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00:01 Hmm. Mhm. Mhm. Okay . Um Welcome. Right,

00:33 This is making creases here. Hello? Oh, that would

00:51 There we go. That did Okay. Alright, fantastic.

01:03 That's pretty good. All right, to the Super Bowl edition of Micro

01:12 . Right? There will be a bowl themed party clicker slide when you

01:19 that, you know, that's we're . That's the last slide we're gonna

01:23 today. Okay, so until so I know that I mentioned last

01:29 about I changed it on blackboard, the schedules on blackboard, but I

01:34 actually change the natural syllabus. You have printed out where it said chapter

01:39 was a flipped class. It's not not so it's gonna be usual

01:45 but the next week, so next uh right here in front of me

01:52 . Okay, This one. That that will be in europe

01:58 Is open for those of you that on, everything is available in that

02:06 . Uh flipped the folder with the class stuff. Uh in there is

02:14 chapter six part one video lecture. that's all available to you.

02:20 Um so I know, so unit is three chapters 6, 13

02:31 Okay, so but we were about next week. So that's next week

02:37 today. We start unit two. . Um of course exam is next

02:47 and saturday. Uh so what And so what we got, you

02:52 , one quiz opens tomorrow. That's comprehensive. It's like 25 questions or

03:00 . Um covering one through 134 and of five that we'll talk about

03:08 And uh that's it. That's smart stuff is do you remember this?

03:13 uh this has kind of come not to back but Day in between.

03:21 that Chapter five is due on So most of the smart stuff do

03:25 Mondays, but there are a couple are due like in the middle of

03:28 week. Typically these are ones that occurring before an exam. So you

03:33 want a couple of days before the to a question about it. So

03:37 why that's due on Wednesday. Um Alright so we're gonna finish part

03:44 of chapter four and then a little about control of the vehicle. We'll

03:48 up the rest on Tuesday. And uh All right. Um Tuesday's elected

03:56 gonna be 40 45 minutes or Okay so we have that, we

04:02 have as much to finish up and next week. Okay. Um Any

04:08 before we roll along here? So I just want to begin with

04:15 a recap of what we talked about time? Okay let me get this

04:18 of the way. So um so talked about the fast growth current,

04:25 ? We have different phases uh log death. Remember that? These can

04:33 right? Like phase can vary different factors that can influence that everything from

04:42 nature of the growth medium, the is growing on and what is going

04:47 uh you know, slight temperature ph differences as well. There are

04:52 differences. These can all kind of a situation that has to get activated

04:57 . Right? It's not gonna pop in and start growing. It's gonna

05:00 to adjust, maybe has to has turn on certain genes, turn off

05:06 genes adjust the temperature and ph and all those things factor into time.

05:12 , what does start growing? Of it can grow very rapidly.

05:17 so we have all these kind of a baseline but of course these can

05:24 can vary in terms of lag the inflection of log growth, right?

05:30 vary the the onset of stationary phase quicker. So you know, these

05:38 will all occur. It's just that duration the rate can differ depending on

05:47 number of factors to my program the growth medium, other conditions

05:53 Okay, so then we did a of growth calculation problems. Right?

05:58 this is uh equations And all these are given to you, memorize these

06:04 . So there's a couple at most problems. Um but you'll have the

06:10 there that you'll need to solve. , and you can bring a handheld

06:16 , you can use your cell phone you can use a handheld tactic and

06:20 casa will be made aware of So people in the progress that will

06:26 have to you about having capital, they'll know ahead of time that this

06:30 a lot. Um So so in generation time, so we use that

06:37 different types of problems the generation Uh the Hawaiian does it take to

06:43 generations of the population? So again goes through those practice problems if you

06:49 to if you have questions let me uh beyond just the bad growth where

06:55 they're not other than taking other than and taking samples out to measure

07:02 You're not doing anything else. So just basically letting it go to the

07:06 and whatever the cell density is which represented by maximum cell densities here in

07:13 example whatever that is is what it right? You're not trying to um

07:19 it anymore. But you can do of course when you do things like

07:24 , that truck, right, add part of it typically to boost

07:31 Right? So obviously your goal in this just to get a lot more

07:35 right for whatever purpose. Okay then course you can really control things in

07:40 bioreactor where you know all the various the different parameters of growth, oxygen

07:48 , oxygen um ph um nutrient All these things can be in temperature

07:56 all these things are you're controlling all things and so when you do that

08:01 course the culture very okay. And what you do when you're doing kind

08:08 commercial steel. Uh um you can't this class. But, you

08:18 of course, what were you working ? And if you're in a lab

08:22 and you're doing this kind of resources may be limited. Maybe you

08:29 have one of these fancy computer controlled reactors, but maybe you don't.

08:35 . So, do you still have there if you only have shaped

08:39 Yeah, you do. You can can um control it to a

08:44 It's much more of a pain in butt, right? Because you have

08:47 computer helping me out, right? you getting up at intervals and looking

08:53 your shape class, for example. ? So you can do things like

08:57 see how it's blue colored, You can add ph indicators,

09:03 So, I I actually did this . So you can have like uh

09:09 forget what it's called, but it's that when it gets acid it's yellow

09:12 it turns basically reddish. So you kind of eyeball it, you have

09:16 ask them in the base and neutralize . Um You can uh of course

09:22 in an incubator while you're incubating it incubators, temperature control it that

09:27 Uh then that ph temperature, What can we do in terms of

09:33 for the shake flask? Okay, this thing? Well, you can

09:41 see these little these little indentations in bottom of the flask. Right?

09:47 four of them. Right? What you think that you think that's nothing

09:52 ? Actually make a huge difference, ? These are what we call um

09:57 don't need to write this down. this is what we call. These

10:00 baffled, baffled, baffled. I know what's going on, baffled shake

10:09 . And they always prepared this to black girl America. Ask what are

10:15 apples going to do for you anybody about it? Think about it spinning

10:22 a in a shaker spinning. What these little things he's gonna do compared

10:28 just being flat right here is the . Just visualizing what's gonna happen.

10:40 or say something. Yes. yes. Because what does do including

11:03 , that's the key. So the think the waves of the ocean,

11:07 ? Waves of the ocean. That's you mix air. That's how you

11:10 air. Typically vibrancy where waves are created. Wind, right? That

11:17 top parts of the marine environment or more options because that's where all the

11:21 is occurring, mixing in the same , the shape fast. It created

11:27 lot more turbulence more. So the thing you can do is to crank

11:33 the speed at which this thing is . Right? So those are kind

11:37 ways you can kind of um you know, add more oxygen as

11:41 cells become depleted. So again, can do all these manipulations. But

11:45 , it's much more tedious, Than um having a computer do it

11:51 you? And having all these things monitor everything real time. Right?

11:55 this you're just monitoring your eyeballs, means you have to come in and

11:59 at it. Oh, I got yellow base, right? So you

12:03 if the ph changes are occurring pretty , that's growing fast and that like

12:09 these are kind of things so you do these things in a safe

12:13 Okay. But again, yeah, probably don't wanna commit it two o'clock

12:18 the morning and every couple hours to on this thing. So if you

12:24 be in the lab that has money has bioreactors, bioreactors anyway, so

12:34 , so let's move on. Any about any of these topics. Don't

12:40 , I'm not gonna ask you what's . Okay, there's lots of stuff

12:44 throwing out because I've done this for the real world and some of you

12:50 going to encounter these things. So for you, those of you that

12:54 in that. Yeah, yeah, . So his comment relates to the

13:14 last time about if you you we do fit batch or even in

13:20 reactor, we add we add we in at certain intervals. Um

13:26 we pump in nutrients because you can't up front at the same time,

13:32 becomes a osmolarity issue becomes too hyper outside the cells. So the cells

13:38 doing water. So they're fun and problem and they don't really grow.

13:42 that's why you kind of have to these additions um while they're growing.

13:48 so you don't get this problem? . Alright so this question because this

13:53 gonna be 22 are gonna pop up the first one here. This brings

13:57 into the um the last two topics Chapter four. We're gonna talk about

14:04 we get into Chapter five. Okay this is often a nutrient urban phenomenon

14:11 by a lack or depletion of So I say often it's not the

14:33 thing that causes it but it can certainly be uh something that does cause

14:40 to occur. Okay. Okay. . 10 down 10 53.

15:25 Um Yeah. Well let's look at second question. Uh Let's see.

15:32 remember that I think um Next one . Okay. Same same choices.

15:44 answer choices. This is often a phenomenon requiring a surface and the ability

15:53 attach. And I would probably I say I would correct this by saying

16:05 they didn't make it blonde. Okay Chin. Alright so uh yeah

16:51 is that's biofilm formation. Okay. Impact. Probably good boys in that

16:58 . But it is also a nutrient . The previous one. That's endospore

17:08 . Okay. Um If I had maybe um stationary phase you might could

17:18 that could be a correct choice. because you get to be deprived

17:25 Um But what what you're given? definitely in those four formation.

17:30 Um Alright. Which which is our two things to talk about before.

17:35 , so we'll start with biofilm formation . Okay, so this going

17:43 Yes, uh biographer mentions anything, of course a surface occurring on the

17:50 and ultimately bazillions of microbes, ultimately form. Okay. And so we

18:00 that initially in what we call, say like at 12 o'clock in two

18:07 , these are typically turn micro Okay, so so it's beginning to

18:12 together and begin to grow. Uh through the 12 and three. Here

18:18 go clockwise. It's obviously on the is also occurring on the surface and

18:24 two dimensions. Then as we get uh seven o'clock nine now you see

18:34 going up so now we're going first three dimensions. So obviously this represents

18:39 lot of cell growth and if you're sustain with biofilm which obviously contains lots

18:44 sales, you're gonna have nutrients constantly supplied to them. Okay? So

18:51 you let's see uh to walk to the sec 100 auditory like walkthrough walkthrough

19:03 one walkthrough snr one as you're going Cullen, right then when you come

19:09 and go downstairs and look to your you'll see I think lately, but

19:17 see this right here, okay. used to be a bored sitting

19:23 They used to have a board and was draining out. Right? And

19:29 board was full of, it was biofilm that was occurring like they had

19:33 form in here as well, but on this wooden board was since been

19:38 away. But all this down here well, biofilm because that thing's gripping

19:45 likely gonna freeze Of course. So it's not, I'm not sure,

19:48 not sure what state it is right . But the point is this constant

19:54 grip, grip, grip of nutrient and that's what can initiate and sustain

20:00 . Okay. Uh You know there's window, this is a window on

20:05 building. Okay, you can see growth right there. Um So if

20:12 look around you can see these right? You look at your

20:15 right? You can look at your shower, you may see about

20:19 It probably means anything your bathroom if seeing that. Um You're right,

20:27 . Excuse me, while I put in my microphone here. Um Let

20:34 continue on on this thing. Um uh so again, by what's common

20:42 these 22 pictures, of course it's surface all about a surface and growth

20:49 going to occur on the surface. the state itself not only having a

20:54 nutrient supply but also a constant um ability to attach that surface.

21:04 attachment to the surface. So these two key things surface and attachment and

21:09 supply. Okay and so again biofilms fairly widespread. There's species species

21:23 Okay uh and biofilm formation is not this random you know, coming together

21:30 and just kind of all grow together a different species. And they're all

21:34 all it's highly organized orchestrated process. and coding process. Right? So

21:44 one has to occur in 2 to then four. Um So it's a

21:48 organized process. Far from me. a random thing. And so biofilms

21:55 the soil, biofilms on water. They're not all bacteria, the algae

22:02 other types of microbes. Um You from medical from a medical standpoint

22:09 Right? Capital. Right. Um tubes, um heart valves. Any

22:16 of medical devices where they provide the also can induce growth of uh pathogenic

22:24 of bio for reformers. Right? this is showing you a a biofilm

22:31 a on a capital. Okay. these medical devices right. Comes packaged

22:37 know fairly package right there. And people that handle these maybe they'll handle

22:42 property right? Get their hands on , unloaded hands on it or something

22:47 otherwise contaminated. And then that's where you can introduce one of these bathroom

22:53 types. This is and there's staphylococcus that are bad for informers. Some

22:58 . Okay. And so um so contamination can lead to or improperly uncertainty

23:06 staff is on your skin and mucous . Okay uh put your nose um

23:12 surface. And so it wouldn't be know, don't take proper care.

23:19 could be easy to actually have this . And so they begin to grow

23:24 ? And of course represents a lot thick growth that ultimately can occur.

23:29 and so again these these medical type biofilms are very problematic to get rid

23:36 , right? They respond to But in this biofilm environment you have

23:44 Micro environments in that big environment where have differences in growth and metabolism of

23:54 species in there. And some of are more resistant China box.

23:58 So it can become a problem. know when you have one of these

24:02 of infections this isn't a week or days over this. This is something

24:07 weeks and months. Okay so you you can get a hold of

24:11 Okay? Um So something to be of. And so especially those that

24:18 going into the medical profession uh your right to wake up with that sticky

24:26 in your mouth. Those are courtesy bacteria that are forming biofilm. Now

24:31 they've enhanced this okay with adding some of a dye to enhance the growth

24:37 seeing, right? But the point , you know it's plaque is due

24:41 bacteria form biofilms. Um This thing something um I have encountered previously encountered

24:51 uh fast food restaurants or any restaurant serves uh when you fill it up

25:01 ? It has to been going to uh the thing right senior 7-11 or

25:07 convenience store where you fill up your . So it'll be like a tank

25:12 that the syrup then it goes to right nozzle. Uh sio two sio

25:18 compression. If you win the working of these places, you know what

25:21 talking about. But that sugary kind tube that sugar and tube and environment

25:28 create these kinds of things that form sugar sugar polymers right? By products

25:35 their metabolism. The people that own places are kind of unaware of it

25:44 they begin to know this over And then get odors and that kind

25:48 drive people away. And that's when say we need help with this.

25:52 ? And so again, biofilm. pipes of course are very surface and

25:57 typically have something a nutrient flow going some kind of acquis flow containing nutrients

26:04 that's what can sustain the members of biofilm. Okay, so again,

26:10 you over a state this but you surface attachment and nutrients. So those

26:18 the ingredients for bio phone. Um so the process. Right?

26:25 basically look at five stages. So and it's all these chemical signals being

26:33 here. Okay. And so initially have what are called plank tonic cells

26:38 you haven't called stickers. No stickers swimmers. Right, So Pentatonix

26:46 They are kind of seeking out a surface upon which the former.

26:55 so um if there is a favorable there is a surface in a favorable

27:01 to lead to a biofilm then you attachment right? And um the swimmers

27:09 to stickers, right? So they the jelly and then february maintain

27:16 Okay, now um so in this here attachment begins to form grow a

27:24 bit. So you have what are ? I'll show you a picture here

27:26 a second. Micro coffees, small . Now, whether it goes

27:34 Okay on are there if enough cells present? So enough cells are present

27:44 those cells are throwing off this chemical ? Okay enough cells are present.

27:50 you have enough chemical signal being formed a threshold. You have to have

27:55 growth to create enough chemical signals to initiate and attract more and more cells

28:02 the surface so its threshold. And this is what's called this quorum

28:08 phenomenon. Okay. Um Well, explain in the next slide but that's

28:13 of it can kind of not occur all. Right if there's not enough

28:19 there. Okay. Now if it occur then you'll get extra place.

28:26 is basically the glue. If you . Yeah, that's the essence of

28:32 biofilm. Is this production of this supply, Saccharine can also have some

28:37 in there but it's kind of what everything. You get what they're embedding

28:42 in this in this biofilm. Okay maintaining it as if there's plenty of

28:49 everybody's fed then we grow not just two two dimensions about the girls

28:55 Right forming what's called a biofilm Okay. Of course it represents lots

29:00 lots of self self worth. But of course it could be that

29:06 know because the pipe or maybe the of nutrients is stop somehow and so

29:13 can't sustain all the members of the anymore then um then it will move

29:20 dissolve so to speak. And then that used to be stickers right?

29:27 revert to being swimmers. That's when break off and then find a more

29:31 environment and then start presumably start the again. Okay we kind of see

29:37 here. Okay so uh cells in iconic swimming slash swimming state.

29:46 Um that initially attached and you can have um the twitching motility can be

29:55 here as well so they can have kind of movement on the surface.

30:03 . And um so again I began but to get the the image.

30:17 if there is um enough presumably if enough signal that means if more and

30:24 cells are being coming to the that must mean that there's plenty of

30:31 there to feed everybody. Right? that's where the nutrients nutrients come

30:35 Right? So that's what kind of the favorable signal. So yes we

30:41 plenty nutrients more soda coming signal builds now we've reached that threshold and we

30:47 we can then go into this right . Right. Because that's the extra

30:53 soccer information that is induced. Gene expression of this material that only occurs

31:01 they've reached that threshold level. And so this quorum sensing,

31:08 That's the dependent themselves underground for and government you have people present to have

31:19 vote. Right? We have enough present, then you can you can

31:24 for bios, so to speak. . Right. So and and this

31:31 this quorum sensing phenomenon is not exclusive bio. So you see it in

31:38 phenomena. We'll see it later again uh transformation, transformation is the uptake

31:45 DNA from environment. There's actually some have it forms sensing mechanism for

31:50 Okay, so all it means is it's a sell it depends on cell

31:55 . Is there enough cells present? there is then enough chemical signals are

32:00 made and then that can trigger whatever process is in this case bilateral

32:06 Okay. Question. Okay. Um questions? Yeah. Yes.

32:21 What we're seeing is in how how do it? But what what differs

32:29 the chemicals signals that are used can more specific to that species?

32:36 So that's where you'll see differences. one species may use um a chemical

32:43 X. Another one using y. where the variations occur among the species

32:47 the overall process more or less occurred this. The number. I'm

32:55 It was a self density. What No I think that the cell density

33:03 can be achieved is strictly going to due to the nutrients available to

33:08 And of course the metabolism the cells . And so if you have the

33:13 all things being equal species X. a species why they're and they're being

33:17 their optimal nutrients then you know it's based on that that species intrinsic growth

33:22 which can vary but bottom line they're informed lots of cells. Yeah.

33:28 yeah, densities can vary depending on and these kind of things. Yeah

33:33 answer the question. Okay. Yes. So the bias information is

33:43 by a federal species. Okay now it's got in this in this um

33:53 like this and it gets big. ? Of course things are gonna get

33:57 in it. Right? So you get other species because they're there in

34:01 environment along with them that can get to the thing to be part of

34:05 . Will those actually be able to themselves and grow? I doubt it

34:11 they're gonna be so many of these formers there that the competition thing maybe

34:17 minimize their effect. But yeah it's species specific thing because they're in the

34:23 with other bugs. Those can get up in the mix as well.

34:29 other. Yeah. So so again biofilm Now we're growing up in in

34:37 uh all dimensions basically. Right. so this is where you can get

34:43 micro environments because you have cells on outside. Right? And you have

34:48 money interior. Okay. And so be differences in oxygen levels outside

34:55 Um Could be differences in how many the cells are getting outside versus more

35:01 the surface versus inside. Right. it can create kind of differences in

35:05 of metabolic differences. Can create growth . Maybe those out here grow faster

35:12 in here a little bit deprived. they don't grow as much and it's

35:17 medically important biofilm that can create differences perhaps antibiotic resistance. And so that

35:25 why these things can be a problem if they're of that type.

35:30 But you can what can happen is can create holes, holes can be

35:35 in these towers and that can kind help to circulate nutrients so that everybody

35:43 kind of get more now more to pie and get more nutrients and grow

35:47 can happen. Okay. But even that you still you still have some

35:51 of differences within the iphone itself when get so big. Okay. Um

35:59 . Any questions. All right. All right. Uh So this is

36:08 example of and those pores really example their level of um how long they

36:18 be viable. So they these are in fossils. Okay. Uh you

36:28 see that 2050 million years old. million years old. And the sports

36:35 are actually right here. That's a where it is. So this is

36:40 free and those four, that's And those four so they are quite

36:50 . Okay. And they've been able be revived. Right? Just like

36:55 seed that kind of seed, dirt water germination. Same thing can happen

37:01 an endospore. Okay, so the , the term spore itself um There's

37:08 kinds of dormant forms that microbes can into. Okay. There simply just

37:20 themselves which can be we all had fungal spores. Those are the type

37:25 reproductive structure. Right? Um bacteria protozoa can form spores. The system

37:36 another type of dormant form. Um the end those form when we put

37:42 we put these four letters in Okay. That is a next level

37:52 of resistance. Okay. Yes, and spores do have a certain level

37:58 resistance but nothing like an index for . Okay, so it's this too

38:06 a is a gene coded process takes genes to do this occurs over 8

38:12 12 hour, 12 hour process to this. And there's only two groups

38:20 in the microbial world or in the period. Use right clustering.

38:28 the sewers for the most part they're both soil organisms. Both soil

38:34 But for the most part relatively Okay, anthrax is gonna be the

38:40 that's the human pathogens. Uh and are a couple of others but most

38:43 them are fairly the soil organisms. harm. Australia is one that

38:53 Tetanus boxes. Um That's clostridium among . You see a couple of examples

38:59 gas gangrene is also caused by clostridium toxin basically causes tissue death. So

39:06 can see how the yes that's an of somebody's just basically turning black.

39:11 tissues gotten completely necrotic toxin killing the . Um regardless. Um in both

39:19 so metabolically speaking. Right. Very aerobic versus we'll talk about that.

39:29 first part of Chapter five will cover is relates to behavior the presence of

39:36 and how cells react to that. so um so the process of forming

39:43 those, right. Um it's a of differentiation into a different form.

39:51 . It's basically proceeded by let's just through the steps here. Okay so

39:56 a compartmentalization that occurs. You have sell. And then in the process

40:01 form two compartments and then one compartment the other. Right? And then

40:07 sets the stage reforming the spore. . And so we can look at

40:12 we look at those four forming Okay we see different types.

40:18 So vegetative growth basically a vegetative cell really just a normal. Okay you

40:26 consider yourself sitting here in your chair your the vegetative form of,

40:32 You're doing your thing. Right? So you better say that. So

40:38 what it does. Right? Uh if you look at a get a

40:44 of in those performing species. Look microscope these different forms. Okay,

40:54 self. So we're in those forests in silence. What's going through the

41:00 ? And one word done to Now you can gauge kind of the

41:08 of the culture if you will By all the proportions of these three,

41:15 was 99% educated. That's likely just healthy growing culture. If it's like

41:23 vegetative cells and vegetative cells with the board. And it's really going through

41:29 process. Okay? So it's And you can kind of gauge just

41:34 looking at these three different forms. . So you can also I don't

41:41 just put this in for for your information. It's not something I'm gonna

41:45 you on. But the the form can also like with the you can

41:52 like one single women or room where can be all around the cell.

41:57 with sports, they can be formed different ways and it's species specific.

42:02 way they form. Right? So types um some types will form uh

42:12 and a sport right in the middle the cell, right? Others forming

42:16 one end. Okay. Or near end. Or they can they can

42:22 inside the cell. Right? And these are species specific. Look up

42:27 the will say you know it's a it's a uh endospore forming cell.

42:34 Sport is I think uh central. and not swollen. So central not

42:42 . And that identifies it as is our species? So you can you

42:47 use that as a as a way I. D. As part of

42:51 identification. Okay so uh let's Alright so the process. So we

42:58 with replication of D. N. . Okay. And so it kind

43:04 forms a facility that they call it cell. Uh So we have two

43:11 of course then we can begin to the printing in here. But we're

43:16 to form two compartments. Okay so have a four score and we have

43:22 mother cell. Right so the four is going to form the sport.

43:28 . The end of the war will originate from that Four sport. Okay

43:34 what's the mother cell doing? Well is communicating. It's sending out basically

43:42 factors to the force board. And factors are initiating the expression of different

43:50 . Okay so kind of that's why call it the mother. So it's

43:54 of directing what's going on in this of the process? Okay so uh

44:02 the membrane of the cell. Mother somewhat engulf that for sport. Okay

44:11 so what happens is this D. . A. Of the mother cell

44:16 away integrates? Okay and so now have a double membrane around that fourth

44:22 . Okay um and then gets emphasized the middle of that. Okay and

44:30 actually call this this is what they , once the peptic like cannon gets

44:35 there, they called the cortex. then there's a very thick layer then

44:42 add in these chemicals calcium. And what's called die pick Olynyk acid

44:50 P. A. For short. and so that among other things buying

44:57 the D. N. A. um kinda helps protect the DNA quality

45:02 that in those four ft um It helps to reinforce that cortex also

45:12 the process. Um Water is drawn of this structure here of this structure

45:21 . This force for that now has cortex around it. Okay that water

45:27 drawn out. Right So that helps in its stability and what you might

45:35 long term storage. Okay. Water used to sell is heated,

45:42 So the damaging effects of temperature high come from the high water content in

45:47 living cell, 70% water Water begins boil and that affects the proteins in

45:52 cell, ultimately killing yourself. If remove a lot of the water then

45:57 you don't have that damaging effect as . Right? So in those sports

46:02 has like 20% water compared to 70% the living. So okay so that

46:09 in terms of its ability to remain for so long and still be

46:16 Okay so um and so then uh don't care about exasperated and it's kind

46:22 a transient kind of a structure. basically we're gonna form a the mature

46:27 coat once we've added all these chemicals it. And then so throughout this

46:34 here I'd say from probably here through hmm. About here. Okay those

46:50 Hear those 1 2. Let's call a day. B. C.

47:00 pen C. And D. Okay I'm sorry. That's when you see

47:07 cell microscope has I'll come right Has this appearance here? But they

47:17 inside of the cell. So it's through these A. B.

47:23 D. Stages these right here that that that that's what looking like a

47:30 . Okay then finally the free spore released. Right? So no no

47:39 cell part. And as you saw can last for millions of years.

47:49 problem to find these things like Egyptian right from like 20,000 years ago uh

47:56 elsewhere and have been able to revive . Right? So germination is the

48:01 where they go from endospore to a cell. That's called germination.

48:08 So that can happen under. So induces this um all kinds of

48:13 Deprivation of nutrients right? When it gets to one of these bacillus gets

48:21 stationary phase and in the stationary phase typically when they start forming in the

48:26 . Okay um New lack of nutrients any kind of other environmental stress radiation

48:35 temp cold temp um ph right? treatment. Any kind of stress can

48:45 induce endospore formation? Okay. And course what reverses it is favorable conditions

48:53 be and the sport can then germinate grow back into vegetative cells again.

48:58 . But the endless wars super Right? That's why we rely on

49:04 get rid of it. How to conditions create steam. They use the

49:09 at high pressure, right? So get temps of like above 120

49:15 Um And um that combined with the heat of steam, can penetrate the

49:22 of four and kill it. Um anything else? Any questions english

49:33 Um Yeah, simply just if you're a free sports state here you can

49:45 back to a vegetative state that under conditions, right? That's germination.

49:52 think of a seed which is basically dormant thing. So in the ground

49:57 water it grows or germinates same So the endless poor going from its

50:04 state into a living breathing, vegetative that processes germination. Yeah. I

50:11 he said that. How would you me? That's true that you have

50:18 do two things. One is you to cut out the dead tissue and

50:22 you have to wash it out or . But then of course you have

50:25 give antibiotics and about the freedom ultimately gonna kill it if it's not or

50:30 have to cut a limb. It too bad. So, uh,

50:35 other questions? Okay. Um, branding is not something very common these

50:41 . It used to be back, know, in wars and things when

50:44 have like wounds and you didn't know about exceptions and what not a lot

50:48 times. That's where you would see . Um Okay, so transfer chapter

50:55 . Chapter five basically cover a couple things. First part of this is

51:00 call it zero tolerance. So, you have a bacteria or archaea and

51:08 know, and you often Okay, can be different responses. Okay.

51:15 of course it relates to, in large part relates to what's the

51:20 themselves. Okay. And so there be five different responses. Okay.

51:26 on the cell type. But kind , overall this chapter we don't go

51:32 the first part's really kind of First part of the first half of

51:35 chapter is about title right here. ph osmolarity affect growth. We're focusing

51:46 oxygen. Okay. But uh just at very briefly at temperature ph and

51:53 . Okay. Every microbe has every has its optimal in terms of temperature

52:03 ph which they grow best at. ? Um it all relates to the

52:11 thing, but temperature osmolarity, ph what the optimal conditions are. What

52:18 their proteins happy. Okay. For temperature ph or popularity, Those three

52:27 influence protein folding and shape, treasury of protein. Right? So that

52:33 strands are together in a certain Right? And that shape is due

52:38 hydrophobic interactions, Charge interactions. Um bonds. The sulfide bonds.

52:46 So, ph a sub optimal ph or hospitality can affect those interactions.

52:56 . And make a protein not work . Right. So, you

53:01 the optimal temperature ph osmolarity or what for whatever it is for that

53:06 It's it's that those conditions because that's keeps the protein test pretty happy.

53:13 , they'll like that. Okay, in a nutshell, kind of what

53:17 about. So, you have most are in the middle right temperature of

53:28 book gives a slightly different range, say 18 17 18 C too?

53:36 . Okay, everybody's kind of in range for don't worry about writing this

53:41 . Music files. Everything probably Most things operate at that around ph

53:47 . Right. New travel files. . Um osmolarity. Um typically about

53:56 can't think of the average value, um in marine environments. 3%.

54:02 freshwater environments or traditional elements. Maybe or less .91%. But then

54:10 this is where the majority of Is that these values for these?

54:16 you have extremes, right temperature. , right. You can have hydrothermal

54:21 . Right. Key. And we about those guys, you can have

54:24 profiles which are like very cold Right, extremes of ph Right.

54:31 , the point is, these parameters about taking proteins happy and most living

54:36 are within what we call moderate ranges these values. Right. But you're

54:42 have some few months on either Right. Not the majority. But

54:46 can find them in these certain environments they have adaptations that enable their proteins

54:52 be happy at these fringes. so bottom line by keeping the proteins

55:00 . Right? And so we are to focus on um 02.

55:07 and so this and then the second of this, um the rest of

55:15 , I should say it's about Alright, so there's gonna be

55:20 There's gonna be some uh examples of you can control physical parameters. Chemical

55:29 . Um that's kind of what that's . Okay, so let's start with

55:33 tolerance. And so if one is living on this planet, okay,

55:44 are uh supposed to air. Usually. Okay, uh were

55:54 Obviously, we need we need Okay. But whether you use oxygen

56:01 if you do not, you will be affected by it. Okay,

56:09 auction is very reactive. Okay, it likes to grab electrons for

56:15 Okay, It's oxidizing agent. very strong. And so we of

56:23 , like many others have an aerobic . Right? We'll focus on that

56:27 the next unit. But for you know, we have uh we

56:32 down an energy source and in the we use those electrons and ultimately hand

56:40 off the oxygen. Okay. So auction itself can react upside reactions.

56:50 ? The action itself can interact with . Okay. And do their kind

56:57 oxidizing thing. Okay. And so can form what are called reactive species

57:03 oxide hydroxide radical hydrogen peroxide. Okay these damage molecules and some particularly Vienna

57:12 proteins is um in our own might have heard of foods that are

57:20 . Right? Blueberries and things like . Okay because they help to to

57:27 the detrimental effects of of this Help help to eating this kind of

57:36 support supposedly help to counteract the damaging and allow us to you know,

57:44 damage our cells and tissues. Believed to be a contributing factor to

57:49 and and death. Okay. So any case um so if the face

57:57 living in In air whether it uses or not, it's gonna have to

58:02 with this issue. Okay. Or will die. Okay. So you

58:07 to have protective enzymes that will counteract effects of these reactive species.

58:14 Because these are gonna form again regardless whether you use option. Right.

58:23 these enzymes do what? Well they the effects of these species. So

58:30 oxide radical S. O. Which is shorthand for super oxide Disney

58:36 . Okay uh and cattle A's and . So super oxide is mutates forms

58:46 peroxide which is still reactive not as as super oxide radicals but we have

58:52 A's and proximity is to neutralize the of hydro peroxide. So of course

58:58 end products here water are and oxygen of course. Sorry. The water

59:07 completely benign obviously. Right, so form water now you neutralize the effects

59:11 these species. Okay, so um again, microbe that's living in this

59:23 world. If it uses auction, doesn't use oxygen, it's got to

59:30 with this other issues production of these oxide radicals because they will form in

59:40 and if they can't deal with it they will die. Because if they

59:44 have their protective enzyme, these reactive will damage their proteins and nucleic acids

59:51 they will die. Okay, so have a couple of choices here if

59:57 the micro have have the protective enzymes protect yourself or get away from there

60:06 together, don't even be in his . That will avoid a problem as

60:12 . Okay, so think about Let's um look at how in the

60:20 and kind of find out an organism's to oxygen. Okay, so you

60:27 a medium like this that basically when have a medium where you can get

60:33 oxygen gradient. Okay. And so medium called fluid like light will do

60:40 . Okay. And so um it's gel like medium. It has chemicals

60:47 help remove oxygen when you make it then uh put it on a

60:55 The gas boils out of it then it gets to room temp comes back

61:00 , if you got affected air will seep in. But chemicals in the

61:07 will kind of get rid of Not all of it because you've been

61:11 radiant. Okay. And so you here from high too low at the

61:20 . Okay, so there's your un reading, watching high to low or

61:28 at the bottom. Okay. The is a dye, the red color

61:32 where oxygen is present. You can right at the top progressively less as

61:37 go down to. Okay and so you do is you take a wire

61:44 and whatever you're going to test and inoculate in a way that you're what's

61:51 stab? Just stabbing it Viagra going the length of Viagra. And you're

61:56 the loop back out. Right? you're basically seeding the cells you have

62:03 um that are laid all the way the length for the medium.

62:08 so now what you're asking is what which sells? And what region are

62:15 in what region of the growth is be only those that are down

62:22 Only those that are up here or in the middle. And where do

62:29 see the growth? And what part to challenge you their response to

62:34 Okay, so again, we're asking question right, looking for a pattern

62:42 growth growth to the bottom of the to the top or throughout the whole

62:48 . Okay, so all those will you something about it. Okay,

62:53 here's a question. Okay, so just gone through that. So we

62:59 five strange A. Through E grown this same kind of medium.

63:06 Um Based on the results from which strain A. B.

63:16 D. E. Is not lacking protective enzymes but has reduced levels of

63:25 enzymes. Or maybe it's missing one two of these enzymes. Okay.

63:30 not completely lacking has some. Right What kind of response are they gonna

63:39 ? Okay, this is Okay. the timer going. Alright, Count

64:29 7, 6, 32 and Okay. Um Mhm. If you

64:51 uh e Okay, so let's look this. Well first let's let's go

64:58 the different types then we'll come back this. Okay, so um the

65:06 what we call a micro verify. , so let's look at the different

65:12 . Okay, so uh let's break down this way. Arab and Arab

65:19 . So Faculty Ativan Arabs can use not use oxygen. Okay. E

65:24 is a faculty of an arab. So the Arabs. And so they

65:31 distinguished by the fact that they use , oxygen using metabolism. Right.

65:36 that we are right, but maybe types. Okay, my profiles are

65:43 . Are they're lacking. Like the said they're lacking maybe one or one

65:48 two of the enzymes protective enzymes where have lower levels of protective enzymes.

65:53 , So that means they don't have full full protection you need against atmospheric

65:58 of co two, which is 20% or take. Right? So they

66:04 like 10% 1% option. Too much them because they don't have enough protection

66:10 it. Right? That's why they like in this level here, not

66:16 the top, but somewhere in the . Okay. Um the and a

66:23 . They do not use oxygen. . At all. They typically aren't

66:32 . They ferment or and or what call anaerobic respiration. They breathe with

66:39 other than oxygen. Okay. But can have two types. Okay,

66:46 as the name of the obligate, obligate must be That's that's the ones

66:53 that strategy is get me away from altogether from air. Getting away from

66:57 . Okay? Get me away from don't have any protective enzymes.

67:02 So I must be away from it together. Right. That's why they

67:07 the very bottom. They don't have protection. So they're gonna be right

67:13 there. Right. No protection. give you away from it all time

67:17 . Okay. You can be an tolerant. And that's the one that

67:22 confuses most people. Okay, the tolerance, so that the name

67:27 It doesn't use option but it can it actually has a full problem has

67:35 protective enzymes. Okay. And so what enables it to live in an

67:42 environment Because it has the max protection it. Okay. Even though it

67:48 use it enables it to live in 02 world. Okay. The faculty

67:55 antelope can use it or not use . Right? E coli everything is

68:00 type. Um So the they have whole protection options, not world,

68:12 environment. And so when we look these patterns. Right? So here's

68:17 obligate strict arrow. I think the this one one that's probably the most

68:27 . Are these two? Alright, call this. That's A. This

68:32 B. And see right there. anaerobic faculty, those the ones that

68:38 confusing. So what you have to , you have to differentiate growth at

68:44 top. Right? So you can here there's more growth at the top

68:50 your faculties because oxygen using oxygen gives a lot more energy. Right?

68:58 energy equates to more sells more Okay, so in fact, in

69:03 robe will have more growth at the compared to the zero tolerance and zero

69:09 , zero tolerance arrow is not getting advantage by being at the level of

69:13 medium where the most option because it use it. Okay, so it's

69:17 or less kind of distributed evening throughout tube in terms of cell density.

69:24 The fact of the matter opens through , but they have a little more

69:26 on the top because Oh, to , more energy, more brooks.

69:34 . Any questions makes sense. Are . No, that's okay. That

69:44 sense. Right. Okay. So and we'll learn about the

69:49 Uh and you have to So let's at um let's look at a man

69:56 come. I'll show that. Show question next time. Um let's so

70:02 I'm going to transition into a little about ways to control growth.

70:07 So, we've all seen our favorite our favorite in this case. Disaffected

70:12 right, Life's all very common. all have a label like this.

70:17 because all these kind of antimicrobial agents products rather are tested against, you

70:25 , uh gram positive gram negative against against viruses. Okay. And of

70:31 it's written on the label. What effective against. Okay. And various

70:36 like uh reading the label. It kills foods and viruses. It kills

70:41 and flu viruses, uh disinfect blah blah sanitize, etcetera. So,

70:46 focusing on here is the value. always has some big numbers, kills

70:52 99.9 99.99. What have you? , so people often get confused?

70:59 , well this is sterilizing the I'm using. Okay, just to

71:04 you, you know what actually is in terms of numbers. If we

71:08 a surface, you know, and test these things on like a a

71:12 square inch surface, a little Then they'll swab the template and get

71:17 cell count. Then they'll take the template and then get an area and

71:21 treat it with this effect and then how many were killed. Right?

71:25 let's say one million cells. We apply our Lysol and we get

71:32 many cells were killed. It was 99.9 being killed. They were killing

71:38 cells. Right? We still have cells left Bible cells. So we're

71:42 going to zero obviously. Okay. what we call logs of death,

71:49 the parameter used in kind of quantifying well one of these agents is

71:55 The manufacturer looks at how many logs death are we getting? And what

72:00 frame are we getting those logs? . Because obviously quicker is better.

72:05 , and so um so we'll talk next time, but the logs of

72:11 is a round that we're gonna be at. Okay, remember death is

72:16 rapid but just as gross. Can very rapid. Okay, so we

72:20 to maximize that. We're trying to their growth. Okay, So here's

72:25 question about terms. Okay. And you know that my pet peeve is

72:30 of sterilization. So let's see if let's see uh remember that.

72:37 so remember sterilization? Not not in context of can't have kids. Right

72:45 of killing stuff. Okay, it's okay. It's okay to

73:18 You know, I'm going to say okay to pick this one. You

73:24 what I mean, hint, It all makes sense. Then please

73:32 this. Yes, you did. . None of the above.

73:39 None of the above. Okay. not eat. Alright. Here's our

73:48 told you at the beginning I'll come to that next time. Here's our

73:52 bowl question. Mhm. I realized don't really watch this football. Which

74:00 why A and E. For Sure. What's up? What's

74:20 My home? Is that a Yes, I have a This is

74:40 no correct answer here. Let's see we got. Yes. All

74:49 Oh, okay. Okay. That's majority folks. See you next

75:00 Okay. You got it right. matter what? Oh, this species

75:46 were created by now. Flying Yeah. The day is if I

76:09 finish, so we're gonna do that monday. I went through everything for

76:16 unit. Typically, they're not usually because I'm pretty good about being getting

76:24 done, but that's what I Is that this one uses oxygen

76:40 This one thing. Yeah, I say so. I but I would

76:58 when I tell people there's two there's in the honors college, you

77:04 have to be any harm in but it's just what the local it

77:06 called. The office of regular So they're like, they're like really

77:13 college on the second floor. There's big sign you can't miss it.

77:16 they will have a list of labs have opens to though is go to

77:23 go to the biochemistry biology website. can just access to sm biology department

77:35 we're just gonna research faculty and see they're doing if you want to do

77:40 . I'd say the two professors are Roger who is a biochemistry and he

77:48 takes on undergraduate students. The other is dr he does research. DR

77:57 a variety of things but he's definitely method biologist. Um And I'm not

78:05 sure that others. There's actually you expand your search to even the chemistry

78:12 working microbiology work in chemistry. So might check my check there as

78:19 Even even pharmacology. Okay pharmacology is got

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