Transcribed by Amit Amin

10/31/2014

[Infectious Diseases] [1&2] [Microbial Pathogenesis] by [Dr. Boylan]

[1] [Microbial Pathogenesis]
[Dr. Boylan] We have a course in infectious diseases and on your syllabus you
should be able to find the lecture schedule and the info on the faculty teaching in
this course, as well as their email addresses and where they can be found in the
school. There is also information on the examinations. There are 3 examinations, the
date you should have also in your block schedule I believe. One is in mid-December
before the break, one in January and one in February. Each worth about a third of
your grade. There will be bonus points for attendance at conference. If you get to 4
out of the 5 conferences you get at least a point added to your overall average.
Maybe more, well see how the class does on the exam to let me know whether one
point is needed. The final conference (week before exam) on immerging infections
and bioterrorism will not be on the final exam but well have a quiz thats going to be
worth up to one or two, maybe even three bonus points. Last year I did it and had a
competition b/w the sections. Kind of like the Olympics b/w the sessions. The best
grade had 2 points, etc. It worked out pretty well actually. I had a video I showed
that you had access to look over the night before. Thats about it really. IF you have
any questions, everything is there about makeups, remedial. Ill get this posted too.
The syllabus will have an outline for every lecture. It might help you prepare for the
lectures given by the different faculty. Microbial Pathogenesis. We will talk in this
course about infectious diseases caused by our friends, the bacteria, microbes,
viruses, fungi, and the protozoans or parasites as they are called. We had the course
on microbiology in the summer. You are introduced to these topics more or less and
you know the properties of these microbes. Bacteria, the cell walls, flagella, Pilli,
capsule, etc. The viruses, their structure of how they replicate in our cell and the
fungi as well is one of the lectures given. WE will get back to those and talk about
some of the damages they can do us and infectious diseases. If you want any
information on Ebola or any other microbe, the best source of all is to go to cdc.gov.
That site gives the most updated information about any of these infectious diseases
were going to talk about. What they are, the symptoms, the signs, how they are
treated, incubation period, everything like that. On cdc.gov. In addition to that CDC
comes out w/ a weekly report called the morbidity and mortality report. Morbidity
means people getting an infectious disease/ ill, and mortality is when people die.
Every week from about 30-40 different infectious disease a health care professional
must report that to them. This is so that they can keep tract of any outbreaks or
epidemics w/ infections. They know, week-by-week they study how much hepatitis
B is occurring in NY, Cali, Northwest, Northeast, etc. They can see any increases in
incidence of an infection and they go whats the problem there? Why is there any
outbreak or an epidemic of infection? Why are there high numbers? Talk about
dumbing down the language. Today, they were talking about these people who do
this, go from the CDC to a site of outbreak or infection are called epidemiologist.
Epidemic occurs or an outbreak. Now they are being referred to as disease doctors.
Thats what they are I guess. I still like the word. Whats wrong w/ epidemiologist?
Its a long word. Cormo? Governor of NJ whose name I dont want to pronounce, he
was a disease doctor. Trying to tract down/ handle Ebola patients. CDC.
1

Transcribed by Amit Amin

10/31/2014

[2] [Infectious Disease Deaths]

[Dr. Boylan] Infectious disease worldwide. This as you can see here, in the 1960s in
this country the Surgeon General of the U.S. said we have infectious disease problem
pretty much under control. In a few years we probably wont have many infectious
diseases. They will be so minimal; there wont be a concern about that. He said that
b/c we have antibiotics to help treat infections; we have good vaccines to prevent
many of the most dangerous infections that occur in childhood. He was of course
completely wrong. We still have infections with us. We are in many ways
responsible to the increase of bacterial resistance in bacteria to infections. Also,
immerging infections that have arisen past 30-40 years so have long let it go if we
can ever eradicate infectious diseases. I dont think we ever will. If you see here this
is worldwide. This is a couple years ago, 2010. TB, I dont want you to memorize
these numbers, but you can see the problems we work with. We have people dying.
People die worldwide; the percentage of deaths attributable to infectious diseases is
about 15%. 15% of the worldwide populations still die every year from different
types of infections. Many of the common ones are shown here. HIV/ AIDS, diarrheal
diseases, gastroenteritis. 1.8 million mainly children. We think if we get
gastroenteritis, ok, its a mild problem for a few days but in some countries where
they have these infections of viruses and bacteria this diarrhea leads to a great loss
of bodily fluids from my body, and if they dont have a source of replenishment of
these fluids (water, salt solutions), the body just dehydrates and they lose just not
the water in their body but the salts and the electrolytes that are in ourselves so
they die. Its a very serious problem. There are other diseases like diphtheria,
whooping cough, others that many years ago would kill many (hundreds of
thousands of children) in this country and worldwide. We have better vaccines now
for many of them. Malaria, tropical diseases, respiratory infections, Hep B/C,
shigellosis which is a dysentery bacterial, also great loss of fluids in the body,
dehydration and death. They are still with us these infectious diseases. Theres
concern about them of course.
[3] [Colonization]
[Dr. Boylan] When are we first exposed to microbes? We will talk mainly about
bacteria but some viruses will be also included as well as the fungi to a lesser extent,
parasites. So, were focusing on bacteria in these next couple hours we talk
infectious diseases. Welcome to the world, when we are in our mothers womb if
you can think back that far, remember when you were a little fetus in your mothers
womb you were sterile. You didnt have any bacteria at all, any viruses. Nothing.
Sterile. Thats good for us because we dont want to get infections when were in
utero. There are some bacteria that can be transmitted transplacental from mothers
to child/ fetus but ordinarily sterile existence, no bacteria when youre in the womb.
Newborns, contacts microbes during passage through the birth canal and from a
mothers skin. There are different ways a newborn is exposed to microbes. Feeding
materials, instruments used by health care professionals, contact w/ other people,
nursing as well. Then we get what we call the normal microbiota. Microbiota. Living
things that live on us and in us. Thats of course also known as the microbiome.
Micro, all the living creatures (microbes) that live on us and in us, microbiome. This

Transcribed by Amit Amin

10/31/2014

microbial exposure is renewed over and over again. So lets talk a little more about
this phenomenon.
[4] [Colonization of Newborns]
[Dr. Boylan] They have done studies to see if there is a difference in the microbiota
of children who are born vaginally (normal) or C- section to see if they indeed
harbor different microbes in part of their microbiota. Heres the newborn on the left,
here you see vaginal birth and on the right is C-section birth and vaginal exposure to
microbes especially from the birth canal of the mother and the skin of the mother
and if it were a C-section, more bacteria accumulate on the body from
environmental microbes (other people in the nursery, other ways we just saw on
the previous slide). The point here is that even if there are differences in deliver,
either C section or vaginal, you can see that this tells you the differences in the
microbial population in a newborn from Day 1- Day 200. So yes, depending on the
nature of the birth itself there can be differences in the microbiota of the newborn.
This continued for a while. Nursing can also effect the microbiota of the infant.
Whether youre breast-feeding or giving formula, or the mother has high maternal
(some word I cant decipher) bacteria, environmental, etc. Finally after about a year
the microbiome of a 1 year old is the same as it will be when they are adults. What
you have now is what you had about 1 year. There are indeed fluctuations in the
percentages of the microbes perhaps but more or less the basic components are the
same but depending on internal and external sources, internal- antibiotics/ meat
free diets, tropics environment, other infections, these things can for a time affect
the microbiota you have but it always goes back to about the same levels and the
same microbes and they differed among us. We differ very much in our microbiota.
For each individual we can use this as a fingerprint for each individual. What
microbiota they have. We differ very little in our genomes. Maybe .1% or something
like that but in our microbes we differ quite a bit more than that so genomes wont
tell us as readily as the microbiota. In the future, that could be a great deal to us in
determining what treatments we want to have for some infections or other things
we will talk about later as well. So the human microbiome project is going on now,
as we look at the different types of microbes found on people.
[5] [A few words]
[Dr. Boylan] Commensals, ok, the human body, I think Dr. (cant decipher) likes to
use this methodology/ terminology is like a cylinder the digestive tract from the oral
cavity to our inner opening, a cylinder w/ a central core, skin, eyes, nose, oral cavity,
intestinal tract, urogenital tract, they all have bacteria (commensals). Ordinary
commensals are referred to bacteria that can derive some benefit from living on us
and we dont have much of a benefit but well see that indeed we do have benefits
from having this microbiota on us and in us. Beneficial roles are shown here for
example. By having these bacteria coating our surfaces of our skin and our digestive
tract we are protected from infection. We are probably exposed to a lot of microbes
on a daily basis. Some of them maybe be potential pathogens that we inhale, we
digest, we drink, but if they cant find a foothold in the body to adhere, they have to
stick to something, viruses, bacteria, fungi, they wont cause any infection at all, if
they cant adhere to our tissue so just patch right on through and be excreted or
3

Transcribed by Amit Amin

10/31/2014

coughed up. The pathogens comes in our body, the potential pathogen can cause
disease/ infection. They have to compete w/ our normal microbiota to get that
foothold, to adhere so our microbes protect us by coating surfaces and using up the
nutrients there that pathogens might want to use but they cant derive them b/c
there are bacteria there to protect us from that happening. Our normal microbiota
also stimulates our immune system. Youve had lectures on immunology, I know
about our natural and acquired immunity but its ready to go, naturally meaning
there from birth, innate immunity, from birth. Where acquired immunity, T cells/
antibodies come later on but the acquired immunity is primed to get ready to fight
off some infectious agent by just being exposed to our normal microbiota saying ok
we are aware that these bacteria are here in our body, well keep them under
control and not let them grow extensively and that doesnt do damage to us. But
were ready we know what to do now in case some other bacteria/ microbes come
in. Were primed to respond rapidly w/ our acquired immune system to fight them
off. This just generates stimulates waiting to happen wed be overwhelmed by some
new infectious agent, if our body wasnt primed to respond by our normal
microbiota. Nutrients also, provided by some commensals, microbiota, some
bacteria thrive in our gut can produce vitamins B/K that our cells in our gut can use.
We do have benefits from microbiota. Internal organs usually sterile, the liver, the
blood stream, urine in the bladder is sterile, when it leaves the body well see its not
sterile. When you collect urine it wont be sterile. When it passes through the
urethra it picks up bacteria there but in the bladder its sterile. Heart, lungs, liver are
ordinarily sterile. Only our skin, oral cavity, and in the picture coming up other sties
that might be colonized w/ bacteria.
[6] [A sampling]
[Dr. Boylan] Just to give you an example so you dont have to memorize these
things. Samplings of the indigenous microbiota of the human body. Indigenous,
whats there on us/ in us all the time? These are the bacteria we all carry w/ us.
Different ones in different proportions perhaps esp. in the gut. Upper respiratory
tract. The nose, nasopharyngeal area, oropharynx, bacteria that are there. The skin
has 120 different species of bacteria and fungi. Female reproductive tract, vagina,
lactobacilli, staph and other bacteria as well as Candida. Candida as you know is a
yeast, fungus that causes yeast infections in females routinely. Oral cavity, up to 500
different species of bacteria. Not all of the same time, but we probably have a couple
hundred ourselves in our oral cavity, on our teeth, in our gingival pockets, in our
buccal mucosa as well. Urinary tract, urethra. As the urine leave the body, they will
pick up lactobacilli, and other bacteria from the female/ male urethra. Intestines
have over 1000 species. Small bacteria, bacteriodes, lactobacillus, strep, large dense
microbial population. A lot of bacteria. Up to a 1000 species in the gut.
[7] [Some of our normal.]
[Dr. Boylan] Some of our normal resident microbiota. Eye is similar to the skin. The
skin has a lot of; the skin as far of bacteria is concerned is kind of salty and has a lot
of lipids in the skin/ tissue. Some bacteria like a salty environment, maybe a low pH
as well, and some lipids. S. Aureus and the members of the genus staphylococcus
4

Transcribed by Amit Amin

10/31/2014

they grow very well on our skin. The most predominant on our skin is S.
epidermidus. Epithelium, epidermal tissue but S. aureus can also be found on the
skin of the majority of us. S. Aureus were going to talk about that next week can
cause a lot of different infections from mild to very severe and it is found on the skin
of people and also in the nose and in the anterior nares in the front of the nose. This
is where S. aureus likes to colonize our tissue. Skin, its a bit cooler on our skin than
inside the body and the tip of the nose/ anterior nares is a bit cooler than the back of
the nose and the rest of the body. Its a 2 degrees different (35 C rather than 37 C)
but S. aureus prefers that 2 degree drop in temperature to thrive. Thats why it likes
the anterior nares. The S. aureus in healthy populations, about 30% of our normal
healthy population of humans are carries of S. aureus in their nose. The problem is
that health care workers that are constantly exposed to S. aureus in a hospital
environment and the patients w/ staph infections about 65% of health care workers
esp. those that work routinely in health care settings in hospitals harbor S. aureus in
their nasal passage, in their anterior nares, thats the greatest site for infections in
hospitals. This site, the bacteria, the S. aureus found in the nose of health care
workers. Your hand touches your nose, even when you wear gloves sometimes; you
touch the patient, the bedding, towels, doorknobs, and spread of S. aureus infections
in the hospital environment. Oral cavity, we have viridians streptococci. Does
anyone know what viridians stands for here? Greening. Exactly. Greening
streptococci. Streptococci come from those bacteria that come from the genus
streptococcus. Notice there is no italics b/c its referred to the group of the bacteria
in that genus (cocci) if its coccus it would be italized. The v. cocci are those that are
found in the oral cavity that are green. Why are they called greening strep? B/c we
grow these bacteria in blood agar plates, petri plates w/ 5% sheep blood that we
add. Plates that are nice and blood red b/c they have sheep blood in them. We streak
the bacteria on the plates. They form colonies after a day of incubation and these
colonies of streptococci you have partial hemolysis of the red blood cells in the
blood agar and they look kind of green. ? There is a zone of hemolysis of the blood in
the blood agar plates around these bacterial colonies. Its partially hemolyzed red
blood cells and theres a greening color behind the zone around the bacterial
colonies. Thats why they are referred to viridians streptococci. Viridians is not a
species, its the term used to describe these streptococci that turn the color of the
blood agar green in the zone around them when they form colonies. On the teeth
you have S. mutans, the main culprit in caries. Not everyone caries it, but most
people do. Plaque bacteria also, the plaque above the gum line is the supragingival
plaque and below the gum line is the subgingival plaque. These are the codlings of
bacteria in organic film that we have to floss everyday to get rid of, were never
going to get rid of all of them of course but the supraginigival plaque are the bacteria
that are mostly aerobes or facultative bacteria and the subgingival down below the
gum line, down there in the pockets, gingival crevice, gingival sulcus you have a lot
more anaerobic bacteria. So the types of bacteria found on the tooth whether the
sub-or supergingival they vary quite a bit. Once again the subgingival bacteria
meaning anaerobic bacteria and facultative. Are there any potential pathogens in the
nasopharyngeal area? Yes I mean S. aureus is there but a lot of bacteria that cause
very serious infection, like pneumonia and meningitis, are normal residents of
5

Transcribed by Amit Amin

10/31/2014

humans. Up 30-40% of some of the most deadly pathogens. They only effect
humans, some of these bacteria, they are only found in humans not in animals. We
carry them in our oral cavity, in our nose, and there are sites of transmission from
one person to another. Theyre there. They dont want to make themselves evident
all the time, they dont want to cause everyone to get sick or death b/c then they
would die out or not be able to continue causing damage to us. They are there but
they are sneaky these bacteria. Were going to reside here and be unnoticed. Not
going to be seen above the radar. Occasionally we will decide we are going to cause
an infection, a serious infection like pneumonia, meningitis, and some others. The
point being here, even some of our normal microbiota, that we harbor, about 30% of
us well talk about have some of these most dangerous bacteria that are ordinarily
non-pathogenic but since theyre carried only in humans, they must be the ones are
the source of the infections that occur later on. Well talk about why they change
from normal microbiota to pathogens; well talk about the individual bacteria, what
signals them, what triggers them to become pathogenic bacteria.
[8] [Intestinal Tract]
[Dr. Boylan] Intestinal tract, also, stomach. We use to think it was sterile since the
pH there is 3. We know that the pH of the stomach helps digest the food but indeed
there are bacteria, one very famous bacterium b/c the guy who worked w/ it won
the noble prize about 10 years ago. Anybody know what that bacterium is? It causes
ulcers. E. Pylori. Very interesting stories about that how its able to survive in the
low pH of the stomach. What does it do to protect itself? Theres kind of a shroud of
protection around it to survive in that environment. There are some bacteria there.
Very small number. Small intestine, up to 10g for fecal matter, large intestine as
well. Anaerobes. Those are the same numbers we find in dental plaque. The
concentration of bacteria per gram of dental plaque is the same as fecal matter. Its
very rich in bacteria, different ones but very high counts. In the gut weve got a lot of
bacteriodies and fusobacterium. These are two anaerobe genera of bacteria and
they provide 50% or more of the microbiota found in the gut. Ordinarily theyre not
dangerous. Theyre there. They prefer to live in the gut. They like that environment
but theyre not a source of too many infections unless they get out of the site where
they are ordinarily found. Any bacterium, if we hold it in check, if we keep it in the
site its supposed to be found, fine. If they get out of the site where they are
ordinarily found into some other sites via the blood stream, via some injury, thats
when any bacteria can potentially be pathogenic. But if we hold them in check, keep
them at the site of the body where they are ordinarily found, theres usually not
much of a problem. They an cause infection if they get to another site. How about E.
Coli. E. Coli is indeed in our gut. All of us carry it. Very small percentage of the total
gut microbiota. Only about 0.3% or less is E. Coli. Ordinarily, E. Coli is harmless to
us. Its one of the ones that provide some vitamins for us when it grows in the gut. It
protects the lining of the intestinal tract against the infections by other bacteria. We
eat we drink, get to the gut. Why isnt this E. Coli thats part of everybodys normal
microbiota cause diarrhea, outbreaks of gastroenteritis, septicemias, UTIs. Why do
E. Coli, if we all have it, does this? Doesnt the body keep them in check? The
problem is we are exposed to E. Coli from other people, sources, or contaminant
6

Transcribed by Amit Amin

10/31/2014

food or water and these are different strains of E. Coli that arent ordinarily found in
us. These are the ones that usually do damage. Were exposed to new E. Coli strands
that our body does not recognize. This E. Coli thats not supposed to be there in our
body so it responds. Another source, yea. Right. Matter of fact, Ebola. There are
probably a lot of infections that we have come from animals. They are species
jumping like the Ebola virus. Its reservoir is in fruit bats in Western Africa. In that
host, yes it replicates and its present but it doesnt kill or cause hemorrhaging in the
bat the way it does when it jumps from the bat to a human where our immune
system cant control it. I hope we get to Ebola later because learning about the
factors it produces, it causes terrible disease, hemorrhaging all over the body. Its
amazing but yea, many infections do come from (some word) also, animal to human
infections. Well talk about those as well. Matter of fact, I meant, would have had it if
it had been published on NYU Classes a whole list of terms used in infectious
diseases, the definitions of them. I think you should look that over as well. That will
probably help you. We will define most of those terms on that handout infectious
disease definitions but its good to have a source at the ready if you want to look
over those things. So were just getting an introduction on infectious diseases today.
[9] [Harmful effects]
[Dr. Boylan] Harmful effects of microbiota. Breach, once again, when they breach
the defenses we have. We have a cut in the skin, a lesion somewhere in the body,
another infection, our innate immunity trying to hold them back is breached. The
umbrella, our macrophages, our skin, interferon, things a part of our innate
immunity, if those things are breached, if they are destroyed or not produced, if the
cut in the skin, then these normal microbiota get to a new site underlying the
epithelium. Subcutaneous layers getting into the blood stream. Then it can be very
serious infections. Gets into the blood. Following antibiotic therapy. Whats the best
example of an infectious that might arise in a human following antibiotic therapy?
Which one? Yea, ok. What do antibiotics do? You use them, take them to treat a
bacterial infection right? Ok. Yea, the antibiotic works on those bacteria. Cuts down
their numbers we hope after their exposure to it. Either kills them or stops them
from growing but it doesnt just effect only that microbe causing the infection.
Antibiotics have an effect on our normal microbiota, maybe suppress them for a
little bit too, but what about other microbes that are resistant to that drug? Fungi,
yeast. Antibiotics you give to a person to treat a bacterial infection, the yeast are
resistant to those antibiotics. They are not killed by them, so they can flourish.
Youre going to see that a lot. Dr. Phelan is going to talk about that a lot too. Yeast
infections, thrush, or oral infections you suppress the normal microbiota w/
antibiotics, resistant microbes like fungi, candidiasis can emerge. So candida always
there but not causing any damage but when it sees its neighbors are disappearing, I
can use all those nutrients that bacteria were using to flourish and grow myself and
then grow into our tissue and cause oral infections. Of course, in
immunocomprimised, meaning that their immune system is not functioning well.
Its compromised in some way. HIV infections for example. People have a
compromised system b/c their T cells are lowered and once their T cells are
lowered, as they are destroyed and HIV progresses, we dont have the general alarm
7

Transcribed by Amit Amin

10/31/2014

of the immune system to help us respond to any infectious agent. Even really
innocuous can all of a sudden become pathogenic when our immune system
becomes depleted, begins to wane. Things like that, lead to the emergence of
infections. I call them endogenous infections, caused by our own commensal
microbiota bacteria when things like this happen. They can also cause deadly
infections.
[10] [Events that take place]
[Dr. Boylan] In general, we talk about infectious diseases, just about infection we
will talk about in this course follows the progression shown here. Makes sense of
course. Encounter, you have to encounter the agent, the parasite, and the microbial
cause of the disease. Either you get them at birth and they can remain quite
quiescent part of our normal commensal bacteria/ microbiota, the entry of the
agents, enters the host, spread from the site of entry whether its an abscess in the
skin thats going to be, it spreads from that site of subcutaneous underlying tissue,
causes infection in the gut doing more and more damage and spreads, travels
through tissue, via the blood stream as well if it gets in there. They have to multiply
in the host. Normally we are not infected with a large enough dose of microbe to
cause an infection right away. Ordinarily, the microbe has to grow, multiply,
replicate over and over again until it finally reaches a certain critical mass of
bacteria, microbes in the body. Then all of a sudden, we get the signs and symptoms
of an infection. As it begins to multiply in the body, one cell to two, eventually to
millions, it doesnt want to make its presence known. B/c then were going to treat
it, were going to use some antibiotic it, some drug, and were going to quarantine
ourselves. For the bug thats bad. Its either going to be killed or its not going to be
transmitted from one person to another. It waits until critical mass and then the
signs and symptoms of an infection occur. Then its going to take a while for us to
treat it and get better once it gets to a certain concentration in the body. Damage,
the agent and the host response can cause damage. What does that mean? We are
the host. Can we cause damage to ourselves? Yes we can. One of the best examples of
that we found w/ periodontal disease. Well have some information on the microbes
involved in periodontal disease as well, but its our bodys response to the bacteria
that are trying to destroy all the tissue around our teeth. The periodontal tissues
that hold the teeth in place. Our body response is to try to keep the infection in the
area localized. Prevent those bacteria causing that damage from getting to the blood
stream b/c they get into the blood stream and travel through the body, almost any
internal organ could be affected and often poorly so try to maintain that infection
there. Sometimes our body can over respond to keep the infection localized. The
response of the system thats overdone in a way causes as much damage as the
bacterium itself. Our host response/ immune response to localize the infection often
contributes to the damage done to the tissue esp. in periodontal disease. Another
one would be TB as well. Our immune response sometimes contributes to the
severity of the infection by causing more damage than the bug itself would. Outcome
either the agent or the host wins or they learn to coexist.
[11] [Contd]
8

Transcribed by Amit Amin

10/31/2014

[Dr. Boylan] The manner in which each agent combats host defenses distinguishes
one parasite, one infectious agent from another. Its not just one bacterium that
causes staph infections and TB and diarrhea and other disease were going to talk
about. Theyre different bacterium. They have different virulence factors. Its not
just S. aureus causing these things why does it cause the types of infections it does.
B/c of the virulence factors it has that contribute to the pathogenicity of it. They
have certain ones, it has certain ones, and S. aureus does. E. Coli has some others
that cause damage. The TB bacterium has other virulence factors so were going to
have to pay attention to when we talk bout infections, what are the virulence factors
that these bacterium have that cause caries, periodontal disease, buccal mucosal
infections, that causes infections elsewhere in the body. What is that unique
combination of factors that they produce theyre willing to become pathogenic. Each
of these steps requires the breach of host defenses as we pointed out.
[12] [Encounters]
[Dr. Boylan] Endogenous infections caused by known microbiota present in our
own bodies, skin, mucous membranes, weak sites where they are not normally
found. Colonization vs. infection. Theres often some controversy among
microbiologist about the definition of these two terms. I prefer to say we are
colonized by our normal microbiota as they are growing on our skin/ body they are
colonizing. They are forming little micro colonies. They are not causing any damage
for the most part. You saw the benefits that we have as a result of them being there,
being colonized, growing multiplied colonies of 10, 20, 30, maybe larger numbers of
bacteria/ colonies sticking together. Then I say infection is when you are exposed to
some new microbe thats not ordinarily found in our body. Ordinarily found in our
body. Then we become infected w/ this other microbe and if it gains a toehold in our
body and begins to grow, reach a critical mass, it can cause a disease. Some
pathological damage to our tissue (disease). Some people I will mention one
persons name and I can see whose point, said that we are infected from birth. That
our normal microbiota is really an infection that we have and that we get sick from
it when we have an infectious disease. Dr. Caulfield says that. Weve had our
discussions about that. I believe we are colonized from birth, he says we are infected
from birth. So when he taught this lecture in this course or another course dont be
confused. We try to agree on the definition of terms in different faculty in the course
of course but some people say no this is what it is. We are infected, no we are
colonized. So, be aware of that. Colonization, are normal microbiota colonizes us,
infection either vary in the right circumstances may breach our immune/ defense
response. They can cause infection. An infection ordinarily means some microbe
thats not ordinarily in the surrounding, we are exposed to it and it causes disease
and pathological change (he breaks up his sentence often and starts another one).
[13] [Encounters]
[Dr. Boylan] Exogenous infections, contracted form exposure to microbial agents in
the environment. TB, diphtheria, most of the ones we will talk about in the course
will be from an exogenous source. We will be exposed to them by the airborne route,
by the eating/ drinking fluids, fluid contaminants of bacteria, by sexual intercourse,
9

Transcribed by Amit Amin

10/31/2014

by other ways the bacteria will seek to get into the body. Some infection occurs from
somewhere else. Wed rather get contact or exposure to some other source of the
agent, we get sick. Thats exogenous as opposed to indigenous infection. An example
is you catch a cold, eat contaminated food, typhoid fever is one but there are many,
most of these we will discuss in the course are exogenous infections.
[14] [Routes of transmission]
[Dr. Boylan] Routes of transmission. Vertical transmission, parents to offspring.
Generally mother to fetus or infant transplacental, birth canal, breast milk. So, as I
said, ordinarily the fetus is sterile, but sometimes some pregnant come down w/
infections and a microbe can be transmitted from the pregnant woman to the fetus.
One of the best examples of that is syphilis. That can be very hazardous to the fetus
that is exposed to this spirochete, this bacterium that causes syphilis. The fetus is
going through the growth stages trying to develop all of its organs, hearts, lungs,
skin, and brain and all of a sudden if it gets infected at that stage in the womb by a
spirochete or some bacterium the outcome is usually not very good for the fetus if
they become infected. As a competition, the spirochete wants to use a fetus to grow
as a source of nutrients so there is a couple other but we will talk about them later.
The birth canals, so the transmission of the microbes from the mother to the child is
called vertical transmission. Breast milk as well. So ordinarily its mother to child.
There is at least one infection I know of thats transmitted from father as well as
mother to child. That is Hep. B. Hep. B can be transmitted by vertical transmission
from parent to offspring by either mother or father that happens to harbor the virus
that causes Hep. B. Remember that.
[15] [Routes of transmission]
[Dr. Boylan] Horizontal transmission, I thought these were competing pictures.
This one I ordinarily use and I found this one as well. Horizontal, from one person to
another. Not vertical but horizontal. Most of the infections are transmitted in that
way. Direct contact, aerosols, and fomites, arthropod bites. Do you know what
fomites are? Can you transmit an infectious agent by a fomite? Yes you can. A fomite
is really an innominate objet like a doorknob, the seats youre sitting in now. It is
possible that some of you have a cough or the flu you can cough it up in your hand,
touch the doorknob seat, and later on you can touch that same site and pick up the
infection yourself. The infectious agent, touch your eye, touch your nose, touch your
mouth and the microbe gets entry into your body. Its an inanimate object, in
hospital settings, the bedding, the sheets, the pillowcases, the towels, are often
sourced in them. Lab coats are often sources. They have the microbe there and thats
one of the sources I think but well see it can be transmitted from one person to
another. Arthropod bites, fleas, ticks, etc. Respiratory route. Salivary, GI spread,
fecal-oral spread is another common way of transmission of especially food bourn
infections. Fecal oral spread. What does that mean? Typhoid fever is one of the best
examples of fecal-oral spread. Typhoid Mary, well tell this story and take a break,
was a cook who lived in NYC area about a 100 years ago. She came from Ireland and
she harbored the bacterium that causes typhoid fever, which is a very serious
infection. High fever, high mortality rate, very common infection 100 years ago in
10

Transcribed by Amit Amin

10/31/2014

NYC. We didnt have sanitation like we have today. She worked as a cook. There
were a couple outbreaks of typhoid fever among the family she worked with and
finally they traced her down and they found out that her gallbladder was almost
100% colonized by the bacteria that causes typhoid fever and she was shedding
these bacteria in her gut going to the bathroom, not practicing good hygiene and so
go to the bathroom, your fingers, your contaminated w/ a bacteria and then you
start to cook and have handle food for your families and then the transmission
would be fecal-oral way and they would eat the food and the bacteria would be
there. Thats commonly why they say was you hands before you go back into the
kitchen or handle food. Fecal-oral transmission is a very common mode of
transmission of bacteria. They finally tracked her down after some deaths, the
health department, she wasnt the only one who had typhoid fever. She was the only
one who carried the bacteria but the problem was that when they tried to arrest/
quarantine her she put up a terrible fight. She was a tough Irish woman and she
struggled w/ the policeman apparently and some local photographers from the local
papers got a picture of her fighting and thats how she became so notorious for a
carrier of typhoid. She was never sick herself from typhoid fever so she didnt
believe she was a source of the outbreaks of typhoid fever. They arrested her, they
released her if she promised to never cook again. Guess what, she says Ok, I will.
Never cook again, I mean for families. She worked in institutions and hospitals. Ok,
no more. Also come back and report every three months to the health department.
Ok. Well she promptly disappeared. Went back to work. She only knew how to cook.
She had to make a living, and thats all she knew how to do. Her problem was that
she was a very good cook and then there were more outbreaks of typhoid fever.
They tracked her down again and they put her on an island up here in East River on
the tip of Manhattan called Brothers Island and they spent the last 20 some years of
her life on this island so she couldnt be the source of transmission of the typhoid
fever bacteria to other people. They also said, look if we take out your gall bladder
well let you go. No, I want my gall bladder. What are you talking about, Im not sick.
Ya youre not sick but youre a carrier. Fecal-oral spread of the bacteria. Blood-blood
borne. You get infection from the blood-borne route. Genital secretions, of course
sexual contact. Eye-to-eye. And zoonotic infections are those that are derived from
some animal source. Some animal thats sick or carries a microbe that maybe to
itself is not hazardous but we eat the eat or drink the milk of the animal and we will
get sick from that animal. Zoonosis, infection transmitted from animals to humans.
Lets take a 6-minute break and well pick up the next hour.
Ok. Pick it up and progress through these slides rather rapidly I think. Ok now,
overview of the infectious disease process. Properties that microbes have, where
they are found, how theyre transmitted. Were going to get through some of the
properties that they have. The virulence factors they produce and they will cause
infections.
[16] [No title]
[Dr. Boylan] Different ways of transmitting infections shown here. Direct methods
above the blue line here. Hand or object contact, respiratory droplets, contact w/
11

Transcribed by Amit Amin

10/31/2014

animals. Contact. Here for the most part you have other innominate objects, indirect
methods of transmission of an infectious agent for me to you. Some intermediary
was involved. The food was contaminated and then you ate the food, water, cholera
is spread that way. Contact w/ objects. Fomites that might have microbes on them
from some other source or person. Ingestion of contaminated soil. Soil w/ bacteria.
Tetanus is a prime example of that. Arthropod bites. Can anybody tell me some
arthropods that can help transmit infections in humans? Mosquitos. How about
Lymes Disease? Ticks. How about the Black Plague? Fleas. Well talk about some of
those infections later on.
[17] [No title]
[Dr. Boylan] What we went through, you dont have to memorize at this stage the
different infections shown in the bottom boxes here. Weve gotten through these
methods already. Disease transmission. This will be one here on your slide from
different ways of getting it. Directly as shown here.
[18] [No title]
[Dr. Boylan] The next slide, indirect transmission. Fomites, etc. Water, foods,
aerosols sometimes and vectors.
[19] [Entry]
[Dr. Boylan] Entry. Ports of entry. Examples: respiratory route. How do the
microbes get into our body from some exogenous source. Respiratory route, aerosol,
we inhale them. Digestive route, travels diarrhea for example. Usually drinking
contaminated water w/ E. Coli from some other person. Goneherra, from sex, sexual
contact. Genital urinary tract. The skins cut wounds, surgery, and burns, insect bites
into abscesses. These are different portals of entry of microbes into our body.
[20] [Spread]
[Dr. Boylan] Spread. Usually thats the spread from the site of entry to surrounding
tissue. Staph and strep infections are known to do that. Cause abscesses, present on
our skin, cut/ lesion/ injection of something leads to an opening in our skin.
Breaches our skin so that the bacteria can multiply and spread to underlying tissue.
Dissemination via bloodstream to distant sites, infection of vital organs. As I said,
ordinarily the blood stream is sterile. We have our immune system. There are
neutrophils, macrophages, other white blood cells, part of our immune response so
in case our blood stream are infected even after dental treatments, tooth brushing, if
you have any bleeding there at all, some of the bacteria in your oral cavity will go
into your blood stream. We have defenses. We have sentinels, WBCs, neutrophils
especially looking for foreign invaders. Things that shouldnt be there. We want to
do that so that the microbe is not disseminated throughout the body. If the microbe
starts to grow in the blood stream and multiply it can cause a sepsis or even septic
shock or a persons organs will get infected, get shut down, stop working,
hypotension, etc. Well talk about septic shock later and that can be a very serious
type of infection. We try to prevent the bacteria or the virus from spreading in the
blood stream to large numbers by our immune system. If some bacteria grow very
12

Transcribed by Amit Amin

10/31/2014

well and multiply in our blood steam to very large numbers causing sepsis,
contamination of our blood stream and other internal organs could be infected,
colonized and infected, and damaged/ destroyed. Function lost. Microbes have
evolved ways to overcome defenses of the host to spread in the body. Must
overcome the host immune response so our immune system really comes into play
to defend us against infections.
[21] [Other terms]
[Dr. Boylan] Some other terms we use in infectious disease you need to be aware
of. Nosocomial infections. Where are they ordinarily found? Ever heard of the term
nosocomial? Very first two infections acquired in a health care settings. Primarily
hospital settings. People go in for surgery they are healthy otherwise. They come
down w/ infections when they are patients in the hospital. Those are called
nosocomial infections. Most of the time are caused by S. Aureus, E. Coli, and a couple
of others well talk about also. So those infections derived in a hospital environment
or a health care setting as well are called nosocomial give than term. Contrast to
that we have our Community-Acquired (CA) infections. These two terms differ,
nosocomial in the hospital and outside in our environment in the community where
we all live are called community-acquired infections. WE see now that infections
known as MRSA are becoming to be recognized more and more in the community
environment (CA). MRSA. What does the M stand for? Methicillin. R? S? S. Aureus, a
very dangerous modern problem. Community acquired, methicillin resistant S.
Aureus. We use to think these MRSA were only found in a hospital environment.
That was where the danger was. People going for surgery or other conditions to a
hospital. They are already not 100%. Therefore some reason, immunocomprimsied
sometime. They are exposed to the MRSA which is S. Aureus which can cause very
serious infections and are resistant to many antibiotics but now we see more and
more people are being infected w/ MRSA out in the community not in the hospital
setting. Particularly athletes who use locker rooms and theres a lot of bleeding and
injuries. Football, you can imagine, knees and bleeding, and you run into a locker
and youre sharing towels. More and more MRSA in the community (CA). .Risk
factors. This is especially important to try to remember for the infections we
discuss. Immunocompromised are more prone. There immune system is comprised.
They are immunocompetent they are immunocomprimised. T/ B cells and other
parts of their immune system are not functioning robustly. They are being
diminished. HIV, well see that later on people harbor the HIV and their T cells are
destroyed by the replication of this virus in them. The T cells, part of the general
really in our system. Once the T cells are destroyed theres nothing there to help
regulate our immune response to infectious agents so these people are
immunocompromised and are susceptible to many types of infections by bacteria
ordinarily thought to be harmless but when your immune system is not working
everything can be dangerous. Every type of microbe in your body. Body radiation,
these are things that put people in risk for infections. Body radiations,
chemotherapy, and immunosuppression for transplants. Steroids to control
inflammation. Who else is at risk for infection more so than normal population.
Infants who dont have a well developed immune system yet. Elderly, whose
13

Transcribed by Amit Amin

10/31/2014

immune system is diminishing/ waning w/ time as you get older. Our immune
system is not as robust as it was when it was younger. In stage alcoholics, drug
attics, cancer patients, . These are conditions that put patients at risk of other
infections.
[22] [Multiplication]
[Dr. Boylan] Multiplication and incubation period. So rarely is the host infected w/
a sufficient large number of microbes at the initial encounter to cause an infection.
We talked about that before. They must multiply in the host until they reach the
critical match. After we expose to an infectious agent, they are using as an incubator.
They are growing, multiplying in us until they reach a certain number in our body,
the critical mass. Then you being to see the signs and symptoms of the infections.
That period b/w the exposure to the microbe and the time we come down w/ some
symptoms of the infection, its called the incubation period. Incubation period. Time
b/w encounter and the additional symptoms of disease. Were going to talk about
incubations periods of some infections that may be, one day- four days. Or 1 weekweeks. What accounts for the variation of the incubation period in various people.
One of them is the infecting ghost. How man of the microbes are you exposed to. If
you re only exposed to only a 100. Its going to take a while for that critical mass to
be reached. It multiples over and over again via their incubation period. Critical
mass reach, signed and symptoms. If we are exposed to a lot of microbe then the
infecting dose it high. Its going to take a lot less time to reach that critical mass. A
shorter incubation period. Thats one reason the incubation period can very. The
dose youre infected w/. Another is the immune status. How healthy hare you? How
robust is your is your immune response to infectious agents and it varies among
people as well. Of course the virulence of the organism. Some microbes officially
have an incubation period of a day or few days, others might be traditionally be
weeks or months. It depends upon the factors they produce when they produce
when are growing in our body, which enables them to our tissue. How virulent are
they, how dangerous are they. The cold virus for example is one thing. You get a
cold, thats an infection but the flu is another virus thats much more dangerous.
They vary in the virulence factors they produce that also counts for how long or
short the incubation period is. Communicability during. Can infections be
communicable during the incubation period before you start to feel sick? Yes,
definitively. Thats the hazard once again of microbes being sneaky as you can say.
They dont want to make themselves known, present too soon b/c youll treat them
using drugs, youll quarantine yourself, youll stay home. They want to keep on being
transmitted from you when youre getting sick to somebody else. So even before you
know youre coming down w/ an infection your incubation period, you can transmit
these microbes to another person and youre much more likely to do it in many
infections b/c youre not aware that youre harboring these transmissible agents,
you wouldnt learn that staying home, staying home, taking antibiotics for other
therapies. So often, many infections are transmitted during the incubation period
esp. if you have infections that are secreted. Some more than others are transmitted
through the incubation period. Talk about the Ebola virus. Incubation period can be
up to 10-21 days. Or through the 21 days actually they were saying. It depends on
14

Transcribed by Amit Amin

10/31/2014

how robust the immune system was, the infecting dose, etc. They also say its not
until you get the fever that youre contagious. W/ Ebola you do have a symptom of
an infection. You get the fever. During whatever 2-4 days before you get the fever,
apparently Ebola is not transmitted during that incubation period. Only after you get
the fever. When you get the fever, the incubation period is over and then they say
Ebola is transmitted after that. Not before. It can vary. Some infections once again,
transmitted much more routinely during the incubation period that other.
Especially shed in secretion as shown there.
[23] [No title]
[Dr. Boylan] Lower course of infection. Here we have someone coming down w/
the flu. Some other terminology used. Day 0 for this child here is exposed to the flu
virus from the mother or someone else in the community through the aerosol route
(coughing, talking, sneezing). It is spread, the person inhales the flu virus and the
incubation period exposed to day 0. Entrance of the virus in the susceptible person.
Prodromal phase, thats when you kind of feel youre coming down w/ something.
Youre getting a fever, something is coming on. Minor aches or pains. Runny nose.
Nothing really thats significant yet in the prodromal period. But then the incubation
period is over. When you did come down in the case of flu w/ high fever, chills, joint
aches, tiredness. Thats the acute phase/ period of the infection. Once again, all
infections follow this. Especially exogenous infections follow this pattern. The length
of these days or intervals of these different periods may vary quite a bit but its
more or less the same progression after exposure to the signs and symptoms till you
start to get better. So then the flu will have these signs and symptoms. Fever, chills,
joint aches, and really body aches, another symptom of the flu that you find around
is a dry cough. You dont cough up spulent from your lungs but your coughing a lot.
A dry cough, fever, and body aches are typical of the flu. Much more severe
symptoms than you find w/ the common cold caused by a different virus. Then you
start to get better as your immune system kicks into play. The period of decline.
Then after day 11 or so, in the case of flu youre pretty much back to normal. Period
of convalescence, youre getting rid of last particles of flu virus in the body. Your
immune system is still acting to neutralize and keep the virus under control so it
doesnt cause another infection. Recurrence of the infection and then you get better
hopefully. So those are the stages of an infectious disease . Any, not just flu. Almost
any infection we talk about has to go through these stages.
[24] [Damage to host]
[Dr. Boylan] Damage to host. Now well talk about the pathogen. The microbe that
causes the infectious disease. Produces the virulence factors by toxins, enzymes,
invasive factors that destroy the tissues of the host. Some pathological change of the
tissues is caused by the result of these microbes growing in us. The tissues infect
depend upon a particular microbe. Additional damage to the tissues done by the
host vehement response. TB, we mentioned that before and periodontal disease.
[25] [Progression and outcome]
[Dr. Boylan] Lets take a look at your ledger of the progression and outcome of a
disease. Potential pathogen. Different things could happen. It approaches us. It gets
15

Transcribed by Amit Amin

10/31/2014

on us. We can say OK, we like you. Youre not going to cause damage to us. In this
case here, going to the left, transient microbiota, sometimes they just pass right on
through us. They are transient. They travel through us. They dont do damage at all.
Others can bind to us somewhere. Become part of us, commensals and later on
cause an infection. Others, no loss, no entry. Leads to loss of pathogen and no
damage. Here you have an infection when the crossing of surface and entry into
sterile tissue results. Some breaching of our immune system. Damaging our tissue.
Getting into our body. Infection occurs. Disease, death and other outcomes of
infection are shown here. A chronic disease is one that lasts for a long time like TB.
In contrast to that we have acute infections, which come on very rapidly like the
common cold. They come on for a very short incubation period, last for just a few
days or even a flu. An acute infection and then it goes away after a couple weeks. TB
and those caused by fungi can be chronic. Long incubation period, long time youre
sick and if you start to get better, it takes a long time to get better. Chronic diseases.
[26] [Virulence Factors]
[Dr. Boylan] Virulence Factors. Heres an overview of what were going to talk
about next. These are the main weapons that microbes have that cause the different
types of infections that they do in us. Capsule, adhesions- essential that they stick to
some site/ cells/ tissue in our body in order for them to grown, multiply , and
eventually cause an infection. Invasins are the enzymes that many produce. Toxins,
two types, exo-/ endotoxins. Antigen switching, well talk about that later as well.
They can change their antigen on their surface so our immune system dont
recognize them and many bacteria have some novel virulence factors as well such as
the TB bacterium which has a waxy cell wall which is its main virulence factor
allowing it to cause infections in humans. This is just a partial list but its pretty
much an overview of what well be talking about and the various infections well
discuss. What are the virulence factors that particular bacterium/ virus has that
enables it to cause that particular infection. A skin infection not a lung infection. A
liver infection not a GI infection. What are the virulence factors they have.
[27] [Capsule]
[Dr. Boylan] First of all theres the capsule. Capsule also known as glycocalyx for
sweet coat. Also known as the slime layer since its slimy, its viscous,
polysaccharide layer that surrounds the bacteria as you may recall from the course
of microbiology. Viscous consistency. Its shown here. Here we have bacteria, its a
chain here. The dark cells are streptococci. Streptococcus. Probably a pneumococcus
b/c you see a lot of pairs but the white area around it is the capsule, polysaccharide,
glycocalyx, a tremendous virulence factor for bacteria that have it. It helps the
bacteria that are found here that are enshrouded w/ it from being phagocytized.
They get into the body, they have a capsule. Here we have our neutrophils looking
for these bacteria that shouldnt be there in our tissues, lung, and blood. But they
cant grab a hold of bacteria that are here. They cant grab a hold of them b/c they
approach the bacteria and the capsule protect these bacteria from taken up and
ingest by our neutrophils and macrophages and they slip away. Avoiding
phagocytosis is a very important virulence factor a bacteria can have and the
16

Transcribed by Amit Amin

10/31/2014

capsule is the primary one that enables bacteria to avoid it. Its not found in all
bacteria. Its not found in all pathogens but bacteria that do produce infections like
pneumonia and meningitis, most of them do have capsules. They have luxurious
capsules as shown here. Thats a large capsule. What kind of stain is this anyone
remember? We use often to show if bacteria have capsules. Not a gram stain, simple
stain, but a, youre thinking about it, its almost there and? A negative stain.
Remember a negative stain. It doesnt stain the bacteria or capsule but it stains the
background more or less. Also, capsule prevents against desiccation. Think of it.
These bacteria, they have a mucoid polysaccharide component. If somebody has
pneumonia coughs up the sputum and it lands here on this top bench here/
doorknob and if the bacteria dries up right away, they wouldnt be pathogenic, they
would die w/in a short period of time. Desiccation is bad for bacteria that are out
there in the environment. If they have a capsule surrounding them, this capsule
kinds of protects the bacteria inside the capsule material from drying up. Being
desiccated. They can survive longer. If they can survive longer outside the body,
theres a better chance someone will come around eventually and contact them on
some fomite and then come down with the infection. Also, its not the main
component for adherence, capsules and some bacteria especially this one since its
on the surface and help them adhere, stick to our tissue. Well see that Pilli are the
main sources of adhesion of bacteria. Capsule can also play that role. Help the
bacteria, the capsule part of the bacteria can help them often stick to ourselves as
well. Its a very important virulence factor. Never underestimate the power of the
capsule to be a virulence factor for bacteria. Especially those that cause pneumonia
and then meningitis. Theyre tough to get rid of b/c this capsule protects them from
our innate immunity and it helps them slip away and not be digested, survive and do
more damage in our body.
[28] [Adherence]
[Dr. Boylan] Adherence in this new capsule. Components of bacteria that help them
adhere are called adhesions (notice the O) they help in the adhesion (the process)
but the adhesions are the Pilli that bind to receptors on our cells. Pilli are also
known as fimbriae. Lipoteichoic acid if you go back to look at your notes from
microbio youll see LTA its only found in gram-positive bacteria. It sicks up through
the peptidoglycan. It can also help the bacteria adhere to the tissue. The M protein of
streptococci, well talk about that when we talk about streptococci next week.
Especially the bacteria that cause strep throat. Streptococci that cause strep throat
have this factor called M protein on their surface that helps them adhere to our
tissue that then eventually causes strep throat. Capsules we talk about before. They
enhance virulence by, once they adhere/ stick theyre not carried/ swept away from
mucus or the flow fluids for example the urinary/ GI tract. They are able to hang on
to our tissues. These (some word) adhesions here, they can adhere to every tissue in
the body thats adhesions thats on the surface of the body. They look for receptors
on our cells. E. Coli in our gut. They find receptors in our gut. Diphtheria bacteria
find receptors on our cells in our throat, etc. These adhesion bacteria look for
receptors. They have to bind to something. Adhesion and receptor. Adhesion on
bacteria and receptor on our cells and then they begin to grow and colonize and
17

Transcribed by Amit Amin

10/31/2014

eventually cause infection. They will hang on even when urine is flowing, even when
the peristalsis of the gut, fluid passing through the gut. They can adhere and stick,
not get washed away.
[29] [Invasiveness]
[Dr. Boylan] Invasiveness. Some bacteria, well say a lot of bacteria actually
produce enzymes that enable them to invade through our tissue. Invasions, they are
enzymes spread through the tissue. One of the ones is coagulase. These are just a
few. Coagulase, well talk about 3 different invasins. Coagulase, streptokinase, and
hyaluronidase. Three examples of invasins. These are each enzymes that help
certain bacteria cause infections. Coagulase, here on the upper left here we have the
pathogen that is green. Here is the bacterium in green. It is producing this enzyme
invasin called coagulase. We have blood here. As the result of this activity of the
bacteria producing coagulase, you notice that the bacteria that are shown here in
the middle slide that are still in green are surrounded by something. A fence has
been built around them. Its a fibrin fence. They form a clot. Coagulase, meaning
coagulation, coagulase, thats an enzyme and its produced by our friend S. Aureus
only. S. Aureus. Just this enzyme that causes fibrin clots. Bacteria are fenced in by
this clot. What good is that to the bacteria, this fence around it? Bacteria are hidden
inside this fibrin clot from our immune system. Heres our immune system. Were
looking for this bacteria trying to destroy them. Here the bacteria are winning on
the system by shielding themselves in this fibrin clot so our immune system wont
recognize and destroy them. In the middle of this clot, blood clot around the
pathogens, so coagulase does that. Protects the bacteria, therefore they survive, they
continue to grow, divide and cause more damage as times goes by. In contrast to
that, this is the interesting thing about these two enzymes, coagulase vs.
streptokinase. This is another enzyme but it helps the bacteria cause an infection by
just the opposite than coagulase. Streptokinase is an enzyme produced by the
streptococci. Coagulase by staphylococci only staphylococcus origins (only S. Aureus
produces that invasin). The streptococci that genus, produces an enzyme called
streptokinase. This enzyme, here we have the other bacteria, now streptococci in
green on the right here, they produce streptokinase and this enzyme dissolves the
clot. Streptococci dont want to be meshed in a fibrin clot. They want the clot to be
dissolved and they produce and enzyme that dissolves the clot. What good does that
do to the bacteria? Now these bacteria can spread. They are not fenced in as they
are w/ staph in a fibrin clot. They want to break out of the fibrin clot so they can
spread in the blood stream. Spread to underlying tissue, dermis, epidermis, and
subcutaneous tissue. These two enzymes are both virulence factors of these bacteria
but they act in different ways. Heres another one. Hyaluronidase. We know that
there here are epithelial cells here, here, and here bound to each other very tightly.
Heres a pathogen that wants to get through the epithelial layers of our gut, our
mucus membranes and get inside the body. But the cells are so tightly bound
together they cant do it. What do these bacteria do? They produce and enzyme
called hyaluronidase. The kind of cement/ glue b/w our cells and our epithelial
layers is hyaluronic acid. These bacteria want to penetrate these tissues, destroy the
bonds that are linked b/w the adjacent cells. I want to destroy the cement/ glue, the
18

Transcribed by Amit Amin

10/31/2014

hyaluronic acid. The enzyme that destroys the hyaluronic acid is hyluronidase. They
can spread now in the body. This particular enzyme is called the spreading factor
but it helps the bacteria destroy our tissue, the bond b/w our cells, get into the body
and spread to the underlying tissues, the subcutaneous tissue and you eventually
get into o the bloodstream. Heres pathogen, heres the enzyme resolving the
hyaluronic acid, easy access to the underlying tissue. These are examples of
invasins. Invasins are generally enzymes, that microbes produces that help cause
and infection.
[30] [Exotoxins]
[Dr. Boylan] Exotoxins. Exotoxins as the name indicates are toxic, are proteins
produced by the bacteria that are toxic to ourselves. That damage ourselves.
Pathological damage to our cells by toxins. They are called exotoxins b/c the
bacteria released them. They form them inside, in their cytoplasm but then they
kind of push them out through the membrane/ wall. They release them so they can
act at other sites than where the bacteria are found. Both gram positive and gramnegative bacteria can produce exotoxins. The other type of toxin the bacteria have
and can mention later is not the exotoxin but the endotoxin. Its only found in gramnegative bacteria, the OPS remember. But exotoxins can be produced by both gram
positive and gram-negative bacteria and they are released. They are not a part of the
bacteria. They are released by them. The endotoxin is a part of the bacterial cell.
Proteins secreted by bacteria. Couple examples here. Toxic to our cells. Exotoxins
are generally heat sensitivity. Heat labile. In other words, they are inactivated. Head
labile. Sensitive to heat. These are no longer toxic when they are heated 60 degrees
C for just 10 minutes or so. Thats all. Maybe even shorter periods than that. These
exotoxins are heat labile. Brief heating, not too high of a temperature, not too hot,
for just 10 minutes. Toxoids, exotoxins can be converted to toxoids. When you do
indeed heat exotoxins at 60 degrees C for 10 minutes or so they become nontoxic.
They lose their toxicity. However, they are then converted to toxoids, which were
the toxins w/o any ability to cause any problems in us w/o being toxic and they can
still be used as antigens in vaccines. Well talk about that later as well. So exotoxins
can be treated to remove their toxicity but certainly can be used antigens especially
in vaccines like diphtheria and tetanus you use the toxoids. You get the toxin in
these bacteria and destroy the toxicity but keep it around so that the part thats
antigenic is still there and can use in vaccines. Many exotoxins are referred to A/B
exotoxins and I want you to remember what these are. A/B exotoxins. We should
have it on the right here how these things work. A/B exotoxins are exotoxins that
have two parts an A part and a B part. The A part is a toxic part. The A stands for
active, its the active part, the toxic part. I think Im coming down w/ the flu. I had
my flu shot. Get your flu shot. Everybody get your flu shot? Its a little complicated
but I really like the way these exotoxins work. A/B. A part and a B part. Toxic part. A
for active. Its an enzyme. The B part of them is the part that binds. It adheres. The
adhesion of these exotoxin is the B part. What does the A toxin do? Lets shift now
the right and look at the series of drawings here. Here we have a bacterial cell. A
bacterium and the DNA. Its chromosome and this bacterial cell has a gene, this
chromosome for exotoxin production. Exotoxin is a protein so this is gene, bacterial
19

Transcribed by Amit Amin

10/31/2014

chromosome for its production. So the gene is activated and it produces the A/B
exotoxin polypeptide protein. This then leaves the bacterial cell, its excreted as an
exotoxin and now its searches out one of our cells. Heres one of our eukaryotic
cells. Its one of the cells thats going to be effected by this exotoxin. Here we have
the two part. A/B. You see the B part finds a receptor. The A is the adhesion and the
B is a receptor on one of our cells. Attaches to the receptor. It enters the cell. So you
see here, the whole exotoxin enters our cell by endocytosis and it pushes us through
the membrane and takes some of the cell membrane w/ it. Now its in the cell. What
happens next? Toxin enclosed in the inside portion of the membrane. Pinocytosis,
endocytosis. Doesnt matter. Same thing different site. Then what happens next is
the interesting part. The A/B components of endotoxins separate. The A component
is the active component and the B component is then released from the cell. Once its
in the cytosol we dont need the B anymore. We dont need the binding part. We
want just the A. The A part, the active part, the toxin, the enzyme. What is it going to
do? In every case w/ these A/B exotoxins it does the following. Ok. Lets just see
here. Lets shift back over to the left now. This A toxin in here, what it does, is it
attacks. It attacks the compound in the cell called NAD. Remember NAD? Ill take a
break after this slide. So heres (some word). Its in the cell, its freed up. How is it
going to damage this cell? What is it going to do? It attacks the component known as
NAD. Nicotinamide adenine dinucleotide. You remember that from biochem/
glycolysis. NAD. What it does, is it breaks NAD down to into a two component parts.
Nicotinamide and ADP-ribose so these two are the components of NAD. Then the
enzyme A, takes the ADP-ribose and it throws it on to or attaches it to an essential
protein in the cell. Something this cell needs to survive. Some protein, some enzyme,
something essential to the cell. Something this cell needs to survive. This component
of the cell needs to survive, this protein/ enzyme all of a sudden is ADP-ribose and
attaches to it. Its called ADP-ribosylation of a target. So the target is the protein/
enzyme thats essential for the maintenance/ growth of the eukaryotic cell. When
that happens, the cell stops proteins synthesis and the cell dies. So lets go through
this once again. A/B exotoxins. Two parts. The binding part is B, the A part is active
enzyme toxin part. The toxin part is going to kill the cell. The A/B exotoxin injures
the cell and the two parts separate. The B is lost and goes out of the cell. A part what
does it do? It looks for NAD. It cleaves NAD into two parts. The ADP-ribose is one of
the two parts. The ADP-ribose now seeks out a particular protein or enzyme, binds
to it, inactivates it. Stopping protein synthesis and the death of the cell. Ok. This
happens w/ both diphtheria, anthrax, and( one I couldnt understand) some other
exotoxins act that way but the ADP-ribose thats formed in these different infections
picks up different proteins in our cells. For example the ADP-ribosylation would
attach to one protein in our cell. The ADP-ribose causes anthrax infections and some
other protein. Botulism as well. But thats a common feature they all have. The same
thing. A/B. A toxic part. ADP-ribosylation of some essential protein. The essential
protein may vary depending upon the infection whether its diphtheria, anthrax, or
botulism but it shut down protein synthesis. This cell dies. Thats what causes the
damage during the infection by these organisms. Its kind of neat. I dont know how
the ADP-ribose selects for the particular protein it does in an infection. I guess the A

20

Transcribed by Amit Amin

10/31/2014

component must be around guiding the ADP-ribose to a particular protein whether

the diphtheria exotoxin, anthrax exotoxin, etc.
[31] [Endotoxin]
[Dr. Boylan] Thats exotoxin. Gram positive and gram negative cells can produce
them. Endotoxins are only found in gram-negative bacteria. Also known as a
lipopolysaccharide or LPS as shown here. Review of the LPS, you see 3 parts. The Oantigen, core polysaccharide, and the Lipid A. Lipid A. One lipid and two different
polysaccharide parts. The core polysaccharides and the O-antigen, repeating units of
O-antigen. The lipid A is the toxic part of LPS. Lipid A moiety as shown here.. Thats
responsible for the toxicity and the fever during the course of gram negative
infections. The lipid part of LPS. Gram-negative only. Applied to the cell wall,
therefore endotoxin. Heat stable, not destroyed even by boiling. Not converted to
toxoids. So I cannot use endotoxins to form vaccines that we give people b/c not
converted to toxoids as exotoxins are. Heat stable. Not destroyed by boiling.
Exotoxins even by temperatures of 60 degrees C for a few minutes will destroy their
toxicity but not w/ endotoxins. One of the reasons we could be certain about that
was that they use to make up vaccines for people or different types of medications
that had vials, where you picked it up w/ and inject somebody w/ it. This serum, this
pharmaceutical liquid solution we use to administer a drug/ vaccine its sterile so
its safe to give to people. Not true. It can be sterile but if it still has some LPS in it,
not bacteria but certainly LPS from the gram negative bacteria and its contaminated
somewhere on the line, it can cause a lot of serious consequences to the patient who
gets injected w/ the sterile solution that still has LPS endotoxin so now we have to
make sure even anything given in humans has to be LPS free as well. By itself is very
toxic.
[32] [Characteristics]
[Dr. Boylan] Some differences b/w the two. Heres the characteristics of either of
these two. Here are the endotoxin and exotoxin characteristics. Weve already gone
through a lot of these. Endotoxins are LPS. Protein, part of the cell wall, yes. No they
are secreted. Gram positive, no. Only found in gram negative. Extracellular. No.
Endotoxin is part of the cell wall once again. (Mumbles). No, Yes. Many of the genes
for exotoxins are found in different sites in the bacterial cell. Some of the genes for
exotoxins are found on the chromosome. Other genes for other exotoxins are found
n plasmids. And other genes for exotoxins maybe found on bacteriophages.
Bacteriophages infect bacterial cells, those viruses that infect bacteria and the
prophage is part of the chromosome of the bacterial cell. The point being here is that
here is 3 potential sites of genes for exotoxin production in bacteria. Chromosomal,
phage, and plasmid and different exotoxins in different bacteria are found on these
different sites. Antigenic. Even though its called the O-antigen, its a very week
antigen. Where as exotoxins and the toxoids from (some word) are very
immunogenic. No, Yes. Stable to boiling. Yes and No. Know these differences b/c the
two types of toxins bacteria can produce that enable them to cause infections.
[33] [Some properties.. ]
21

Transcribed by Amit Amin

10/31/2014

[Dr. Boylan] Here are just some of the terrible sequence of infection from
endotoxins. I think, wait a minute, our body is coated w/ gram negative bacteria.
They all have LPS. Why arent we being damaged by them? Why arent we coming
down w/ these characteristics shown here. Our immune system recognizes. Our
immune system knows our normal microbiota and keeps them under control.
Doesnt let them grown too extensively causing infection or disease but when some
ne gram negative bacterium comes in where this new different exotoxin comes into
play. Our immune system can differentiate from our normal microbiota and a
foreign invader. These are some of the effects of the endotoxin. Pyrogenicity. What
does that mean? Pyro-fever. The lipid part. DIC- disseminated intravascular
coagulation. Clotting. LPS can cause clots to form in our blood. Especially in our
peripheral organs, fingers and toes. Clotting that means the circulation of the blood
is slowed down. Our blood stream is not functioning/ flowing as well as it could.
Prone therefore to an infection at those distant site where those clots occur. Shock.
It can inactivate our normal organ function. Hypotension. Low blood pressure.
Hemorrhaging. Lower plate/ neutrophil counts. You can see in any gram-negative
infection we discuss, the LPS plays a very important role in the establishment of the
infection by gram-negative bacteria.
[34] [No title]
[Dr. Boylan] This is more or less showing the same thing. Dont worry about this
slide. You can kind of make sense of what weve mentioned. Its different effects of
the LPS. Whats on the previous slide is enough for you to know. Very interesting,
very potent biomolecule, the LPS, endotoxin of gram negative bacteria.
[35] [Virulence]
[Dr. Boylan] How to measure virulence of bacteria? Pathogen. Well go to the next
slide and stop there. The degree of pathogenicity. How you measure whether youre
going to get the common cold or the flu or hepatitis. How pathogenic are these
microbes. What kind of terminology do we use. This is a very serious pathogen. This
is a pathogen but its not as deadly. Not as much more mortality/ morbidity.
[36] [Virulence- LD50]
[Dr. Boylan] Well stop at this one. There are tests that are done in the lab to
determine the degree of virulence of a microbe. How dangerous is it and this
example here is done quite a bit or I should say were trying to use fewer and fewer
animals in these studies. Here were going to test the virulence of a microbe called
the LD50 experiment. LD stands for lethal dose. Lethal dose, what dose of the
bacteria kills the animal. Not just kills the animal. What lethal dose, what dose of the
bacteria causes death of 50% of the animals. LD50, what is the dose and how many
bacteria does it take to kill half the animals in the study? Why dont we say 100%,
well we do this study and we can find some animals not to be killed by even the
most virulent pathogen so its good enough to determine what amount of bacteria
kills half of the animals to determine its virulence. Here we have 5 groups of mice
and we have 10 mice in each group. They are all going to be injected w/ the same
bacterium but at different dosage of the bacterium. Mice in group A, only 100
22

Transcribed by Amit Amin

10/31/2014

bacteria are injected into them. In group B, 1000, 10,000, 100,000, and 1,000,000
bacteria. Each of the 10 mice are injected w/ a different dose of bacteria. You come
back the next day, look at the killed mice of the 10 and how many have died in each
cage. What you see here in group A is that none of the mice died. Its totally not a
lethal dose. 100 is not enough to kill any of the mice. How about in group B? 2 of the
10 mice died. Group C, 5 of the mouse die. In D, 8 and group E, all of the mice were
killed by the dose that was given. So what is the LD50 in this experiment? Right. 10
to the fourth . It killed half of the mice. To do this, you need to you say OK when we
do it we find the same results but lets see what happens if we give these mice an
antibiotic before you inject the bacteria? Lets see how many, if it takes a larger dose
now to kill them so we see there a little usefulness to these studies to determine
how effect a new drug may be or a new treatment to protect these mice from being
killed by a dose of bacteria they were killed by before. Ok, well stop there and well
see you next week.

23