlier on, starting, I think, in October of 1347 was when a ship docked in Messina, Italy, and was carrying priorettes that had fleas that had Yersinia that's associated with the black plague that had come from the Black Sea. And in five years, about a quarter of the population of Europe was dead from that. There's a lot of risk in starting to cross geographical lines and be exposed or expose others. But at any rate, they had this patient. She was at death's door and just about to die, basically comatose. And they had some kind of a needle that they had hollow needle so they could administer these fluids. And they injected her. And they injected her again several times, which was bold. But I guess this is a desperate measure. And by God, she woke up and started talking to him. And she said she felt a bit tired. And they reported this. And there was outrage in the medical community across Europe that they would be so violating the precious bodily fluids, invasively injecting fluids into the sacred temple of the human body and all these kinds of thinking points. And so that was it. Nobody else really used saline to speak of until around 1902 when physiology had come to be much better understood. And it was also after the germ theory had really gotten developed. But that, so I think 1831, I believe, was 1832, was the first attempt to give IV saline. So anyway, there was the next cholera epidemic. It was really deadly and really hurt Great Britain. And 23,000 people died there. And the reason I'm pointing it out in particular is because it sparked John Snow, who was a physician to systematically study the distribution of cases. And then he started trying to figure out the source of this malady and looked at the water they were drinking. And again, this was when people were thinking, that's what they say is from the fumes that they smell. And for one thing, the Thames and the city of London had about 2 million people. And so you don't get this by watching these costume dramas from the Victorian age or Jane Eyre or something like this. But the odiferous quality of dense populations prior to good hygiene and public health must have been pretty horrific. At any rate, he figured out that there was a whole cluster of cases near this one water source. And he got them to take off the pump handle. And the cases dropped, new cases dropped. And there we go. During this same epidemic in Florence, there was an academician who was an animus and a sectionist as he was, like a pathologist in skills. He was also good at microscopy. But he actually did necroscopies or autopsies on people that had just succumbed to cholera. And this is really a feat. If you ever look at slides and look at all the variety of things you might see in a mixed source, he identified these little comma-shaped organisms, which were the source of the disease. He called it Vibrio. And I'll show you some more of those in a moment. He published a paper on it in 1854. But he was in Florence, which was not the center of medical innovation. And he was ignored until 1865, 82 years after he had died. International Nomenclature Committee adopted the title Vibrio cholera acini as the correct name for the cholera-causing organism. So I see I've got this on my nose. 18 Robert Koch, somewhere here I have the year. Oh, here we are. 1883 Koch went on an expedition. He was a German pathologist and one of the big three in terms of the development of modern bacteriology. He went to Egypt with a group of scientists. And the cholera epidemic at that time burned out. And then he went to Persia and then on to India. And he was able to identify the same organism and publish it. And so for a long time, Robert Koch was given credit for having discovered the cholera axillus. And oh shoot, I moved myself rather than the screen. Excuse me a second. I've got to line up my screens so I can see what I'm doing. OK, here's a slide. This is Creative Commons. So I didn't steal it. And that's what the cholera bacteria look like. And they have little flagellae or motile appendages that use energy to drive it on their exterior back, so to speak, of the cholera axillus. But axillus meaning they're sort of rod shaped but curved. And so at any rate, this is again another example how so many things are discovered in so many places by so many different people. And it's often a toss up who gets credit. And overall, maybe it doesn't matter because it matters that a lot of people are trying and there's a culture that will promote spirit of discovery. And it's being carried out by men and women. But exactly who gets their name on a postage stamp doesn't matter in the big picture, really. Maybe people in the ancestry.com would like to know that or something, but who cares otherwise. It's a facultative organ, anaerobic, versus aerobic. Aerobic bacteria require the presence of oxygen and die without it. Anaerobes can survive without oxygen. The original bacteria on earth were anaerobes. And if it's facultative, like often E. coli is facultative, meaning it can tolerate oxygen that maybe prefers to be in low oxygen atmosphere. That's one characteristic about it. And pilli are these little bumps on it with they're covered with molecules that help them adhere to their target, whatever it is that they use to gain the environment that lets them proliferate. The reason they cause disease is because of a toxin, cholera gene, which is heat labile, meaning if you had water from a stream where there's a pandemic of cholera, you boil it, you'll be probably OK. This is a scanning electron microscope, microscopic picture of it. I don't have a slide on it that I want to mention. One other person, there are three people that are associated with modern bacteriology or microbiology. And that's Louis Pasteur. Everyone really knows Robert Koch, Robert Koch, who fewer people know of. But he's still well known among anybody that's ever studied pathology and microbiology. And then the one that is pretty much overlooked and named Ferdinand Julius Kohn, COHN. He was from Germany, Breslau. And he's one of the founders of modern bacteriology, microbiology, because he developed noceologic. Remember in one of my earlier talks, if you heard it, noceologic work is doing medical classifications. That's actually a big part of what is required. You've got to sort out what you got, and so you know what you're talking about, and have agreed on standards as to identification. So he classified bacteria into four groups based on shape, as spheroids or sphericles, short rods, threads, and spirals. And in a sense, those are still in use today. A spiral would be like the spirochete that was on my opening slide, which syphilis is a spirochete like that, a trepanema that has kind of a central rod, and it spirals around it. And so he also, well, I don't get too much into details that are not so pertinent to this. But he studied transformation of certain, Vic, that's a good question. Vibrio, one would think that is likely that the flagella certainly move. I'm not sure that you could see the flagella easily with a light microscope unless you used oil immersion, and it would be a bad specimen if you have to use oil immersion. Use oil immersion, you can get up to 1,200 or so magnification without starting to get prism effects or fraction problems. But maybe because they were moving around in fluid, that would make sense. I don't know specifically how he chose that term. But at any rate, I'd like you to remember it's like these were three pioneers, Pasteur Koch, Kahn. Kahn was Jewish, and they didn't allow him to take his exams in Braslau. He had to go to Berlin to his degree. And his father had bought him a really high quality microscope, university, or better quality. He used that his whole life out of discoveries. So he also established the use of sterile culture mediums, which is something else I'm going to talk about for a few minutes. At any rate, this map here shows a technique that John Snow used to sort out the distribution of cholera. So they began to realize how the drinking water might be a problem. Not everybody believed any of these discoveries. There were all of these people that are doubters and are just unable to drop the things they had been taught when they were young. This is reminiscent. Well, the whole idea of Marismas went back to Galen, who was a 200 or so common era was Roman. He died in Rome anyway, the rest of Turkey. It was part of the Roman Empire. But physicians in the 19th century, they were educated and proud of it and citing Galen. And as though that was the scripture and not able to think clearly. One other man that I thought was kind of interesting, I just wanted to mention him is William Farr, who was a British epidemiologist, a physician as well, a founder of medical statistics. And he developed a gnosology. I think that I mentioned the word gnosologic. I think that's a great word, thinking about things medical. There's a thing called the international classification of diseases and coding based on that. And that dates back to him. Oftentimes, physicians are more linguistic than mathematical. I think I was more of an anomaly. I prefer mathematical spatial thinking over language usage came to me more slowly. Then mathematics did. But at any rate, he was quite a bright guy and worth remembering. Now, there's a big thing that happened which is a fascinating story. This is from Punch magazine. It's a cartoon. It's public domain because it's from 1858. And if they don't like it, so sue me, at any rate. The silent highwaymen, they called the Thames the great stink. The whole 2 million population of London and the vicinity were using the Thames as a sewer, an open sewer. And it's miasma. I guess I was calling it morasma. Morasma is something different. Miasma is the term for this foul air causing disease. They had to shut down the parliament at one time. Parliament was right on the Thames. And they had young men waving fans by the window to try to keep the fumes out. And I think they had a hot summer which didn't help. And so at any rate, the fact that everybody's excrement was going in, their drinking water didn't help. They were bothered by the smell, but they didn't know that the problem was germs. They used lime just like they would use on bodies. Try to take the stench of a decomposing body away. That didn't help. So they passed a bill in the 2nd of August, 1858. A stewar, as I understand, lost three daughters in childhood to typhoid salmonella, food poisoning. Said, can be invasive and cause liver abscesses and all kinds of things. So this kind of public health progress changed the world. Human beings take this for granted these days, but it was one of the massive accomplishments of the 19th century to have building projects like this, have cooperation, and focus construction projects, and also the scientific work to back it up. It was like 1862 when a stewar published on the germ theory of disease. And basically, that's in microbiology. It's of importance comparable to the atomic theory that all substances are composed of tiny particles that are atoms or atoms combined in bonds that make molecules. And people didn't believe that even up to the time of Boltzmann committed suicide thinking all was lost because he had based his entire beliefs in physics and his entire career on the atomic theory of matter and developing statistical mechanics. So at any rate, the germ theory is a big deal. But it was not until the late 1890s, so like 35 years before physicians in North America really believed it. They rejected it saying it sounded like science fiction or just made up. They didn't believe that these little specs could cause disease. And they just thought it was nonsense. And to that, I have to ask, what is it about North America that makes them so they choose to be behind on science? Anyway, my climate change. So it was a fifth epidemic. It was terrible for Asia, Africa, South America. France and Germany had got hit, like I mentioned earlier, Hamburg. And Russia got hit hard. And the development in Russia was not as far along as it was in Western Europe. And Japan also got hit hard. But the quarantine measures of John Snow really had made a difference. And a vaccine was developed for cholera in 1892. So as to mention Koch, I think in later or four later talks, I'll have a picture of Pasteur for some of the things he did. But Koch was credited with, let me see here, anthrax and cholera and tuberculosis in terms of discovery. And he actually promoted the germ theory of the disease. And as Koch postulates that the germs always have to be found. It's very intuitive if you have the idea of germs causing disease. That if you have an identifiable disease to attribute it to a specific bacteria as a causative agent, you have to be able to demonstrate their presence all the time. And that you can culture it. Now, culturing, that's another whole big step that I want to talk about for a minute. But just, and it's another aspect of how, if you think of it, the development of culturing bacteria relates to agriculture and then food science and preservation of food. And at any rate, after you've isolated the bacteria you think is accountable, you have to be able to culture it. And then if you introduce it into an organism, actually an animal that's susceptible, you would induce the disease. And for something like anthrax, this was easier. I guess 1870 was when he discovered the anthrax as a cause of these. Basically, it has an exotoxin. It's a complex exotoxin, which is various combinations of like three protein units. And it kills cells. It can start. It's a horrible disease. I may recall some years ago, people were mailing powder to people that had weaponized anthrax. In that case, anthrax is sporulating. It forms spores. And if you inhale it, the spores are tiny. They first go in your lungs. They don't make the lungs influx. They're picked up by macrophages, these spores, and carried to lymph nodes in the middle of the chest called the metastinum. And there, the spores convert. And because they're in a nice little susceptible area and start to take over and cause mediastinitis. And then that's pretty sick. Mediastinitis, that used to have a very, very high mortality. From there, it would then infect the lungs, kind of reverse. Oftentimes, you have something in the lungs that causes mediastinitis. But there, it goes from spores getting by your friendly macrophages, which are inflammatory cells that eat up foreign maters. And they take them behind the walls of Troy. And there, they crawl out of the horse and spore and start to do destruction. And then they go to the lungs. Once you get into the lungs, pneumonitis, a second phase of this kind of pulmonary anthrax, the mortality is about 85%. Most of the time, anthrax is on the skin. Hunters and people to handle animals would get it. And sometimes it would clear up, and sometimes it wouldn't. It had 30% to 50% mortality, I think. It was originally called woolsorter's disease for laborers who would work with wool cut from sheep. Sheep may have anthrax. I come from old people who came from old people. And my father told me once about, I remember, and I once had a brush and a cup when soap in the bottom. And I could soap up the end of the brush and then put the soap on my face for shaving. And my father told me about they used to make these brushes out of bristles of livestock and cattle or horses. And if they had anthrax, there were cases where people use those brushes on their face and they get a little nick, and they'd get anthrax on their face from the brush. So cutaneous and pulmonary anthrax and intestinal anthrax is another major presentation where that's one way cattle get it. Yes, anthrax is a terrible danger. It's used in terrorism and as a bio weapon. But cattle eating grass, they could be eating where some cattle had anthrax and were buried years before. And Pasteur showed that, yes, especially if those spores last for years, and earthworms can ingest them and carry them to the surface and excrete them. And the anthrax spores get taken up by the grass. And so cattle eating the grass ingests the spores. So disposal of animals that have anthraxes is pretty tricky. If you bury them, it has to be really deep. And the fourth way you can get it is by injection. Of course, Pasteur developed a vaccine for anthrax. And then he injected. He had a sheep that had been vaccinated. And he had sheep that had anthrax. And he blood from the anthrax injected sheep and injected the ones with the vaccine. And they didn't get sick. It was the ones that were naive or unvaccinated. Their immune system had no previous knowledge of anthrax. And they would get sick and die. So at any rate, again, a lot of what is considered medical advances done for the sake of agriculture and food production, I just think they're really tied together. That's one of the themes I just wanted to make in this talk. So I should go. It looked like I'm doing OK in time. So there was yet another cholera epidemic at the end of the century, which again went into Russia and parts of Europe, Northern Africa and Middle East. By 1923, it was pretty much limited just to outbreaks in India until around 1961. And there's been kind of a continued slow burn of cholera since then. And they can tell what strain it is and this sort of thing. They have vaccinations. I was in Spain. And there was an outbreak some years ago. And I didn't have my card. So it seems to me I told this last week. They had an air gun. I went to Spain. I had to be able to show I had been vaccinated for cholera. And so it basically shot the dosage through my skin. And I don't think I even got a band-aid. I wasn't too worried about it. I did tell this before, I think, because I was so told about how in World War I they would use the same needle going down row to vaccinate fruits in the United States. Now, they're gone now, basically, those soldiers. But if you talk to them, they'd say, man, you didn't want to be the last in the line because the needle was really dull. So you really had to jab it to get it through the skin. And I was sort of stunned by hearing that. But at any rate, in, no, it's, well, it's North America. And it was, Abe Flexner was a man who went and studied European training of doctors. And he came back and basically in the United States, for the most part, doctors were trained by apprenticeships. And he proposed that they have basic science training and then in anatomy and biochemistry or chemistry and organic chemistry. And I'm not sure when biochemistry became self-aware and started calling self-biochemistry. But embryology and basic biology that's pertinent to medicine followed by a year preclinical pathology and how diseases are caused and effects on physiology and how to diagnose them and then have clinical training in hospitals, training hospitals. And because of his work, it was 1916 when I'm sort of going on beating from years ago. But 1916 was when the average patient in the United States could go to the average doctor and have a better than 50-50 chance of walking out better than when they went in. So there was, there's been a lot of range. Yeah, flip a coin. Well, it's better than that these days, I think. But it's taken a lot to do that. So there was this earthquake in Haiti a few years ago. And Nepalese volunteers apparently contaminated the water by their excrement. They carried cholera. And because you can't acquire immunity if you survive it. And you can survive it if you get fluids. Basically, you die of hypovolemia or loss of body fluids and shock and your kidneys shut down. You have adequate blood flow to your kidneys and they just die. And you die of renal failure and dehydration and heart gives out and it goes pretty quickly. But at any rate, they were able to look at the strain of cholera and it matched with what would have come from Nepal. And they're still struggling with cholera there in Haiti. They didn't have that before. They had plenty of other problems. So let's see. So we're still struggling with cholera in different places. It was recently 1994. There was a refugee camp with about a million Rwandans. And 12,000 died of drug-resistant vibrio-cholera strains. So universal public health is really what's needed where safe drinking water and proper disposal of human excrement, as that's a fact of life, will do more to protect infant mortality and things like that. And so it's considered that there's this basically ongoing smoldering seventh cholera epidemic. And somebody mentioned Graham as a leader in bacteriology. But I don't think he's at that level. But he did make a pretty outstanding contribution. And this reflects the emergence of organic dyes that were, I think it was 1852, trying to think of the guy's name who the first organic dye on London at a little laboratory in his apartment, blocking on his name at the moment. But at any rate, using organic dyes, you could have tissues that were thinly cut or a smear of germs or material that you wanted to see what was in it and look for germs. You could put that dye on it. Trouble is the dye would soak on everything. So Graham developed this technique. And it resulted in a dichotomy that applies to most bacteria, Graham positive or Graham negative. The Graham positive bacteria, the ones that are really susceptible to psoins, they have this little glycan cell wall that's in the top there in red. And then there's a tiny space. And then there's some membrane inside. And Perkin, yes. Thank you. William Perkin. And the Graham negative has a really sparse peptidoglycan layer. And it has an external outer membrane, which is lipopolysaccharide. And it does not take up the purple stain that's used in the Graham staining so readily. Let me tell you the technique just in a nutshell. It's not hard. I've done these before. And it's really pretty cool. I've got a bunch of microscopes. I've thought about setting it up so I could do Graham stains. And I haven't done it, but I should. At any rate, you have crystal violet. You take a smear, and you need to do something so it doesn't just wash off. So usually you just quick heat it. And so it applies. You can also put some alcohol in it, let it dry, ethanol. But I've always just heated it. You don't heat it till it's that hot. Just to warm it up. You can do that over a flame. And then you apply the crystal violet. The trouble is that it won't adhere to the acids, like nucleic acids that it likes to adhere to, and peptidoglycan cell walls, unless you have a mordant, something to make it stick. So you apply a drop of iodide solution. And that traps the crystal violet in a combination with target molecules. And then you dip it in ethanol. And then you counter stain with a red stain, like saphenin or basic fushion stains are started as better for gram negatives. And it does make it incredibly easier to see what you're dealing with. And if you see bacteria and you are describing their morphology or their shape, spherical or spiral or axilla or whatever, and you can stain them with gram stain, there are a lot of other stains these days as well. You can further classify them. Gram positive, gram negative makes a big difference. For instance, like strep or staph would be gram positive. Anisepsis, I'd mentioned, is a big deal for the 19th century. And Joseph Lister used polyc acid, which is actually phenol, which is most easily done as benzene from petroleum. And then you can sulfinate it or act it with a chloride at one of the carbons. Benzene is a six carbon aromatic flat planar ring that resonates. And so it's pretty stable that you replace the, I think you put the chlorobenzene with just one chlorine molecule with a fly or soda ash or something that's pretty strong and heated. And then you'll end up with a phenol with an OH or hydroxyl group. That's what carbolic acid is. And Louis Pasteur used carbolic acid, which is phenol, and on anthrax to attenuate or kill the anthrax so that you can use it to make a vaccine. If phenol is pretty reactive, if you ever touch it to your skin, you'll get a burn. So there have been times I've used actually high concentration phenol for certain things. So Lister was the first to really use antiseptic techniques. And he persisted and got attention for it and eventually became Queen Victoria's personal physician. He used phenol solution on dressings on this young patient who had a leg injury that often would end up in amputation. At the turn of the century, 1800, the number one treatment for limb infection from trauma, especially, was amputation. And often 40% of the people that got amputations died. It was pretty rough, but that was the best they had. They didn't know what they were dealing with until 1862 when Pasteur and Kahn and Robert Koch really promoted the germ theory of biology and medicine. So at any rate, all this led to eventually having areas and hospitals that are considered germ-free that gets disinfected after every patient has left for whatever they're in an operating room. And so it's brought down surgical infections enormously, along with some other things. I wanted to point out one other thing about antisepsis. If you're out in the frontier or the wilderness and you've got an infection, if you've got wine or vinegar, you can use that to help your dressings not get infected. You don't have sterile stuff to work with. I had some notes here. Oh, Hippocrates mixed wine and vinegar in dressings of wounds, around 400 CE. And I suppose somebody figured it was a salad. Well, wine and vinegar would not have the oil. It was a lame joke, I'm sorry. Wine was used in dressings in 14th century Italy. Sir John Pringle coined the word antisepsis in an article he wrote in 1750 on experiments upon septic and antiseptic substances. And later mercury, 1766, mercury was used as an antiseptic. And I think in Beethoven's apartment, there was mercury or probably syphilitic lesions he was dealing with. 1811 was when iodine was first used on treating wounds. And it's not something you should take. Internally, it's pretty toxic. I think it's 53 atomic weight, atomic number 53, and it's pretty heavy. So it is given internally in the form that it will show up in the bladder. And when they started doing X-rays, radio contrast effects in the bladder associated with iodine. And so iodide in contrast agents arose from that. In addition, this first surgery there with an anesthetic was done in 1848. In 1884, this advance on the steam digester, autoclave, was invented. And it was used not only for sterilization, but also for vulcanization of rubber. And Ulstead was the first to use rubber surgical gloves in the United States. And there's a couple of others there. It was actually around 1962 that the first latex gloves that were treated with gamma radiation were used in surgery. And they were powdered, and powder can cause allergic reactions and fibrosis and severe inflammatory response. And I was always bothered by the powder when I did surgery. So I thought it was nuts, I suppose. But I always had them bring a basin. It seemed like it bothered my hands too, but it would make my hands break down. But they were powdered on the outside as well. So I would wash my blubbed hands after I'd gowned for surgery and in a basin of water, a sterile basin, sterile water, just to get the powder off. And in 2016, I think, all powdered gloves were banned in the United States of the FDA. There was, let's see, one other... Oh, 1795, there was a trenches on the endemic of the Puerporal Fever of Aberdeen, way up north in Scotland. There were all these women having Puerporal Fever. And Alexander Gordon started requiring everyone in that hospital to wash their hands. Later, around 1942, I think, where he had some of ice in a... It was Hungarian, but he was working in Vienna. Notice that the kidwives had less post-livery fevers in the women and less maternal death than the doctors did. And they had no sense at the suggestion that they were spreading this infection and causing harm. But that was a historic point. It's often pointed to the improvement of sterility in hospitals. Also, in 1883, Gustave Neuber of Kiel was the first to use a sterilized surgical gown. So, basically, in the last part of the 19th century, more and more steps were taken to prevent the participants in surgery, whether it were nurses or technicians or anybody giving anesthesia or whatever, from being vectors in disease. And you're right, it's a lot of people suffered terribly in the Civil War, and deaths due to drinking bad water. I want to tell one last story, if I may, just over. Do I have time for one last story? Okay, thank you. And there was a search for something to grow bacteria on. Just to be able to culture bacteria was a challenge and required the input of a lot of people. Pasteur used a broth and made from meat or vegetable matter. I'm sorry, I bumped my microphone. And Robert Koch wanted a solid culture medium, and he actually tried to use slices of potato. I bet that smelled rich after a day or so when he would inoculate it with bacteria. In 1882, after not being able to use gelatin, because it would be liquid, you want to culture 35 to 37 degrees, sonograte or so, similar to body temperature. And gelatin tends to liquefy, and also it gets digested by bacteria. So instead of having a solid matrix where you can see bacteria growing in colonies, which is what they wanted to... Well, eggs are also kind of liquefy, unless you heated them and cooked them, and then they're coagulated. Eggs embryos, as embryos are used for cultivating viruses, but at any rate, in 1882, there was a brilliant suggestion by the frau of Walter Hesse, who was an assistant in Robert Koch's laboratory, and her name was Fanny Hesse. And she suggested that they use agar. Agar comes from red algae, and it's basically got no protein, no fat, no sugar. It's very fibrous, and it's powdered, and it's a complex algae, like seaweed. And it had been used in making desserts and Asian dishes for a long time. And it's a vegetarian alternative to gelatin, if you don't like eating bulbs of horses and cattle from broken down cartilage. But at any rate, it's not hard to make a culture medium. It's about 5.2 gram percent, meaning 5.2 grams of powdered agar, that's sterile, and you measure that out and put it in with a total of 100 milliliters of deionized water. And at room temperature, you can do it in steps and mix more water in until you need to get the powder to mix in with the water. Now if you took agar and just ate it straight, solid, it probably would obstruct your GI tract, even in the esophagus, because it swells over 20 times its size. It takes up 20 times its volume in water. So it's sort of like peat, peat moss. But at any rate, so you make this agar and mixed in with 5.2 grams and 100 milliliters of deionized water and get it stirred up really well, put it on it or something and all to clave it. And the, oh, I think it's like 120 degrees, so centigrade, and for up to 20 minutes or so, and then on the average petri plate, you can put about 20 milliliters and let it solidify, and then you store the petri plates upside down. Petri plate was invented in 1887 by Julius Petri, TRI. And basically there are two lids, one slightly larger than the other with glass, which was one of the great discoveries of mankind. And they nest, but one over the other is a lid. And when you got a solidified agar in there, you store it upside down, so the moisture doesn't tend to collect on the agar plate to be dry. If you wanted to culture something, you can make a little wire loop, take any wire and make a little loop and then heat it till it's red hot, get a little black body radiation and then sample, get your sample just a little bit, get it with a little water if you want, and basically before you get your sample, after you've sterilized your loop, you probably need to cool it down waiting or else you can just touch it to the agar in one spot. And then you get your sample on your little loop and then you just scoot it over the surface, like in the zigzag of the agar and then you put your agar in an incubator at about the same temperature as my yogurt maker and at 35 to 37 degrees centigrade. And you'll see colonies of bacteria, pure colonies grow if you have looted your sample properly. And since around 1919, there was a man, Edward Rosenau, who combined dextrose broth and calf brain and this led to a form of agar called brain-hard infusion agar, VHI agar. And it's particularly good. We used it for cultivating streptococcus but also for mycology. If you're one of culture fungi, it's a good agar for that. A lot of these organisms are very fastidious and some can't be cultured at all. The syphilis, for instance, nobody's been able to culture and appear short of a cell culture. And that's basically it. I had one last quickie point I wanted to make, somebody I wanted to mention and that was Harvey Washington Wiley who you've never heard of but he was a physician who was sort of a self-educated chemist and he ended up on the faculty at Purdue University which was a land grant college started based on a law signed in by Abraham Lincoln and he made it his purpose in life to have unadulterated food and it's a long story but it would really be something you might enjoy reading about. Harvey Washington Wiley, W-I-L-E-Y his work led to the founding of the Pure Food and Drug Act of 1906 which led to the FDA, the Federal Food Administration and after he had had that he started working on purity of pharmaceuticals because there was a lot of problems with adulteration and things not being what they're supposed to be. So again, a lot of people of conscience and of responsible intelligence are to account for the fact that you live a life that's free of miseries that are avoidable and millions of people really have contributed to all this body of technology and culture of being able to assess the microscopic world and understand how it's impacting us and how to keep it from hurting us and recently they've been attacking the FDA they wanted to move it to Kansas City basically to get rid of researchers because they want to control the message and some of the researchers they were concerned about climate change and the FDA is part of the Department of Agriculture, you see. So, well, that's it. I'll stop right there and I've gone 13 minutes over but food for thought. That's having mentioned, I'll show you my last slide here which is just kind of a statement of some of the big things I think that have come together for all these things that humans are able to do in the coordinated, educated fashion and if this is all that matters to you there are jobs, jobs, jobs involved in all these activities. So, thank you, Mike. Any questions? Thank you, Dave. Thank you, Mike. Thanks, Edgar. Thank you, Tata. Thank you, Cass. Thank you, Katja. Get a dessert with agar, Cass. Thank you, Vic. I found it as I was reading and planning and what I would say. I just thought I got to mention this and I got to mention that. It was a fun topic for me. I hope it was fun for you. So, perhaps sometime next year I could talk... Oh, Symbol Weiss. Well, he got people washing their hands. There's a lot written about him. I had a Hungarian friend and colleague in Philadelphia who was just so proud of him. But basically the take-home story from him was that washing hands between cases. The thing was they had women that, if they got the streptococcus, which is an invasive gram-positive cacus bacterium and can cause abscesses and purulence, and they would go from one delivery to the other. And this was the time, you know, and there was one article I read that was an anesthesia history review. But like 1850, the chances were not very high that you would go get a surgical procedure and have the doctors wearing anything but street clothes. And they were proud of it. They were proud that they could go in there, find clothing, and take off a leg and not mess up their clothing. It must have been the same with deliveries. And they would go from one delivery to another without washing their hands, so they were direct vectors, transferring this pus-forming or virulent streptococcus from one patient to another and postpartum. The mother would get streptococco infection in the raw lining of the uterus and get sepsis and die in any cases. So it was really kind of a horrible situation. Again, it's interesting that I think it was 1795 in Aberdeen that Alexander Gordon noted the same thing. But it takes quite a while for even the educated public, target public that you want to get the message to come across and buy it and to change their culture. That's about all I know to say about Simmelweis at the moment. Thank you. Any other questions or comments that I have here? I got turned around in the talk. So my avatar was speaking with its back turned to you. My next talk, I wanted to continue on this, but focus on mycobacterial diseases like leprosy and tuberculosis called mycobacteria, like bovine, tuberculosis and that sort of thing. And malaria and syphilis. On those three matters, I could do that in one hour. Well, Lyme's is kind of interesting and it's related to a spirochete. That may deserve a talk of its own. I wash my hands quite a bit. On the other hand, I'm not uncomfortable about living in a sea of germs. And one of the talks is I developed more information about antibiotics. I want to tell you about Rene Dubois, I guess his family, he was American born in France, but his family actually pronounced Dubois, which is a sort of atypical pronunciation, but he was kind of revised the germ theory to take us away from absolute fear and dread and loathing of germs to understanding that living in a sea of germs and to get disease from them is more the exception. So he played a role that was pretty significant in the development of antibiotics. Well, thank you for attending, Mike. I appreciate it. Yeah, and if you have to wash your hands a whole lot, I don't really recommend using these antiseptic soaps that have harsh chemicals. I think getting some suds, the suds disrupts viral capsules and bacteria and you don't have to wash in real hot water either. It removes the oils from your skin and you can wash with cold water and your hands will hold up better. Thank you, Shanta. Washing with pure water only while rubbing your hands thoroughly and thinking under your fingernails and letting it wash under the fingernails is better than no soap or water at all. A little bit of suds helps with getting rid of pathogens, especially if you're going to do food handling. A brush to wash hands. You know, in surgery, you use a brush and you especially brush your nail beds. The fingernail beds and underneath the nails is an issue for a lot of bacteria.