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EBOOKS Effects of Genetic and Pathogenic
FOR THE Diseases onCells

APPLIED BIOLOGY COLLECTION
Christopher J. Paradise A. Malcolm Campbell
SCIENCES
LIBRARY Several genetic and pathogenic diseases are described to
illustrate how diseases can and do disrupt normal molecular
Create your own and cellular functions, and how those disruptions affect entire
Customized Content organisms. In the case of genetic diseases, how they arise and
Effects of
Bundlethe more are maintained in populations is discussed. In the case of patho-
books you buy, genic and parasitic organisms, understanding their complex life
cycles and their modes of transmission is critical to understand-
the greater your
discount!
ing their effects on individuals and how disease outbreaks occur
in ecological systems. Communication between the pathogen
and the host organism occurs in the course of infection and
Genetic and
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Energy Physics
involves the disruption of normal cell function. Finally, epide-
miology is briefly discussed, using the case of severe acute Pathogenic
Diseases
respiratory syndrome (SARS). Data are used to describe how
Engineering
the disease may have originated and evolved to infect humans,
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and how it spread relatively quickly and almost caused a global
onCells
Biology pandemic. Understanding how disease outbreaks occur in eco-
Mathematics logical systems is critical to controlling the spread of disease.
Chemistry
Christopher J. Paradiseis professor of biology and environ-
mental studies at Davidson College. He teaches introductory
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A. Malcolm Campbell
Effects of Genetic and
Pathogenic Diseases
onCells
Effects of Genetic and
Pathogenic Diseases
onCells
Christopher J. Paradise, PhD

A. Malcolm Campbell, PhD
Effects of Genetic and Pathogenic Diseases on Cells
Copyright Christopher J. Paradise and A. Malcolm Campbell. 2016.
All rights reserved. No part of this publication may be reproduced,

storedin a retrieval system, or transmitted in any form or by any means
electronic, mechanical, photocopy, recording, or any other except for
brief quotations, not to exceed 250 words, without the prior permission
of the publisher.
First published in 2016 by

Momentum Press, LLC
222 East 46th Street, New York, NY 10017
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ISBN-13: 978-1-60650-961-6 (print)

ISBN-13: 978-1-60650-962-3 (e-book)
Momentum Press Biology Collection
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Printed in the United States of America

Abstract
Several genetic and pathogenic diseases are described to illustrate how
diseases can and do disrupt normal molecular and cellular functions, and
how those disruptions affect entire organisms. In the case of genetic dis
eases, how they arise and are maintained in populations is discussed. In
the case of pathogenic and parasitic organisms, understanding their com
plex life cycles and their modes of transmission is critical to understanding
their effects on individuals and how disease outbreaks occur in ecological
systems. Communication between the pathogen and the host organism
occurs in the course of infection and involves the disruption of normal
cell function. Finally, epidemiology is briefly discussed, using the case of
severe acute respiratory syndrome (SARS). Data are used to describe how
the disease may have originated and evolved to infect humans, and how
it spread relatively quickly and almost caused a global pandemic. Under
standing how disease outbreaks occur in ecological systems is critical to
controlling the spread of disease.
Keywords
mutations, disease, point mutation, motor neuron disease, familial disease,
normality, emerging disease, cytokines, rice blast fungus, pandemics, sickle-
cell disease, epidemiology, perfection, pathogen, vector-borne disease, Lyme
disease, macrophage, neutrophil, parasite, Giardia, outbreak, SARS, disease
reservoir
Contents
Preface...................................................................................................ix
Acknowledgments....................................................................................xi
Introduction.........................................................................................xiii
Chapter 1 Genetic Diseases Affect Cells and Organisms.....................1
Sickle-Cell Disease is Caused by a Single Point
Mutation........................................................................1
Multiple Mutations may Cause a Single Disease.................9
Ethical, Legal, Social Implications: Normality
andPerfection...............................................................12
Chapter 2 Pathogens Affect Cells and Organisms.............................15
Lyme Disease Is Caused by a Vector-Borne Bacterium......15
Ethical, Legal, Social Implications: There are Several
Issues with Using Animals in Research..........................22
Fungi can be Pathogens of Plants.....................................24
Chapter 3 Parasites can Survive in More than One Host Species.......29
Chapter 4 Diseases Spread Through Populations of Susceptible
Hosts...............................................................................41
Conclusion............................................................................................51
Glossary................................................................................................53
Index....................................................................................................55
Preface
This book about the effects of genetic and pathogenic diseases on cells is
part of a thirty book series that collectively surveys all of the major themes
in biology. Rather than just present information as a collection of facts,
the reader is treated more like a scientist, which means the data behind the
major themes are presented. Reading any of the thirty books by Paradise
and Campbell provides readers with biological context and comprehen
sive perspective so that readers can learn important information from a
single book with the potential to see how the major themes span all size
scales: molecular, cellular, organismal, population and ecologic systems.
The major themes of biology encapsulate the entire discipline: informa
tion, evolution, cells, homeostasis and emergent properties.
In the twentieth century, biology was taught with a heavy emphasis
on long lists of terms and many specific details. All of these details were
presented in a way that obscured a more comprehensive understanding.
In this book, readers will learn about the effects of genetic and patho
genic diseases on cells and some of the supporting evidence behind our
understanding. The historic and more recent experiments and data will
be explored. Instead of believing or simply accepting information, read
ers of this book will learn about the science behind the effects of genetic
and pathogenic diseases on cells the way professional scientists dowith
experimentation and data analysis. In short, data are put back into the
teaching of biological sciences.
Readers of this book who wish to see the textbook version of this
content can go to www.bio.davidson.edu/icb where they will find
pedagogically-designed and interactive Integrating Concepts in Biology
for introductory biology college courses or a high school AP Biology
course.
Acknowledgments
Publishing this book would not have been possible without the generous
gift of Dr. David Botstein who shared some of his Breakthrough Prize
with co-author AMC. Davids gift allowed us to hire talented artists (Tom
Webster and his staff at Lineworks, Inc.) and copyeditor Laura Loveall.
Thanks go to Kristen Mandava of Mandava Editorial Services for project
management and guidance. In particular, we are indebted to Katie Noble
and Melissa Hayban for their many hours and attention to detail.
Kristen Eshleman, Paul Brantley, Bill Hatfield and Olivia Booker
helped us with technology at Davidson College. We are grateful to ad
ministrators Tom Ross, Clark Ross, Carol Quillen, Wendy Raymond,
Verna Case, and Barbara Lom who had confidence in us and encouraged
us to persist despite setbacks along the way.
Thanks to my wife Amy Brooks for her constant support during the
development of this textbook, and my daughter Evelyn for her endless
energy. Thanks to Malcolm Campbell for his steadfast resolve and opti
mism. Without him, this book would not exist. Thanks to collaborator
Laurie Heyer for taking my sometimes half-baked math ideas and turn
ing them into powerful and elegant Bio-Math Explorations. I learned
a lot from both of them. While the math is largely absent from this
book, our collaboration with her made this a better book. Nancy Stamp
at Binghamton University, and Bill Dunson and Richard Cyr at The
Pennsylvania State University influenced me greatly in how I think as
a scientist and approach my teaching. Finally, I thank my students in
Integrated Concepts in Biology II, who enthusiastically participated
in our experiment to redesign introductory biology, starting with the
text and ending with a new approach to teaching biology.
Introduction
In the latter half of the seventeenth century Anton van Leeuwenhoek
used a simple microscope to become one of the first people to view liv
ing cells in the form of blood, rainwater, and algae. He viewed microbes
and cells of multicellular creatures. Much later, in the mid-nineteenth
century, other scientists observed plant and animal tissues under micro
scopes. These scientists discovered that the parts are made of cells and
that cells could be considered individual units within multicellular organ
isms. This led Henri Dutrochet to state that the cell is the fundamental
element of organization and Rudolph Virchow, a German physician, in
1855 to state that all living cells come only from other living cells. The
observations of these scientists led to the development of Cell Theory,
which states that all organisms are made from one or more cells, cells
only arise from preexisting cells, and the cell is the smallest form of life.
Themes of the cell concept also include: maintenance of an internal en
vironment, structure defines function, and communication. Here how
diseases can have a variety of effects in organisms will be examined. A
single population of a unicellular pathogenic organism can interact with
cells or tissues in one or more host species, or genetic diseases can affect
cells and tissues in multicellular organisms.
CHAPTER 1
Genetic Diseases Affect

Cells and Organisms
Genetic diseases are caused by mutations. Mutations are changes in the

sequence of DNA, so they produce altered RNA and proteins. Altered
proteins may not function properly, causing the cells in which the pro
tein is expressed to not function properly. The functions of a cell are an
emergent property of all the molecules in the cell. Very small changes are
all it may take for a radical change in function to occur. This can lead to
problems for the entire organism.
Sickle-Cell Disease is Caused by a Single Point

Mutation
Many human diseases are caused by genetic mutations. A disease is a
condition of an organism that impairs normal functioning, manifested
by distinguishing signs and symptoms. Most are relatively uncommon
in human populations. The prevalence of a disease is dependent upon
characteristics of the population, the environment, and whether or not a
mutant allele that causes disease is dominant or recessive. An example of
a genetic disease about which much is known is sickle-cell anemia. Sickle-
cell anemia is a blood disorder in which red blood cells assume a sickle
shape, almost like a quarter moon, which is different from the normal
donut shape of these cells.
Sickle-cell anemia affects millions of humans throughout the world,
particularly those whose ancestors come from sub-Saharan Africa, South
and Central America, India, and Mediterranean countries. In the late
1940s, scientists observed that about 8% of the population of African
Americans had red blood cells that sickled when exposed to low oxygen
2 EFFECTS OF GENETIC AND PATHOGENIC DISEASES ON CELLS
pressures. They typically suffered no long-lasting effects from this phe

nomenon. Amuch smaller percentage, about 2.5%, suffered from a severe
form of anemia with many red blood cells sickling and being destroyed.
Those suffering the mild form had about 1% of their red blood cells sickled
in the blood returning to the heart, whereas those with anemia had 30% to
60% sickled. Observations revealed that the lower the oxygen, the greater
the percentage of sickled cells in both of these types of individuals. The ad
dition of oxygen reversed the sickling effect.
The famous scientist Linus Pauling and his colleagues hypothesized
that, because of the connection with oxygen, individuals suffering from
sickling possessed a different form of hemoglobin (a protein that binds
and transports molecular O2) than unaffected individuals. Pauling and
his colleagues performed experiments to compare normal and sickle-cell
anemia hemoglobins using electrophoresis, a technique used to separate
ionic molecules according to their charge.
The researchers placed hemoglobin in a carbon monoxide (CO) solu
tion, leading to hemoglobin molecules fully bound to CO. Bound he
moglobin should exhibit similar properties, whether bound to CO or
oxygen. The scientists prepared solutions of normal and sickle-cell he
moglobin in buffers of different pH. They used a U-shaped tube with an
electric field running through it. The hemoglobin suspension was placed
in the tube and then buffer was added to the top of both ends of the tube
so that it lay on top of the suspension. The boundary between the buffer
and the solution could be observed to determine if it moved relative to
where it started.
Most proteins are negatively charged in solutions of pH that are near
neutral (pH 7). Proteins have different amino acid sequences and have
charges that differ, and this causes proteins to move differently in an elec
tric current. Differential movement of the boundary line at a variety of
pHs would support the hypothesis that normal and sickle-cell hemoglo
bins are different. Finding the pH at which a protein carries no net charge
helps describe the protein (Table 1). Pauling and his colleagues found that
the pH at which there was no movement was 6.87 for normal hemoglo
bin and 7.09 for sickle-cell hemoglobin. Below this no net movement
pH the protein carries a net positive charge, and above this pH it carries
a net negative charge.
Genetic Diseases Affect Cells and Organisms 3
Table 1 Movement of normal and sickle-cell hemoglobin at different pHs.

pH normal hemoglobin sickle-cell hemoglobin
6 positive movement positive movement
6.87 no movement positive movement
7 negative movement positive movement
7.09 negative movement no movement
8 negative movement negative movement
Source: Data from Pauling et al., 1949, Figures 1 and 2.
A way of scanning the protein concentration in an electrophoresis

tube was applied to hemoglobin from normal, sickle-cell anemia, and
mild sickling individuals. In addition, the scientists made a mixture of
hemoglobin from normal and sickle-cell individuals and placed that in the
tube. The resulting scans are distributions of concentrations along the tube,
and are made relative to the zero, no movement starting location. If a peak
is to the left of the reference no-movement position, then negative move
ment has occurred; if to the right, then positive movement has occurred.
Using electrophoresis in this way, Pauling and his colleagues showed
that hemoglobin from patients with sickle-cell anemia is different than
that of normal individuals. The evidence that they are different pro
teins includes the differential movement at pH 7.0. This made sickle-
cell anemia the first disease in which an abnormality in a protein was
known to be the cause. The scanning diagrams of hemoglobin demon
strate that normal and sickle-cell anemia hemoglobins are homogenous
substances (i.e., one protein) because only one peak occurs. The peak
for normal individuals was the same, whether they were of Caucasian
or African descent.
From the scanning diagrams, the scientists found that at pH 6.9 the
peak of normal hemoglobin distribution was below the line of no move
ment and the peak of sickle-cell anemia hemoglobin was above the line,
both as expected. For individuals that had the mild form of sickling, two
peaks were observed, as found also with a mixture of the sickle-cell ane
mia and normal hemoglobin, with one peak above and one below the line
of no movement.
However, the amount of normal hemoglobin in individuals with the
mild form of sickling was greater than in the artificial mixture. The ratio
of normal to sickle-cell hemoglobin in people with mild sickling is about
sixty to forty. Pauling and his colleagues concluded that individuals with
mild sickling have both types of hemoglobin and are heterozygous for the
gene that produces hemoglobin. Sickle-cell anemia and normal individu
als are both homozygous.
In the late 1950s Vernon Ingram began the process of trying to deter
mine the exact nature of the differences between normal and sickle-cell
disease hemoglobins. He predicted that the sickle-cell hemoglobin had
one less carboxylic acid group in its protein structure, which would ac
count for the difference in charge observed by Paulings group.
Ingram denatured purified hemoglobin. He then digested it with
trypsin, an enzyme that breaks down proteins, predominantly wherever
the amino acids lysine and arginine are found, except when either is fol
lowed by proline. The digestion produced shorter peptide chains that
were reproducible, because each type of hemoglobin would always be
broken into chains of particular amino acid composition when exposed
to trypsin. If there were differences between the sets of chains, then those
proteins could be identified as having different amino acid sequences.
Ingram performed both electrophoresis and chromatography on
these samples to separate the peptide chains in two dimensions. Peptides
are separated by both their charges in an electric current and their mobil
ity in a solvent gradient. Small peptides move faster with the leading edge
of the solvent. The proteins are then dyed, and the resulting image is the
peptide fingerprint of a protein. Ingram carefully traced the blotches of
peptides from the filter paper and numbered equivalent pairs.
The two hemoglobin fingerprints could be almost completely super
imposed on each other. One peptide (labelled #4) was in different posi
tions in normal and sickle-cell hemoglobin. It appears along the set of
peptides that are slightly positively charged. The altered peptide has a
stronger positive charge in sickle-cell hemoglobin relative to normal he
moglobin, which caused it to migrate toward the negatively charged elec
trode during electrophoresis. Superimposition of other peptides indicates
that the amino acid sequences were the same.
Ingram isolated peptide #4 from the two hemoglobins. He broke it
down into smaller chains and performed his two dimensional fingerprint
ing technique on those peptides. He knew the amino acids that were
present in these peptides, based on other analyses, and that there was
more glutamic acid in the normal hemoglobin chain and more valine in
the sickle-cell hemoglobin chain. In determining the amino acid chain for
this peptide, recall that trypsin cuts proteins wherever it finds a lysine or
an arginine. Because arginine was not found in this peptide, lysine must
be at the end of each chain. Using empirical data and knowledge of the
structures and charges of each amino acid, Ingram deduced the peptide
structure of each spot on the fingerprints.
The two longest non-overlapping sequences for either peptide chain
are four and five amino acids in length, making the minimum chain
length nine. Taking into account the overlap in sequences that he found,
Ingram lined up the sequences as in Table 2.
Ingram showed that in this particular peptide chain, the seventh posi
tion was altered, such that in sickle-cell hemoglobin a valine was substi
tuted for a glutamic acid. Glutamic acid has a carboxylic acid on its side
chain, which would make it more negatively charged than the valine, or
in the case of the full nine amino acid sequence, the negative charge on
the glutamic acid balances positive charges in the other amino acids to
make it more neutral than the peptide from sickle-cell hemoglobin.
Table 2 Amino acid sequence alignment for the peptide hemoglobin

chain #4, reconstructed from peptide fragments. E - Glutamic acid;
H - histidine; K - lysine; L - leucine; P - proline; T - threonine;
V - valine.
type of hemoglobin peptide chain sequences
H V
H V L L
V L L
sickle-cell T P V
T P V E
T P V E K
E K
reconstructed sickle-cell peptide H V L L T P V E K
H V L
H V L L
normal T P E
T P E E K
E K
reconstructed normal peptide H V L L T P E E K
Source: From Ingram, 1957, Figure 2.
Ingram correctly deduced that the sickle-cell peptide had one less car
boxylic acid group than normal hemoglobin. Scientists later determined
that Ingram misinterpreted the fingerprintthe H-V-L-L sequences
turned out to be V-H-L. Scientists now know that the amino acid substi
tution occurs at the sixth position, not the seventh; the corrected normal
sequence is V-H-L-T-P-E-E-K, and the corrected sickle-cell sequence is
V-H-L-T-P-V-E-K. A point mutation in the DNA sequence of individu
als with sickle-cell anemia leads to the altered hemoglobin. Sickle-cell ane
mia became the first genetic disorder whose molecular basis was known.
Recall that Pauling and his colleagues were aware that sickling occurred
primarily when oxygen content is low. Perutz and Mitchison (1950) ex
amined red blood cells from sickle-cell disease and normal patients. When
hemoglobin was deoxygenated, they found that sickled cells exhibited
physical properties similar to crystals, but normal cells did not. The re
searchers concluded that sickle-cell hemoglobin crystallizes in the absence
of oxygen. The scientists further went on to test the solubility of oxygen
ated and deoxygenated hemoglobins in solutions of varying ionic concen
trations (Table 3). The scientists determined the maximum concentration
of each hemoglobin in each condition that could be dissolved in solution.
The point mutation that produces sickle-cell hemoglobin creates a
protein with different physical properties from normal hemoglobin. When
saturated with oxygen, it behaves like normal hemoglobin, having the
same solubility. But when it has lost some or all of its oxygen, as it does
in venous blood, it begins to precipitate and crystallize and its solubility
declines. At any ion concentration, oxygenated hemoglobin has higher
solubility than deoxygenated hemoglobin. At ionic strength of 5.0, the
solubility of either type of oxygenated hemoglobin is 6.3 grams per liter,
Table 3 Solubility (g/L) of normal and sickle-cell hemoglobin at

different pHs. de-O2 - deoxygenated; O2 - oxygenated forms of
hemoglobin.
ionic strength normal hemoglobin sickle-cell hemoglobin
de-O2 O2 de-O2 O2
4.5 no max found no max found 0.3 no max found
5 1.6 6.3 0.08 6.3
5.5 0.1 0.4 not soluble 0.4
Source: Data from Perutz and Mitchison, 1950, Fig. 3.
whereas that of deoxygenated normal hemoglobin is 1.6 g/L and that of

deoxygenated sickle-cell is only 0.08 g/L, about 1% of oxygenated hemo
globin. The precipitation causes the concentration of dissolved solutes to
decrease inside the cell, which causes water to diffuse out of the cell, col
lapsing the cell and causing anemia. Sickle-shaped cells then block small
blood vessels, causing less blood to reach parts of the body, which can
damage entire tissue beds.
Recall that about 8% of the African American population in the
1940s exhibited the mild form of sickling and that 2.5% had sickle-cell
anemia. In a random sample of 1,000 African Americans from that time,
25 homozygotes and 80 heterozygotes would be expected to have sickle-
cell anemia and mild sickling, respectively. The frequency of the mutated
allele can be determined by counting up the expected number of sickle-
cell hemoglobin alleles in this sample, which would be 50 (two copies for
each of 25 homozygotes) plus 80 (one for each heterozygote), and then
dividing that by 2,000 (the total number of alleles in the sample of 1,000
people). The resulting allele frequency is 130/2000 = 0.065. Currently, it
is estimated that the frequency among African Americans is about 0.044,
so it has declined since the 1940s. In other populations of blacks, the
allele frequency is much higher; frequencies of heterozygotes with mild
sickling have been reported to be as high as 40% in some African popula
tions. In times and places without adequate medical treatment, individu
als with sickle-cell disease could expect to die by age 14.
If homozygotes do not live long enough to reproduce, it seems
strange that the mutated hemoglobin gene persists in human popula
tions. Evolution should eliminate harmful mutations. However, because
heterozygotes do not suffer greatly, the allele persists in the population.
Individuals with the mild form of sickling are carriers of the mutated al
lele, and two heterozygotes that mate have a 25% chance of giving birth
to an individual with sickle-cell anemia and a 50% chance of giving birth
to another heterozygote.
In the 1950s, Anthony Allison suggested that the answer to the ques
tion of variable frequencies was selective advantage in some environ
ments. In Africa he noted that incidence of the sickle-cell trait was higher
in regions where malaria was prevalent than in other regions. Allison con
trolled for the possible development of immunity by studying children
Table 4 Incidence of the malaria parasite in blood of normal and

sickle-cell children from a small community in Uganda, Africa,
andadult males dosed with the malaria parasite.
A hemoglobin genotype percent with malaria percent with high
in children parasite present parasite density
normal (homozygous normal) 45.7 66.0
mild sickling (heterozygous) 27.9 33.3
B hemoglobin genotype percent with malaria percent with high
in adult males parasite present parasite density
normal (homozygous normal) 93.3 40.0
mild sickling (heterozygous) 13.3 0
Source: Data from Allison, 1954, Table 1 and text.
under the age of 5 in a single Ugandan community where malaria was

prevalent (Table 4A). In addition, Allison determined the densities of
malaria parasites and the severity of the malaria infections.
To further investigate, Allison chose 15 adult male volunteers with the
sickling trait and 15 normal adult male volunteers, all of whom had been
away from malarious regions for at least 18 months. These volunteers
were infected with malariaan ethically questionable procedure. Every
2 days for 40 days, blood of the 30 males was checked for the malaria
parasite (Table 4B). At the end of the 40 days, anti-malarial drugs were
used to stop the infection.
The data show that a lower percentage of children with the sickle-
cell trait than normal children were infected with the malaria parasite,
and of the infected individuals, fewer sickle-cell children had severe
infections. Even though the men in the second experiment came from
a malarious region, had been exposed to malaria in their pasts, and
probably carried some immunity, the incidence of malaria during the
trial was seven times higher in normal individuals than in sickle-cell
individuals. Even if malaria takes hold in sickling individuals, the symp
toms are less severe and last for shorter periods of time. Frequencies of
sickle-cell hemoglobin allele correlate with the prevalence of malaria.
The malaria parasites complete part of their life cycle within red blood
cells; their activity uses up oxygen in the cells, causing cells of hetero
zygotes to sickle. These cells are removed from the body by the spleen,
eliminating the parasite at the same time. Thus there would be selection
for heterozygous individuals in malarious regions.
Multiple Mutations may Cause a Single Disease

Sickle-cell disease is caused by a single point mutation at a specific genetic
locus. Other diseases are much more complex, and may have multiple
causes and multiple mutations, only some of which may be responsible
for the manifestation in any one patient. A complex group of diseases,
with both genetic and environmental causes, is motor neuron disease
(MND), and this group of diseases has variable effects.
Amyotrophic lateral sclerosis (ALS; also known as Lou Gehrig disease)
is incurable and is the most common form of MND with about 10% of
cases inherited, known as familial ALS. ALS is a progressive neurodegen
erative disease that affects nerve cells in the brain and spinal cord. Motor
neurons degenerate, causing scarring, or sclerosis, in the spinal cord and
brain, leading to wasting away of muscles controlled by the affected
nerves. Patients diagnosed with ALS have a life expectancy after diagnosis
of 2 to 5 years. The incidence of the disease is very low, about 0.002% of
humans. About 20% of familial ALS cases are caused by a particular mu
tation, but the causative gene for most cases of familial ALS is unknown.
Kwiatkowski and his colleagues studied several families where ALS
was common and found one family from islands off the west coast of
Africa that had four members develop ALS in their upper and then lower
extremities. The maternal grandparents in this family were first cousins,
and the population on the island from which they were from was rela
tively small; so the researchers suspected that the inheritance pattern of
the ALS trait was recessive. The researchers conducted a genetic study to
determine areas of the genomes in family members that had lost wild type
alleles, either through mutation or chromosome loss. They did this for
a variety of genetic loci, including several suspected of being associated
with ALS. They found an area of chromosome 16 associated with ALS, as
well as several other regions of the genome, to have lost wild type alleles.
A large area on chromosome 16 contained approximately 56 genes that
were candidates in causing ALS.
The researchers sequenced this 4 million base pair region of DNA. In
the family members with ALS, the researchers discovered multiple mu
tations in the fused in sarcoma/translated in liposarcoma (FUS/TLS)
gene. The FUS/TLS protein regulates how gene messages are created,
modified, and transported in order to make proteins. It happens to link

with another protein, TDP-43, which is deposited in motor neurons in
90% of all people with MND. This alone suggests a possible role in caus
ing familial ALS.
In the FUS/TLS gene, exon 5 has two mutations, one that results in
the insertion of two glycines (called insGG) and one that leads to two
glycines being deleted (delGG). In exon 6 there is a mutation in the
genetic code that causes a cysteine in position 244 to be replaced by argi
nine. The amino acids in positions 514 to 526 from exon 15 of the FUS/
TLS protein have 10 known mutations, just in that region.
In one mutation, a guanine is replaced by a cytosine at position 1551
of the DNA, causing the amino acid glutamine to replace histidine in the
resulting protein. All individuals in the study family had this mutation;
unaffected individuals were heterozygous for this allele. Of the family
members who were homozygous with the mutated allele, four were af
fected and three were below the age at which ALS typically strikes. (They
might develop ALS later in life.) In 1,446 control samples from individu
als living in North America, Kwiatkowski and his colleagues never found
this particular mutation.
The researchers next sequenced and screened exons for the FUS/TLS
gene in seventeen other families suffering from familial ALS. The sci
entists discovered thirteen mutations that were co-inherited with ALS,
meaning that most individuals with the mutations were stricken with
ALS, and no individuals without the mutation developed the disease.
These 13 mutations were not found in the 1,446 North American control
subjects that did not have ALS.
The mutations that cause familial ALS were especially common in
exon 15 of the FUS/TLS gene. Of the 20% of all ALS cases that are fa
milial, about 20% are caused by a mutation in a one gene that codes for
an enzyme, and 5% are caused by mutations in the gene that produces the
TDP-43 protein. Kwiatkowski and his colleagues estimated the thirteen
mutations in the FUS/TLS gene account for another 5% of familial ALS
cases. That means that up to 70% of familial ALS cases still have un
known genetic causes!
Normal FUS/TLS protein is found in motor neurons (which con
vey electrical impulses from the nervous system to a muscle or gland),
binds to RNA, and regulates how messenger RNA is created, modified,

and transported out of the nucleus. Some of these functions overlap with
functions of TDP-43, implying that loss of these functions can lead to
familial ALS. As might be expected, FUS/TLS is normally located in the
nucleus.
To determine how the mutations affect the function of FUS/TLS,
Kwiatkowski and his colleagues studied the cellular distribution of the
normal and mutated proteins. The investigators produced FUS/TLS-
specific antibodies and covalently linked green dye to the antibodies. They
also produced antibodies to several other proteins linked to dyes of other
colors. When brain and spinal cord cells from deceased control and ALS
patients are mixed with the antibody, the antibodies bind to their spe
cific proteins. The scientists detected the presence and cellular location of
FUS/TLS and the other proteins using fluorescent microscopy.
The researchers examined neurons exposed to the different fluorescent
antibodies. One antibody was specific to a protein known to be usually
found in the nucleus, which was used as a marker to determine the loca
tion of the nucleus. The procedure revealed staining of FUS/TLS in the
nuclei of normal and ALS individuals, but the mutated protein in the
ALS individual also occurred in the cytoplasm. Although Kwiatkowski
and his team found that some mutations led to high abundance of the
protein outside the nucleus relative to control individuals, they did not
find that was true for all familial ALS cases.
Using an antibody for another protein, ubiquitin, they found that
ubiquitin localized in ALS patients neuronal nuclei but not in controls.
Ubiquitin is a protein that covalently binds to other proteins, which it
does to tag the proteins so that degradation enzymes can find them.
Proteins that dont function properly are often tagged for elimination by
ubiquitin. Antibodies to ubiquitin are used to identify abnormal accumu
lations of protein inside cells, and ubiquitin was found everywhere in the
normal cell, but much more likely to be found in nuclei with FUS/TLS
in ALS patients. The mutations then led to altered proteins, some that
localized in the wrong place and some that may have folded incorrectly, as
evidenced by the binding of ubiquitin to the mutated protein. Ubiquitin
in normal individuals does not localize in nuclei and is rather diffusely
distributed throughout the cell. This indicates that there are no unusual
proteins being tagged by ubiquitin. Usually the FUS/TLS protein func

tions in DNA and RNA metabolism in the nucleus, but the mutation
causes the protein to be abnormally located outside of the nucleus, where
it forms large aggregates within motor neurons.
Kwiatkowski and other researchers continue to work on how mutated
proteins, such as FUS/TLS and TDP-43, actually cause MND. It turns
out that other genes that have functions involving DNA and RNA me
tabolism are associated with other MNDs. This knowledge could lead to
elucidation of mechanisms that lead to these deadly genetic diseases.
Inherited diseases of the whole individual are caused by disruptions
in normal functioning of both red blood cells and neurons. In both cases,
mutations produce altered proteins that disrupt the particular cells in
which the proteins are located. Disrupted protein functions then lead to
disrupted ability of the entire cell to function and that manifests itself in a
variety of symptoms. If cells cannot maintain homeostasis, structure, and
function or communication with other cells, an entire multicellular or
ganism may be affected to the point of death. In the next chapter, we turn
to pathogenic diseases to learn how pathogens cause disruptions in cells.
Ethical, Legal, Social Implications:

NormalityandPerfection
How is normality defined? The term normal was used to describe indi
viduals not afflicted with a genetic disease. Before we can discuss what is
normal in the human species, we must define what normal means. It may
be defined as conforming to a type or a standard, for instance, of a be
havior. A person with normal behavior conforms to a standard such that
she behaves a certain way. By not breaking any laws or codes, she exhibits
normal behavior. We may also define it as an average quality of some trait
or character. In that case, we recognize that there is variation around the
average. For instance, a person may be of average height, but in a large
population there are people of many heights.
In the early beginnings of Western thought, a species was defined as
an ideal type, and all living representatives of that species were imperfect
imitations of the ideal. Aristotle and other pre-Darwinian philosophers
and scientists took the species to be distinct and unchanging with an
essence, like the chemical elements. That would make each of us im

perfect imitations of the ideal human. Variation exists in just about every
population.
What about perfection? Perfection is defined as being without flaws.
Something that is perfect also corresponds to an ideal standard. The stan
dard used in our normal definition does not refer to an ideal standard.
For instance, a person that exhibited normal behavior in every aspect of
his life may not be perfect in the sense that he may not exhibit the ideal
standard for behavior, yet he still has not broken any laws or moral codes.
Perceived normal traits do not always align with the perceived perfect
example of that trait. The perfect height may not be the average height.
Perfection varies, depending on point of view or cultural context.
If normal is average and perfect is being better than normal or bet
ter than average without any flaws, then most people fail to be perfect.
Many people are not happy with the way we are, and believe they are
imperfect or flawed. They are too tall, too short, their nose is too big,
their hair is falling out, or their eyes are not blue. They may be unhappy
with their mental abilities. However, if everyone were perfect, then that
defines the new average, normal condition, with no variation around that
condition. Imagine a world where all the dogs were the same breedlets
say they are all Labrador Retrievers. Although this is a popular breed of
dog, should all dogs be Labs? The same may be true for humans: Most
people do not want to live in a world where everyone looked, acted, or
thought the same, even if we all agreed that those looks, actions, and
thoughts were perfect. Although this might sound beneficial because it
might reduce wars and conflict and make it easier to find a mate, there
are some potential negative consequences.
If everyone were perfect, there would be little genetic variation in the
population if, that is, the source of perfection were genetic. We humans
now have the ability to create perfection through genetic engineering,
plastic surgery, and Botox injections. Diversity serves several purposes in
human populations. Without diversity, a population may be less able to
withstand a new disease or a catastrophe, because if there is no variation
in susceptibility, the entire population may be exterminated. Our lives
as humans are enriched when exposed to a diversity of cultures, ideas,
and experiences. If everyone looked and acted alike, life might be pretty
dull and boring. In some countries, like the United States, people value
individuality of thought and expression, and this could be lost in a world
where everyone thought and looked alike.
These examples illustrate the extreme. With more than 7 billion peo
ple on the planet, it is safe to say that we will remain diverse in looks and
thought for many years to come. As long as that diversity of thought
remains, then what it means to be perfect will vary, and humanity may
never converge on a single way of thinking and looking.
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CHAPTER 2
Pathogens Affect Cells

andOrganisms
Much can be learned about how cells function by learning how vari
ous environmental factors disrupt cell function and homeostasis. In this
chapter, how some pathogens cause disease by disrupting cell functions
will be examined. Pathogens can have similar effects to those of genetic
diseases, in that they can disrupt cell function, which disrupts function
at the organismal level, depending upon what specific cells pathogens are
attacking. One pathogen that affects humans and another one that affects
plants will be used to illustrate general effects of pathogens.
Lyme Disease Is Caused by a Vector-Borne Bacterium

Lyme disease is an emerging disease, an infectious disease that is new in a
population or rapidly increasing in incidence or geographic range because
it has sprung up relatively recently. It is caused by the bacterium Borrelia
burgdorferi and is a vector-borne disease transmitted by ticks to mam
mals, including humans. Vector-borne diseases are diseases in which the
pathogen is transmitted to an individual by some other agent. The tick
is the vector, and the mammal is the host. Tick saliva and bacteria both
enter the host, and both may contain substances that disrupt the immune
response at the site of the bite. The bacterium does not cause symptoms
in some mammals in which they live; these mammals are reservoirsthey
harbor the pathogen, but it does not affect them.
In humans, Lyme disease causes many problems. There are dozens of
known symptoms, including fatigue, loss of muscle tone in the face, severe
headaches and neck stiffness due to meningitis, joint pain, heart palpi
tations, and dizziness. In later stages, patients experience joint swelling
and arthritis, numbness in the extremities, and cognitive problems. The

bacteria spread to numerous organs as would be expected if it causes this
great range of symptoms.
When the bacteria first invade a host during a tick bite, a character
istic bullseye targetshaped rash develops around the bite. Both genetic
and molecular changes occur in B. burgdorferi as it moves from the vector
to the host. These changes are important for successful host invasion,
because the environment inside of the ectothermic tick is much differ
ent than the environment inside of an endothermic mammal. Learning
about how cells invade other cells will help illustrate several of the themes
of the cell as the fundamental unit of life.
Dorothee Grimm and her colleagues studied proteins that appear on
the surface of B. burgdorferi to determine their role in transmission and
survival in either the vector or the host. In particular, they studied an
outer surface protein, OSP-C. Other studies had suggested that OSP-A
was critical for B. burgdorferi to adhere to the lining of the ticks gut,
where it remains dormant until the tick takes a blood meal. Ticks attach
and feed for several days, giving time for the bacteria to move from the
gut to the salivary glands, where they are then transmitted to the host.
There is a switch from expression of OSP-A to OSP-C during tick feed
ing. Ingestion of mammalian blood by the tick is thought to trigger this
change in the bacteria, and reduced expression of OSP-A would dislodge
the bacteria from the lining of the ticks gut.
Grimm and her colleagues investigated the role of OSP-C in B.burg-
dorferi in both the tick vector and mammalian hosts. They created a
mutant B. burgdorferi strain that lacked the ability to produce OSP-C
(OSP-C negative). From that they created another mutant that had the
OSP-C gene reinserted in another location in the genome. This led to
three strains for their experiments: one that naturally produced OSP-C,
one that did not, and one that did due to reinsertion of the gene. The
scientists confirmed that the OSP-C negative genome was the same as
the wild type except for loss of the gene that produces OSP-C, and they
confirmed that the mutant bacteria no longer produced OSP-C. They
further confirmed that the OSP-C reinserted strain did produce OSP-C.
Each strain of B. burgdorferi could successfully colonize and replicate in
tick guts as well as migrate to the salivary glands of feeding ticks.
Pathogens Affect Cells andOrganisms 17
To determine whether mutant bacteria could infect mammals, Grimm

and her colleagues injected mice or exposed mice to ticks infected with
one of the three strains of B. burgdorferi. The scientists then tested for
antibodies to B. burgdorferi and presence of the bacteria in mouse tissue;
presence of antibodies would confirm that the mice had been infected
with the bacterium. Wild type B. burgdorferi led to a production of anti
bodies in 77.8% of mice after being injected and 83.3% of mice that were
subjected to a tick bite. The scientists found B. burgdorferi in the tissues
of 44.4% and 83.3% of mice after injection or tick bite, respectively. Un
infected ticks that were allowed to feed on mice that were infected with
wild type B. burgdorferi picked up the bacteria 17.6% or 92.8% of the
time, depending upon whether the original infection route was injection
or tick bite, respectively.
Mice infected with OSP-C negative B. burgdorferi never produced
any antibodies, never produced any B. burgdorferi in their tissues, and
were not able to reinfect new ticks, whether infected by injection or tick
bite. However, OSP-C reinserted B. burgdorferi led to a production of
antibodies in 66.7% of mice after being injected and 75% of mice that
were subjected to a tick bite. Those same mice that produced antibodies
all had B. burgdorferi in their tissues. Uninfected ticks that were allowed
to feed on mice that were infected with wild type B. burgdorferi picked up
the bacteria 18.5% or 56.7% of the time, depending upon whether the
original infection route was injection or tick bite, respectively.
Grimm and her colleagues demonstrated that the OSP-C negative
mutant cannot infect mice but could survive in ticks. Mice infected with
the wild type strain produced antibodies to B. burgdorferi antigens, and
they found the bacteria in mouse tissues. Mice infected with the OSP-C
negative strain did not produce antibodies and did not contain bacteria.
The OSP-C reinserted strain was able to infect mice in a manner similar
to the wild type strain. Ticks that fed on mice infected with the wild
type or the OSP-C reinserted strain of B. burgdorferi often acquired the
bacteria.
OSP-C production is turned on in a feeding tick prior to movement
to the salivary gland of the tick, and Grimm and her colleagues showed
that OSP-C is required to infect mice. The researchers concluded that
OSP-C protein is an essential component for early mammalian infection.
Although the exact role of OSP-C in the infection cycle is still unknown,
it may be required for this pathogenic bacterial cell to evade the immune
system during the early phase of infection and spread through the blood
stream of the host.
The immune system of mammals consists of millions of cells of several
different types that play various roles in protecting the individual from
pathogens and other foreign substances. These cells move about the body,
protecting the individual. Their development, movement, and adhesion
to foreign invaders are determined by chemical signaling. Molecules used
in chemical signaling in the immune and other systems are cytokines,
small proteins, peptides, or glycoproteins released by cells involved in cell-
cell interaction and communication.
Chemokines are small protein cytokines that have receptors on spe
cific cells. When the chemokine binds to the receptor, it elicits a response
from the cell. For instance, leucocytes, or white blood cells, may respond
to a gradient of chemokine molecules by moving toward areas of higher
concentration. The release of the chemokine from tissues during bacte
rial infection or inflammation attracts leucocytes to the area of infection,
where they perform their function of attacking the invaders. Neutrophils
and monocytes are phagocytic leucocytes involved in the inflammatory
response. Phagocytes are white blood cells that ingest microbes, other
cells, and foreign particles.
In an experiment on early infection of mammalian hosts, Xu and his
colleagues investigated the ability of B. burgdorferi to evade early detec
tion, and thus establish a chronic infection. Recall the bullseye target
shaped swollen area commonly associated with the early phase of Lyme
disease and B. burgdorferi infection. This is an inflammatory response,
meaning the immune system is responding to the infection; lymphocytes
infiltrate infected tissues after being attracted to the tissues by chemo
kine signals. Several types of white blood cells are attracted to the site of
infection, yet there is a noted lack of neutrophils in B. burgdorferi infec
tions. This is unusual because neutrophils are the most common type of
lymphocyte and are often the first to arrive at sites of bacterial infection.
Clearly there is an inflammatory process in B. burgdorferi infection, yet
the immune response does not effectively clear the pathogen from the
body of the host as it should during inflammation. Xu and colleagues
performed a study to determine if this lack of neutrophil attraction was a

factor in causing Lyme disease.
To test how the lack of neutrophils affects successful invasion of
B.burgdorferi, Xu and his colleagues devised an experiment that artificially
increased neutrophil recruitment and activation during early B. burgdor-
feri infection. They created a bacterial plasmid that produced a mouse
chemokine known to attract neutrophils. The researchers sequenced the
construct to verify that what they had intended to construct was actually
constructed. Ultimately, they created plasmids that could replicate inside
both Escherichia coli bacteria and B. burgdorferi.
Xu and his colleagues grew B. burgdorferi that contained the new plas
mid. They verified that the new strain produced the chemokine in labo
ratory cultures and found that this genetically engineered B. burgdorferi
produced KC and released it into the surrounding environment. Xu and
his colleagues found that over 97% of the KC found was outside of the
cells. KC was not detected in a non-engineered strain. This allowed the
researchers to determine how B. burgdorferi invasion would proceed when
neutrophils were attracted to the infection site.
Xu and his colleagues centrifuged a culture of genetically engineered
B. burgdorferi cells, took the supernatant, and injected it into mice. Su
pernatant from cells centrifuged from E. coli cultures genetically engi
neered to express KC was used as a positive control, and supernatant from
cells centrifuged from non-genetically engineered B. burgdorferi was used
as a negative control. (Neutrophils should not be attracted to the site, as
usual.) After receiving two injections, mice were sacrificed. The research
ers collected and stained leucocytes from each mouse, and they counted
and identified the first 200 leucocytes that they observed in the extracel
lular fluid (Figure 1).
Mice were also injected with two side-by-side inoculations. One
injection contained a high dosage of B. burgdorferi with the engineered
KC plasmid, and the other injection contained a high dosage of non-
engineered B. burgdorferi. Mice were sacrificed 6, 16, 24, or 72 hours
later, and the tissues near the injection site were collected for determina
tion of cell types present. The researchers observed that non-engineered
B. burgdorferi injection sites experienced an inflammatory response, as
expected. Macrophages (a type of phagocyte), neutrophils, and basophils
percent of 200 cells that were neutrophils or monocytes

100
= neutrophils
= monocytes
80
60
40
20
0
Bb ( control) Ec w/KC (+ control) Bb w/KC
Figure 1 KC activity of cell culture supernatants. The first
200leucocytes observed in each sample were identified as either
neutrophils or monocytes. The percentage shown is the overall
percentage for all mice sampled. Bb - B. burgdorferi; Ec - E. coli;
KC- chemokine.
Source: From Xu et al., 2007, Table 3.
were all found within the first 6 hours, but by 16 hours the latter two cell
types had disappeared and eosinophils appeared and stayed for 72 hours.
In the genetically engineered B. burgdorferi injection sites, the researchers
found large numbers of neutrophils within the first 6 hours whose num
bers peaked at 16 hours and were still present at 24 hours. By 72 hours
the neutrophils had disappeared.
Other mice that received only one high dosage injection of either KC-
producing B. burgdorferi or normal B. burgdorferi were killed and exam
ined 30 days post-injection for B. burgdorferi in the heart, joints, and skin
(Table 5A). Finally, one last set of mice was given one injection each, but
the dose of B. burgdorferi cells varied. Xu and his colleagues performed
this test to determine the lowest dose that infected mice. After 30 days,
mice were killed and inspected for infection in the heart, joints, and skin,
as with the other set of mice (see Table 5B).
Table 5 A, Percentages of tissues from ten mice injected with either

non-engineered or genetically engineered B. burgdorferi that showed
active bacterial infections 30 days after injection. B, Percentages of
tissues from six mice injected with a particular dose of either non-
engineered or genetically engineered B. burgdorferi that showed active
bacterial infections 30 days after injection.
A heart joint skin
non-engineered B. burgdorferi 100 100 100
KC chemokine B. burgdorferi 30 30 60
B heart joint skin
100,000 non-engineered cells 100 100 100
100 non-engineered cells 83.3 83.3 83.3
10 non-engineered cells 0 0 0
10,000,000 KC chemokine cells 33.3 16.7 66.7
1,000,000 KC chemokine cells 0 0 16.7
100,000 KC chemokine cells 0 0 0
Source: From Xu et al., 2007, Tables 4 and 5.
By artificially attracting neutrophils to the site of a B. burgdorferi

infection, Xu and his colleagues could test the effects of neutrophils
on successful invasion of the pathogenic bacteria. Neutrophils are part
of the defense against infection, and without them the inflammation
process may not successfully rid the body of foreign invaders. Only
macrophages and neutrophils can directly kill pathogenic bacteria. The
scientists showed that KC attracted neutrophils, and neutrophil recruit
ment enhanced the hosts ability to eliminate the bacteria; these leuco
cytes could directly kill the bacterial cells. Xu and colleagues found that
all mice injected with non-engineered B.burgdorferi were infected with
bacteria in all tissues sampled, but a smaller percentage injected with KC
chemokine B. burgdorferi were infected, with the infection not consis
tently present in all tissues (Table 5A).
The researchers also found that dose mattered. It took high doses of
KC B. burgdorferi to establish a successful infection, whereas it took only
a small dose of normal B. burgdorferi to successfully invade and estab
lish populations in mice. The researchers did not even test doses of KC
engineered B. burgdorferi below 103 cells, because no mice below 106 cells
showed any evidence of successful invasion. By testing the mice in order
from high to low dose, they were able to reduce the total number of mice
tested because it would be unlikely that any mouse given a dose lower
than 103 would react.
It is unknown how B. burgdorferi normally evades the hosts immune
system, but the mechanism results in lower attraction of neutrophils to
the site of infection. Tick saliva has been shown to reduce neutrophil
function and attraction to a tick bite. Perhaps B. burgdorferi takes advan
tage of this mechanism that ticks use to evade the immune system. There
is a normal inflammatory response that causes neutrophils to be recruited
early on in the infection, but neutrophils disappeared quickly in mice
infected with normal B. burgdorferi, allowing the bacteria to establish
within the host. The high numbers of macrophages, which kill bacteria,
suggest that the bacterial infection would be eliminated, but for unknown
reasons, the macrophages were not effective against B. burgdorferi.
After the initial infective population of B. burgdorferi is established,
the bacteria migrate to joints, the heart, the nervous system, and other
tissues. Once they establish populations in other tissues, Lyme disease
manifests itself in many ways. Researchers have found that these bacteria
evade further immune responses by decreasing expression of OSPs. These
OSPs may be targeted by antibodies, and antibodies cannot recognize
invaders without the protein present. The bacteria can hide in the extra
cellular matrix and tissues where the immune system may not be able to
respond effectively. Disruption of the immune system by the pathogen
causes chronic inflammation even after the pathogen is gone from the
system, and may be an example of pathogen-induced autoimmunity.
Ethical, Legal, Social Implications: There are

SeveralIssues with Using Animals in Research
In humanitys quest for knowledge and cures for diseases, we often used
animals as subjects of our experiments. Basic knowledge is gained by per
forming experiments on animalsknowledge of the animals, but also
of ourselves. Scientists use animals as model organisms, upon which we
experiment to learn more about diseases that afflict us. Much information
has and can be gained from performing research on animals. Researchers

used mice to study Lyme disease in this chapter. To protect environmental
and human health, many governments require testing of new chemicals
on animals to determine if they are safe for use.
Strong feelings exist on either side of the animal research debate.
Supporters argue that medical and scientific knowledge will not advance
without research of all kinds, including research on animals. Those who
argue against animal research make the claim that we have no moral right
to exploit other creatures for our gain, and we will not advance as a soci
ety while our fellow creatures suffer for our benefit. Some opponents of
animal research have committed illegal trespass to break into laboratories
and set caged animals free. In truth, much of the research on animals
is done for their benefit (e.g., conservation and ecological research) and
causes no harm (e.g., behavioral observations, population counts). Con
sidering the range of experiments, from nonintrusive behavioral observa
tions to painful medical procedures, where should we draw the line on
what should be ethically acceptable?
Those who argue for animal experimentation generally put human
health and safety as the top priority, above that of other animals. When
there is the probability that experiments on animals will lead to a medical
breakthrough, then animal suffering is justified. All federally funded animal
experiments in the United States must follow a set of guidelines to minimize
animal suffering and provide them with a reasonably high quality of life.
In addition, it is the law in some countries that pharmaceuticals and pesti
cides be tested on animals prior to approval for use. In these cases, only live
animals will be adequate subjects. If not tested prior to release, pesticides
could cause more harm and kill many more animals than were used in the
testing. Because all animals share an evolutionary history, testing on closely
related organisms provides useful information on how a chemical will act in
humans. This is far better than testing the chemical on humans; one need
only consider the ethical ramifications of exposing men to malaria.
Those who oppose animal experimentation value humans and other
animals more or less equally. Animals have rights as individuals, and
should not be used to advance human causes. To expose mice to Lyme
disease, as Xu and colleagues did, is unacceptable, no matter what might
be gained from it. In addition, the sheer numbers of laboratory animals,
ranging in the millions around the world, is much more than is needed
and is unacceptable from the animal rights advocates perspective. The
more animals held in captivity, the greater the chance that abuse, mis
treatment, or cruelty will occur. Advances in growing cell and tissue cul
tures will allow experiments to be done without the use of animals. Even
if we argue that mice are fairly closely related to humans (and they are
in the sense that both are mammals), the results from studies such as the
Lyme disease experiments may not be a useful guide to what is happening
when B. burgdorferi infects a human. If humans used only animals that
were very closely related to them (such as, chimpanzees), they are being
even more unethical, because chimpanzees are intelligent and sentient be
ings. If the humans dosed with malaria were not experimented on unless
they gave their consent and humans and other animals have equal value,
then humans cannot justify experimenting on an animal that is not able
to consent to the procedure.
Fungi can be Pathogens of Plants

A bewildering diversity of pathogenic organisms exists, and the mecha
nisms they use to disrupt cell structure and function are almost equally
diverse. Borrelia burgdorferi uses several mechanisms to successfully in
vade mammalian hosts. Although many pathogens are bacteria, many
are not. Pathogens of plants include fungi, bacteria, viruses, protozoa,
and nematodes. An example of an interaction between two organismsa
plant host and its fungal pathogenwill be examined to learn how cells
are disrupted in plants by at least one pathogen and how cells in the
pathogen change to successfully invade the host.
Many fungi pathogenic to plants belong to one phylum within the
Kingdom Fungi: the ascomycetes. Phylum is the term used to denote the
primary division of a kingdom, ranked above class in size. Most ascomy
cetes fungi, or sac fungi, reproduce sexually via the production of spores.
Sac fungi include beneficial fungi, such as morels, truffles, and brewers
yeast, and several economically injurious plant pathogens. Magnaporthe
grisea, or rice blast fungus, causes lesions on rice leaves. A blast is a term
used to describe a syndrome of plant diseases where afflicted leaves sud
denly die. When conditions are right, lesions grow rapidly to completely
cover leaves. Some individuals are resistant to infection; those plants de

velop small brown specks, but the specks do not expand.
Rice blast fungus has to undergo several growth stages leading to the
production of infection cells. These dome-shaped cells have a complex
cell wall structure and must physically penetrate the host cell. Richard
Howard and his colleagues studied the penetration process. There are two
major hypotheses for how penetration occurs. One deals with enzymatic
breakdown of the plant cell wall, and the other deals with mechanical
force breaking through the cell wall. Rice blast fungus infection cells ad
here strongly to the surface of the host plant, producing thin outgrowths
that penetrate. The chemical, melanin, accumulates in the cell walls of
infection cells, which creates a highly concentrated solution between the
cell wall and the cell membrane and causes water to build up in this
space. The increased water volume increases pressure. Howard and his
colleagues tested the hypothesis that this increased pressure was respon
sible for penetration.
The scientists grew the fungus on artificial substrates in the labora
tory. The artificial surfaces were six different types of a plastic polymer,
which each differed in density and hardness. Howard and his colleagues
estimated the pore size of infection cell walls with melanin. Then they
blocked melanin synthesis using a chemical that blocks melanin synthesis
in fungi, and measured the pore size without melanin. The procedure
to estimate pore size involved exposing infection cells to concentrated
solutions of polyethylene glycol (PEG) polymers, each of different mean
molecular weight and size. The researchers screened the cells for either
plasmolysis or cell collapse (Table 6). Plasmolysis is the bursting of a cell
due to influx of a large abundance of molecules.
Table 6 Response of rice blast infection cells when exposed to

concentrated solutions of PEG polymers of different mean molecular
weights.
PEG molecular weight cells with melanin cells without melanin
200 (<1 nm pore size) >90% collapse >90% burst
400 (1 nm pore size) >90% collapse 90% burst
600 (2 nm pore size) >90% collapse 90% collapsed
Source: Data from Howard et al., 1991, text.
Cells burst (plasmolyze) when exposed to PEG molecules small enough

to enter the pores, because a high concentration outside the cell would
cause rapid diffusion of many PEG molecules and bursting of the cell. Cells
would collapse when exposed to a high concentration of PEG molecules
too large to enter the pores, because that would cause water to diffuse out,
essentially emptying the cell. The diameter of the pore can be estimated
by the size of the smallest molecule that did not cause plasmolysis. If a cell
collapses, then water had left the cell, and the pore size was smaller than the
smallest PEG that caused collapse; but in the case of cells without melanin,
larger pore size than the largest PEG caused plasmolysis.
Howard and colleagues found that 90% of cells without melanin
plasmolyzed when exposed to PEG molecules with an average molecular
weight of 400, whereas 90% collapsed when exposed to 600 molecular
weight PEG. Based on size of the PEG molecules, they estimated that
infection cells without melanin have a pore size of 1 to 2 nm. The smallest
PEG, 200, caused collapse in cells with melanin, indicating a smaller pore
size (<1 nm) than cells without melanin. Melanin is required for suc
cessful invasion by rice blast fungus; rice blast fungus mutants that cannot
produce melanin are nonpathogenic and cannot invade. Loss of melanin
production prevents the mutants and chemically-treated cells from de
veloping high internal pressure. Small pores would prevent loss of large
molecules from the cell, which would help retain water and thus develop
internal pressure needed for penetration.
The scientists next measured the pressure that builds up in the cell
over time by estimating the external osmotic pressure that induced cell
collapse in 50% of a test population of infection cells. Osmotic pressure is
the pressure exerted by water through a semi-permeable membrane sepa
rating two solutions with different solute concentrations.
To create different levels of internal pressure, they grew infection cells
in water on a substrate for different lengths of time. Then they replaced
the water with a solution of PEG known to be too large to enter the cells.
Using molecules on the outside of the cell that were too big to enter the
cell allowed researchers to estimate internal turgor pressure. By varying
the concentration of the PEG solution, they created a variety of external
osmotic pressures. The external osmotic pressure that caused 50% col
lapse was used as the estimate of internal pressure.
Once the external concentration got too high, the cells collapsed and
lost their internal pressure. At that point cells were no longer able to
penetrate the surface on which they adhered, and that told the researchers
that external was close to internal pressure. Because not all cells behave
the same when exposed to a particular environmental condition, the sci
entists used the point at which 50% of the cells collapsed.
The scientists found that internal pressure increased prior to polymer
penetration. The longer the cells incubated prior to exposure to a PEG
solution, the greater the external pressure needed to be in order to cause
a high degree of cell collapse. In other words, over time the internal pres
sure builds up in these infection cells and thus in order to cause collapse
a higher external pressure is needed.
The scientists next let infection cells attach to and attempt to penetrate
the six different polymer sheets for different lengths of time, and then 100
cells were tested for their ability to penetrate the substrate. High pressure
in infection cells allowed penetration of even the hardest substrate, but
the harder the substrate, the more incubation time was required.
Howard and his also colleagues determined the pressure that inhib
ited penetration of substrates; if penetration occurs at a particular external
pressure, this indicates that internal pressure is high enough to penetrate
the particular substrate. To test this, infection cells were grown in water
for 18 hours, after which the water was replaced with various concentra
tions of a very large PEG. When the external osmotic pressure increased,
penetration was inhibited severely even for hard surfaces.
From these clever experiments, Howard and his colleagues determined
that pressure built up in infection cells and that the longer pressure built
up, the harder the surface was that could be penetrated. The enormous
pressure was shown to allow penetration of plant cuticles. Once inside the
plant cell, the invading fungus swells and fills the cell. As the outside struc
ture grows, neighboring epidermal cells of the rice plant are penetrated by
more cells. A few days after multiple penetration events, visible symptoms
of infection can be observed. As the infection grows, the fungus disrupts
cells in the epidermis of the rice leaf, and as multiple areas grow together a
gray lesion develops where fungal spores are produced.
Individual hosts can be infected by pathogens and that pathogens have
unique structures that function to disrupt host cells and allow invasion of
the host. Host cells may no longer be able to maintain homeostasis and
normal function when invaded by pathogens, which can cause disease or
death. Pathogenic cells may communicate with host cells, causing them
to malfunction, or the pathogens can invade host cells. Either way, cell
function is often disrupted, leading to problems for the entire organism.
In the next chapter, more about parasites will be explored, in particular,
how it is that parasites can survive in multiple host species.
Bibliography
Grimm D, Tilly K, Byram R, et al.: Outer-surface protein C of the Lyme
disease spirochete: a protein induced in ticks for infection of mam
mals, Proc Nat Acad Sci 101(9):31423147, 2004.
Howard R, Ferrari MA, Roach DH, et al.: Penetration of hard substrates
by a fungus employing enormous turgor pressures, Proc Nat Acad Sci
88(24):1128111284, 1991.
Xu Q, Seemanapalli SV, Reif KE, et al.: Increasing the recruitment of
neutrophils to the site of infection dramatically attenuates Borrelia
burgdorferi infectivity, J Immunology 178(8):51095115, 2007.
CHAPTER 3
Parasites can Survive

inMore than One Host
Species
Populations of organisms interact with each other, and one common in

teraction is parasitism. In this chapter, how one single-celled organism can
affect more than one other species will be considered. Pathogenic organ
isms have been explored in earlier chapters on how parasites cause disease.
Life cycles of parasites have evolved to facilitate the survival, growth, and
dispersal, the act or process of spreading of organisms from one place to
another, of these complex organisms. The concepts of survival, growth,
and dispersal are universal to all organismsthey must all survive, grow,
and disperse in order to be successful. These concepts will be examined
in the context of ecological systems, and how other living and nonliv
ing components of the ecological system affect the parasitic organism. In
turn, the parasite affects its environment, including its host species.
One parasite that is a common cause of diarrhea in humans around
the world is Giardia lamblia (Figure 2). Often just called Giardia, this
parasite is a unicellular eukaryote with flagella, long tapering appendages
that project from many microorganisms that use them to move.
Robert Black and his colleagues with the Centers for Disease Control
and Prevention (CDC) studied an outbreak of diarrheal illness at a day
care center. An outbreak is an occurrence of disease greater than would
otherwise be expected in a particular time and place. They also studied
children in two other day care centers that did not have a recognizable di
arrheal outbreak, as well as children in the surrounding community who
did not attend any day care center but did obtain their water from the
same municipal water source. The researchers determined which children
flagella
Figure 2 Giardia lambla trophozoite.

Source: http://commons.wikimedia.org/wiki/Category:Giardia#mediaviewer/File:Giardia_muris_
trophozoite_SEM_11643.jpg. CDC/Dr. Stan ErlandsenThis media comes from the Centers
for Disease Control and Preventions Public Health Image Library (PHIL), with identification
number #11643. Public domain.
had experienced diarrhea in the past 2 months and, for a subset of each
of the three groups of children, obtained fecal samples on three separate
occasions to test for Giardia (Figure 3A).
Part of each fecal sample was preserved in 5% formalin and exam
ined microscopically for Giardia cysts, capsules, or resistant covers formed
around microorganisms in resting stages. Another part of each sample was
preserved, stained, and examined microscopically. The staining method they
used is rapid; it preserves the microscopic organisms and allows researchers
to distinguish among different amoebae and flagellates. The scientists also
interviewed the parents and collected fecal samples from any 1- to 2-year-
old siblings of children enrolled in the day care center (see Figure 3B).
Giardia was much more prevalent in the children who attended the
day care center with the diarrheal outbreak, even though there were signifi
cant percentages of children at the other day care centers with both diar
rhea and Giardia. This suggests that Giardia is at least a partial cause of the
outbreak at the day care center. The lower percentages of Giardia-free chil
dren with various clinical symptoms suggest a strong relationship between
this pathogen and the diarrheal outbreak. However, other pathogens that
Parasites can Survive inMore than One Host Species 31
70
60 = percent with diarrhea

= percent with Giardia
percentage in children
50
40
30
20
10
0
A day-care day-care surrounding
with outbreak without outbreak community
20
= Giardia (+) family members
= Giardia () family members
percentage in children
15
10
0
B Giardia (+) child Giardia () child
Figure 3 Prevalence of diarrhea and Giardia in a human population.
A, Percentages in children from day care centers and surrounding
community. B, Giardia positive and negative family members with
children in day care center with diarrheal outbreak.
Source: From Black et al., 1977, Table 1 and text.
cause diarrhea seem to be common, based on the prevalence of some clini

cal symptoms in children that were negative for Giardia.
The percentages of children with Giardia were much higher in those
who attended day care than the 2.4% in age-matched children in the
same community who did not attend day care. G. lamblia commonly
infected children even in day care centers that were not suffering diarrheal
60 p < 0.04
50
= percent Giardia (+) children
percentage of children
= percent Giardia () children

40
p < 0.01
30
p < 0.05
20 NS
10 NS NS NS
NS
NS
0
ea
ys
ng
ps
ea
ou th
ve
os
nc
uc i
da
am
m w
s
rh
iti
us
fe
tl
le
m
ar
d/ ea
na
gh
10
tu
cr
vo
di
oo rrh
fla
ei
r>
bl ia
fo
d
ea
rh
ar
di
Figure 4 Percentages of clinical symptoms in children tested for

Giardia. The p-value is the probability of seeing such different
percentages in the two samples, assuming the two conditions are
equivalent. NS, not statistically different.
Source: From Black et al., 1977, Table 2.
outbreaks. The lack of Giardia in children in the community strongly

implicates the day care centers as the source of transmission.
Other data that the scientists collected in the parent interview in
cluded the clinical symptoms manifested by both Giardia positive and
negative children (Figure 4), the prevalence of pet ownership, and any
recent travel. There were no differences in the frequency of pet owner
ship or recent interstate or foreign travel. Finally, the scientists found no
evidence of Giardia contamination in the tap water at the day care center
that had the diarrheal outbreak.
Because there were no differences in pet ownership or recent travel,
the epidemiologists, who study factors affecting health and illness in
populations, were able to eliminate animal-to-human transmission and
infections caught while travelling. Tap water was not contaminated at
the day care center with the outbreak, eliminating drinking water as the
source of the Giardia. The researchers concluded that transmission of
Giardia occurred through direct person-to-person contact and possibly

from contaminated objects at the day care center. They further concluded
that Giardia occurred in a high percentage of children, because they were
either not toilet-trained or were just learning, and they do not always have
good hygiene practices. The fecal-to-oral transmission route was deter
mined to be a viable route for Giardia infections, and children with dirty
hands in close contact can easily pass this pathogen between each other.
The preceding research shows that Giardia cysts can be found in feces.
In another study of day care centers, Larry Pickering and his colleagues
examined the prevalence of Giardia among children up to 2 years old.
The researchers used the same techniques to determine the presence of
Giardia in fecal samples as Black and his colleagues did, this time collect
ing just one fecal sample per child, but from a larger number of children
from 29 day care centers and a second survey 6 months later of 26 of the
same centers (Figure 5).
One major difference in the two studies is that Pickering and his col
leagues examined feces not only for cysts but also for the active, motile
feeding stage of Giardia, called trophozoites.
Pickering and his colleagues found that Giardia occurred consistently
over time among children in day care centers. The two surveys separated
by 6 months suggest a steady level of infection. Cysts are more common
30 Giardia prevalence
25
= first survey
percentage of children
= second survey
20
15
10
0
percentage with cysts percentage with trophozoites
Figure 5 Percentage of children in day care centers with Giardia
cysts and trophozoites.
Source: From Pickering et al., 1984, Table 1.
in feces, suggesting they are produced by the population of Giardia in the

host and released into the environment, to infect other hosts. The tro
phozoites are not common in fecal matter, suggesting that they live inside
the host and are not typically passed out in the feces. Once infected, the
host begins to produce feces infected with Giardia cysts, and exposure to
infected fecal matter may be one way to contract Giardia.
J. Carl Craft performed an experiment to determine the infectivity of
human fecal matter to rats. Over the course of a year, he collected feces of
humans who were sick with Giardia. Once collected, he added distilled
water to the samples, and then stored them in a refrigerator for up to
1year. He pooled the samples together and suspended the cysts in a salt
solution.
Craft fed cysts to ten groups of 3 to 4 week old rats and evaluated them
on day 7 for infection. Each group of ten rats was fed a different number of
cysts, ranging from 50 to 500. Craft showed that G. lamblia, once thought
to be specific to humans, can infect rats, and as few as 50cysts could cause
infection. All rats in every treatment with 150500 cysts became infected,
while only 50% were infected with 50 cysts.
The scientist also fed 50 5-day-old rats a suspension containing about
250 cysts and collected feces daily for 70 days. Feces were examined for
Giardia cysts. Infection occurred in as little as 2 days and persisted for
several weeks in some rats. By day 4 all rats were infected. After 40 days
all rats had cleared the infection.
Craft also killed infected rats with lethal injections of sodium pento
barbital and dissected out 2 cm segments of the duodenum, jejunum, and
ileumthe beginning, middle, and final portions of the small intestine
starting at the lower end of the stomach, respectively. He examined the
contents of the intestinal lumen under a microscope for evidence of cysts
and trophozoites of Giardia. In this experiment, Craft did not quantify
the proportion of trophozoites and cysts in the intestinal sections, but
he did identify trophozoites in the intestines of some rats that were not
passing cysts in the feces. Cysts that infected rats gave rise to trophozoites,
supporting the notion that the trophozoites and cysts are two stages of the
Giardia life cycle.
The scientist also tested the infectivity of cysts of different ages and
determined that cysts that had been stored for a year could still cause
infection in rats, although the number of cysts needed to cause an in

fection was higher than for freshly collected cysts. The observation of
continued infectivity of cysts stored in liquefied feces for 1 year is another
important aspect of their life cycle. If cysts are passed in the feces and then
must be ingested by another host, they must be able to survive outside of
hosts in a potentially harsh environment.
To accurately assess the distribution of trophozoites and cysts in in
testines, J. D. Campbell and G. M. Faubert performed an experiment
onmale gerbils (Meriones unguiculatus). The scientists grew cultures of
G.lamblia trophozoites, which they used to infect forty gerbils. Every
3 days they killed four gerbils and dissected out their small and large
intestines. As with Crafts study, the researchers divided the small intes
tine into three sections. Each section was slit lengthwise, Giardia cells
were detached from the inner surface of the intestines, and samples were
microscopically examined for trophozoites, encysting trophozoites (tro
phozoites with prominent cytoplasmic vesicles), and cysts. The scientists
counted cells in three small samples for each intestinal section, and then
scaled up their counts to determine the abundance of cells in the entire
section.
The experiment by Campbell and Faubert adds one more species to
the growing list of species that G. lamblia can infect. The scientists gave
gerbils a large dose of Giardia trophozoites to quickly infect them and
observed development of the population within the host. The host is the
ecological system in which the parasite population finds itself. The tro
phozoites of Giardia inhabit intestines, and after 6 days are beginning to
develop into encysting trophozoites and then develop into cysts, which
are more abundant in the jejunum, ileum, and large intestine. Only
sporadically do cysts appear in the duodenum. So as the population of
parasites grows in the host, encysting trophozoites move through the in
testine, divide, and change into cysts. Although trophozoites are found in
the large intestines, the majority of Giardia individuals in that section are
cysts or developing into cysts.
Gerbils are susceptible to Giardia, and as in rats and humans, the
infection is cleared out in a matter of a few weeks, due to a response from
the host immune system. Continuing tension between host and parasite
causes evolution of mechanisms in the parasite that facilitate transmission
to new hosts. The development of cysts is a mechanism for transmission,

and cysts move through the intestines and are found in fecal matter. Once
ejected in feces, the cysts must find their way to a new host. Fecal-to-oral
transmission can occur through unhygienic practices, such as may occur
in day care centers. Are there other routes that could lead to transmission
between hosts that are different species?
Alan Barbour and his colleagues from the CDC studied a group of
52students and two teachers who hiked and camped in a remote moun
tainous area in Utah. The goal of the campers was to live off the land as
much as possible for 2 weeks.
Six students reported diarrhea at the university health center upon
their return to campus. There was a spike in the number of new cases
per day by day 12 and this continued for several more days, even after
the campers returned from the trip. They were diagnosed with Giardia in
their feces, and in response the researchers interviewed 53 of the 54 trip
participants.
The time course of onset of new cases is not unlike the occurrence of
Giardia in other animals in the earlier studies. The campers were ques
tioned about onset of any symptoms, including diarrhea, any prior travel,
and their food and water consumption. Giardia can be in the form of
either a trophozoite or a cyst and cysts are more commonly found in the
large intestine and feces than trophozoites. Cysts appear to be an adapta
tion for dispersal of Giardia from a host to the environment and then on
to a new host. Once a cyst is ingested, environmental cues inside the host
trigger developmental changes, which cause cysts to become trophozoites.
Trophozoites living in the small intestine cause illness; cysts can survive
outside the host and are the route of transmission to a new host. The
campers that left for the wilderness trip were not sick prior to leaving,
although they were not known to be definitively negative for Giardia.
Forty-two of the campers submitted fecal samples for Giardia and
bacterial pathogen testing. The researchers defined a positive case as:
1)an illness that lasted 5 days or more accompanied by diarrhea or the
combination of nausea, cramps, bloating or flatulence, and weight loss;
or 2)a fecal sample positive for Giardia. No campers tested positive for
other bacterial pathogens, but 79% tested positive for Giardia. Many had
diarrhea (85.3%), bloating, belching, or flatulence (97.1), nausea (79.4),
and weight loss (55.9). Many symptoms observed in children in day care
centers experiencing Giardia outbreaks appeared in the campers. The
onset of new cases (especially the spike at the end of the camping trip)
strongly suggests that the outbreak was caused by a parasite that infected
the campers while on the trip.
The researchers retraced the steps of the campers and searched for
Giardia cysts in stream water from which the campers drank. All campers
drank from the same water source, but they broke up into four groups to
search for food, with each group consuming what they had collected. The
scientists filtered stream water to concentrate any cysts on the filters. They
found no Giardia cysts. The only common source that all campers were
exposed to was the water, because groups of campers ate food that was not
shared among them, and campers from all four food-collecting groups
got sick. However, the researchers did not find cysts in the stream or in
beavers or muskrats living in streams. Thus there is no direct evidence
of Giardia in the water or in wild animals using the stream as habitat.
However, cysts are difficult to find in contaminated water, because they
are strongly diluted in such sources. Cysts can live outside of a host for up
to a year as shown previously, and the stream water was the only source
of water.
The scientists also examined the small and large intestines of ten
beavers and seven muskrats trapped in the drainage area, and found no
Giardia in the animals. Although evidence of Giardia in wild animals living
in the area might have been expected, the sampled beavers and muskrats
were a small proportion of these populations, and these individuals might
happen to not be infected or the species are not suitable hosts for Giardia.
Brett Dunlap and Monte Thies trapped and killed beaver (Castor ca-
nadensis) and nutria (Myocastor coypus), two semi-aquatic rodents, to pre
vent them from causing damage to human property. The beaver is native
to North America, whereas the nutria is an exotic species indigenous to
South America. The scientists collected a small intestinal and a fecal sam
ple from the large intestine from 100 beavers and 30 nutria. The samples
were preserved and later examined for the presence of Giardia using stain
ing and microscopy techniques similar to those used earlier (Figure 6).
Giardia is common in many mammal species, so it is not surprising
to find it in beavers and nutria. Dunlap and Thies did not distinguish
100
80
sampled with Giardia
percent of animals
60
40
20
0
beaver nutria
Figure 6 Prevalence of Giardia in beaver and nutria captured
ineastTexas.
Source: Data from Dunlap and Thies, 2002, Tables 1 and 2.
different species of Giardia in the samples that they examined, so they

could not conclude that it was G. lamblia that was infecting these ani
mals. However, other studies have shown that beaver are susceptible hosts
for G. lamblia, the species that infects humans. Beavers defecate in or near
the water, and this provides a mode of transmission for Giardia cysts. An
animal that ingests cysts as it drinks may become infected. This is a com
mon way for campers to acquire Giardia.
The pathogen Giardia infects multiple hosts, and it has evolved a com
plex life cycle with different stages to survive in particular environments
either in or out of the host. This unicellular eukaryotic parasite causes
disease in multiple species of animals, and it is an important component
of the ecological system in which it lives. More than a few unicellular
parasites that cause disease in humans can also live within other host
species. Giardia has thus evolved to disperse from host to environment to
host. When a new parasite or pathogen enters a host population, disease
can quickly spread, as seen in the group of campers. More about how
diseases spread will be explored in the next chapter.
Bibliography
Barbour AG, Nichols CR, Fukushima T: An outbreak of giardiasis in a
group of campers, Am J Trop Med Hyg 25(3):384389, 1976.
Black RE, Dykes AC, Sinclair SP, et al.: Giardiasis in day-care centers:
evidence of person-to-person transmission, Pediatrics 60(4):486491,
1977.
Campbell JD, Faubert GM: Comparative studies on Giardia lamblia
encystation in vitro and in vivo, J Parasitol 80(1):3644, 1994.
Craft JC: Experimental infection with Giardia lamblia in rats, J Infect Dis
145(4):495498, 1982.
Dunlap BG, Thies ML: Giardia in beaver (Castor canadensis) and nutria
(Myocastor coypus) from east Texas, J Parasitol 88(6):12541258, 2002.
Pickering LK, Woodward WE, DuPont HL, et al.: Occurrence of Giardia
lamblia in children in day care centers, J Pediatr 104(4):522526, 1984.
CHAPTER 4
Diseases Spread Through

Populations of Susceptible
Hosts
There are many species interactions in ecological systems. In parasite-

host interactions, populations of animals or plants may be susceptible
to newly-introduced disease-causing organisms. The pathogen, the agent
that causes disease, may be a unicellular organism, such as Giardia or
a bacterium. How some parasites cause disease (such as, Giardia and
B.burgdorferi, which causes Lyme disease) and how that affects individu
als and populations has been explored in previous chapters. In this chap
ter, how a newly introduced pathogen spreads among the individuals in a
population will be examined.
Occasionally, new diseases emerge in a population. In late 2002, a
farmer in Guangdong Province, in southern China, was treated for pneu
monia with flu-like symptoms. The patient died, but no definitive diag
nosis was made on the cause of death. By early 2003, many more cases
were observed. The illness usually began with high fever and mild re
spiratory symptoms, but progressed to pneumonia in a few days. The
disease was named severe acute respiratory syndrome (SARS). Because of
the outbreak and severity of the disease, scientists quickly went to work
to discover the causative agent of SARS, which turned out to be a coro
navirus (Figure 7).
Coronaviruses are enveloped by spherical spike proteins with surface
projections that form a corona around the virus and a ssRNA containing
(in the case of the SARS coronavirus) 13 genes that code for 14 proteins.
Y. Guan and his colleagues studied the epidemiology of the first out
breaks in Guangdong Province by gathering data from clinical records of
envelope
ssRNA + nucleocapsid
matrix
spike
Figure 7 The SARS coronavirus, showing the single-stranded RNA

(ssRNA) and four structural proteins: spike, which forms the corona;
envelope; matrix; and nucleocapsid.
Source: http://commons.wikimedia.org/wiki/File:Coronaviruses_004_lores.jpg. Content
Providers(s): CDC/Dr. Fred Murphy. This media comes from the Centers for Disease Control
and Preventions Public Health Image Library (PHIL), with identification number #4814. Public
domain.
patients admitted to hospitals with a diagnosis of atypical pneumonia,

meaning that it was not caused by the bacteria Streptococcus pneumonia. In
addition, the researchers used local health authority reports and doctors
interviews to assess the spread of the disease.
The researchers defined a case as a patient running a high fever
(>38C) with coughing or difficulty breathing who was either in close
contact with a person suspected of having SARS or lived or traveled in an
area known to have recent local transmission of SARS. Any patients that
died from an unexplained acute respiratory illness who had come in close
contact or traveled in an area known to have SARS cases were defined as
a suspected case.
The disease was found in 6 cities within the first 3 months and a
major outbreak of over 200 cases was recorded in Guangzhou in early
February. Out of 305 cases during this early outbreak, 105 of the patients
were healthcare workers. Patients suffering complications were sometimes
Diseases Spread Through Populations of Susceptible Hosts 43
brought from rural outlying cities to Guangzhou for better medical care.
Many of these initial patients were known to be related to one another.
One SARS patient, admitted on December 17, 2002, was a chef who
worked at a restaurant in Shenzhen. This chef came into regular contact
with live caged animals used as exotic game food. He was the second
confirmed case of SARS.
The scientists also studied 55 patients admitted to a hospital in
Guangzhou, Guangdong Province, in early 2003 with atypical pneumonia.
These patients ran fevers for an average of 11.4 days ( 6.8 days, standard
deviation) and developed pneumonia within 4 days of being admitted.
Forty-one of 55 were known to have had definitive contact with another
SARS patient, and 27 were healthcare workers. Guan and his colleagues
examined mucus and blood serum samples for the SARS coronavirus
antibodies. For 22 of the patients, the scientists tested paired samples,
the first either 3 to 5 or 7 to 10 days after disease onset, and the second
15days after onset. Twenty (91%) of these patients tested positive for
SARS antibodies, and 28 (85%) of the remaining 33 patients tested only
once also were found positive for SARS antibodies. Four of the pneumo
nia patients who did not test positive for the SARS coronavirus antibod
ies were later discovered to have influenza. The researchers also tested
60healthy adults; none tested positive for SARS coronavirus antibodies.
The study of the initial outbreak is an important part of epidemiology
and can pinpoint the origin of an epidemic, identify modes of transmis
sion, and identify possible origins of the pathogen. In the southern China
SARS outbreak of 2002, several observations were made by epidemiolo
gists that were clues to origins and modes of transmission. One observa
tion was that the second known SARS patient was a chef who came into
regular contact with live caged animals. The epidemiological investigation
discovered that his wife, two sisters, and seven healthcare workers all be
came infected from him.
In fact, 34% of all patients in the outbreaks in the initial six cities were
healthcare workers. A patient in Guangzhou who spent only 18 hours in a
hospital in Zhongshan infected 30 healthcare workers in Zhongshan and
Guangzhou. This patient also infected 19 family members or relatives in
Guangzhou. This large number of infections in relatives and healthcare
workers gave rise to the Guangzhou outbreak. One physician infected
during this time traveled to Hong Kong and became the source of the
SARS outbreak that occurred later in Hong Kong. Because the outbreaks
were characterized by infections in hospitals and family units, Guan and
his colleagues concluded that transmission occurs through close contact
with infected patients. The spread from city-to-city occurred because of
transport of patients to better hospital facilities and movement of workers
from city-to-city. This is comparable to a parasite like Giardia, which is
transmitted through fecal-oral contact or contaminated water.
Close person-to-person contact appears to be the mode of transmis
sion for SARS, where coughing or sneezing spreads small droplets up to
about a meter from infected persons, and disperses the virus. The virus
only needs to land on the mouth, nose, or eyes of a nearby person for
them to become infected. If droplets land on another persons hands, or
if an uninfected person touches a surface contaminated with droplets
and then touches their mouth, nose, or eyes, they can become infected.
Close contact may also mean kissing or hugging between an infected
and uninfected person. These are thought to be the most likely routes of
transmission.
Most patients with atypical pneumonia that were tested for the SARS
coronavirus had it as shown earlier. Several were infected with a strain of
the influenza virus. Given the large percentage of patients in this study
with the SARS coronavirus, Guan and his colleagues concluded that the
outbreaks were caused by this newly emerging viral pathogen. The scien
tists were able to conclude that the pathogen was emerging because none
of the healthy patients had antibodies to the SARS coronavirus, which
suggests that the virus was not previously present in this human popula
tion. The researchers, in attempting to track down the origin of the SARS
coronavirus, focused on the second known SARS patient, the chef, and
the observation that he came into contact with exotic game animals. In
China and many other countries, wild animals are caught or raised in cap
tivity to be sold live at markets for human consumption. Some wild game
animals are considered a delicacy in southern China and are bought by
chefs for preparation at their restaurants. Other diseases are also known
to spread from animals to humans.
Guan and his colleagues set out to test the hypothesis that the SARS
coronavirus was present in wild animals sold at southern Chinese markets.
The researchers tested a variety of both domestic and wild mammals that
might be carrying the SARS coronavirus in Guangdong Province. The 25
animals sampled were obtained from a live animal retail market in Shen
zhen and included seven wild and one domestic mammal species.
The animals came from different areas of Guangdong and had been
kept in separate storehouses prior to coming to market. Only a few ani
mals of each species were kept by any one stall owner. Guan and his
colleagues obtained nasal, fecal, and blood (when possible) samples from
animals housed in different stalls. The sampled animals were confirmed
to have no overt symptoms of disease by a veterinarian. The researchers
used reverse transcription polymerase chain reaction (RT-PCR) to test
for the SARS coronavirus. In RT-PCR RNA is reverse transcribed into
its DNA complement and detected by binding it to a primer specific to
the human SARS coronavirus. In addition, the scientists also tested for
presence of the virus by inoculating the samples into cell cultures. If the
virus was present, the cells displayed degenerative changes associated with
the multiplication of viruses.
The beaver (Castor fiber), Chinese ferret-badger (Melogale moschata),
Chinese hare (Lepus sinensis), deer (Chinese muntjac, Muntiacus reevesi),
domestic cat (Felis catus) and hog-badger (Arctonyx collaris) never tested
positive in 18 total samples. However, all of the Himalayan palm civet
(Paguma larvata), raccoon dog (Nyctereutes procyonoides), 7 out of 7 ani
mals, tested positive. The Himalayan palm civet is a small carnivore found
throughout Southeast Asia and the raccoon-dog is a type of dog found in
eastern Asia.
Guan and his colleagues also collected blood samples from 20 wild
animal traders, 15 butchers, and 20 vegetable retailers. They tested these
samples for antibodies to the virus isolated from one of the palm civ
ets (Paguma larvata) using an immunofluorescence assay. Prior to being
tested, none of these individuals reported SARS-like symptoms. Higher
percentages of wild animal traders and butchers, individuals handling an
imal products, tested positive for the SARS coronavirus antibody (40%
and 20%, respectively) than vegetable retailers (5%).
Although Guan and his colleagues could not conclude that either the
civet or the raccoon-dog is the reservoir for the SARS coronavirus, its
presence in these market animals suggests a connection between animals
and humans. An epidemiological reservoir is a species that possesses a

pathogen and can pass it to other species. The positive animals could have
been infected from another untested source in the market, so the true
reservoir cannot be unequivocally identified. The scientists did conclude,
however, that culinary practices in southern China may facilitate corona
virus transmission from animal to human. Although the data suggest a
link between animals and humans, it cannot be concluded that the coro
navirus in animals is the one that infected humans. It could have passed
from the humans to the animals, although this is less likely given that a
higher percentage of market workers who deal with animals possessed
SARS coronavirus antibody compared to those working exclusively with
vegetables. More species and more markets would have to be tested and
monitored to determine the true source of the outbreaks.
To further determine the relationship between the coronavirus found
in civets and that found in humans, Guan and his colleagues sequenced
partial or entire coronavirus genomes isolated from four of the civets.
They found that the sequences from civets had 99.8% similarity to those
from humans. The researchers performed an evolutionary tree analysis
with sequences of the spike gene from 15 samples: three from civets,
one from a raccoon-dog, and 11 isolated from humans in Hong Kong,
Guangdong, Canada, and Vietnam.
Based on the high degree of overall genetic similarity observed by
Guan and his colleagues, they concluded that the SARS coronavirus is
found in animals. The viruses isolated from animals in one market were
all more related to one another than any one of them was related to vi
ruses isolated from humans. The viruses isolated from the raccoon-dog
and from one of the palm civets were almost genetically identical. One
important point is that all the human coronavirus samples appeared to
have branched off from one of the samples from a civet. The scientists
could not determine whether the virus was transmitted from one animal
to the other, but given the high percentage in civets, it is likely that the
civet is a reservoir host for the coronavirus.
The researchers noted 38 nucleotide changes, 26 of which caused
changes in the transcribed amino acid sequence of the spike protein. Of
the four animal samples, only eight of the nucleotides were polymorphic
(that is, varied among the species), and only two of them led to altered
amino acid sequence of the resulting protein. Among the humans, twenty
or more of the spike gene nucleotides varied among the patients with
many altering the spike protein. This suggests that the jump from civet
to humans occurred further in the past, because the coronavirus DNA
sequences in humans contained many alterations not observed in the
coronavirus isolated from civets and raccoon-dogs.
Animal viruses isolated from one market should be more similar to
each other than the human viruses isolated from Hong Kong, Guangdong,
Canada, and Vietnam, but this was not the case. The viruses isolated from
humans from five geographically separate sites were very closely related
genetically. Even though the total number of mutations was greater, many
of the same mutations occurred in all human virus samples. This close
relationship among the coronavirus isolates from humans indicates that
the virus had spread very rapidly from China to as far away as Toronto,
Canada, after it mutated and jumped from civets, but faster than it
evolved major new differences. The scientists also concluded that the
Chinese live animal markets may have played a role in amplifying the
SARS coronavirus and transmitting this disease to humans, as suggested
by the evidence reviewed earlier.
Recall that a physician infected by a patient in Guangdong later trav
eled to Hong Kong. With a pathogen like the SARS coronavirus, which
has such effective dispersal abilities, SARS could quickly spread beyond
Guangdong province, which is exactly what it did. The World Health
Organization (WHO) keeps statistics on many diseases and epidemics
and compiled data on cases of SARS from China and beyond (Figure 8).
In addition to the cases shown, over 2,500 probable cases of SARS from
Beijing, China, are not shown because date of onset was not available to
WHO. The date of onset for the initial case in countries that had more
than ten cases of SARS is shown in the figure, along with the number
of cases that ultimately occurred in those countries. Several countries
had outbreak onset on the same date, and the number of cases shown is
pooled for those countries.
As global pandemics go, SARS was short-lived but widespread. A
pandemic is an epidemic occurring over a wide geographic area and af
fecting a high proportion of the population. Within just a few months of
its appearance in Guangdong Province, SARS spread to more than two
1 = China, 5,328
2 = Hong Kong, 1,755
160 3 = Canada, United States,
Vietnam, 341
140 4 = Philippines, Taiwan,
120 Singapore, 598
numner of cases
100 Guangdong
Provice outbreaks
80 3
60 2 4
40 1
20
0
2
03
03
0
l
r
ov
ov
ec
ar
ar
ay
ay
Ju
Ap
Ja
Ju
Ja
Fe
M
N
11
18
3
20
24
20
14
1
22
13
30
date of onset
Figure 8 SARS cases by week of onset. Total number of cases shown
is 5,910 out of 8,096 total cases. Not all of Chinas 5,328 cases are
shown in the graph (see text). The time frame for the Guangdong
Province outbreaks described in text is shown.
Source: Data from World Health Organization. http://www.who.int/csr/sars/ country/
table2004_04_21/en/ index.html.
dozen countries around the world. According to the WHO, a total of

8,098 people worldwide became sick with SARS during the 2003 out
break. Of these, 774 died, which is a 9.6% fatality rate. The rapid increase
in the number of cases was due to the way that SARS spreads through
close contact. Twenty-one percent of all cases were healthcare work
ers, and until epidemiologists knew what they were dealing with, many
healthcare workers did not take proper precautions to prevent infection.
The research performed by laboratories in several different countries
provided information necessary to understand and combat the emerg
ing SARS epidemic. The SARS global pandemic was finally contained
in the summer of 2003 through a combination of information sharing
among public health authorities, adequate health screening of travelers,
quarantine of patients and individuals who came in contact with patients,
and education of healthcare workers and the general public. Rapid action
by national and international health authorities helped slow transmission
and eventually broke the chain of transmission. However, SARS has not
been eradicated, and it could reemerge.
The SARS outbreak illustrates how diseases spread. In the case of
SARS, the emergence of a disease transmitted quickly from one host to
another through close contact led to an epidemic in southern China and
then to a pandemic. A global response was necessary to curb the spread
of the disease. Epidemiologists worked quickly and collaboratively to
study this new disease, which was caused by a pathogen that did not pre
viously infect humans. Pathogens are part of ecological systems and they
can infect multiple hosts. Pathogens can evolve to switch to new hosts,
and this often leads to disease outbreaks. As ecological systems change,
opportunities for new interactions among species arise.
Bibliography
Dandekar AA, Perlman S: Immunopathogenesis of coronavirus infec
tions: implications for SARS, Nat Rev Immunol 5(12):917927, 2005.
Guan Y, Zheng BJ, He YQ, et al.: Isolation and characterization of viruses
related to the SARS coronavirus from animals in Southern China,
Science 302(5643):276278, 2003.
World Health Organization: Global alert and response (GAR): severe
acute respiratory syndrome (SARS), World Health Organization (web
site): http://www.who.int/csr/sars/en/. Accessed July 3, 2014.
Zhong NS, Zheng BJ, Li YM, et al.: Epidemiology and cause of severe
acute respiratory syndrome (SARS) in Guangdong, Peoples Republic
of China, in February, 2003, Lancet 362(9393):13531358, 2003.
Conclusion
The themes of internal environments, communication, structure and
function, and cells coming from preexisting cells have all been illus
trated in this chapter. The focus has been on diseases and their effects
at the cellular and organismal levels. Cell function can be disrupted by
altered DNA, proteins, and by pathogenic organisms. The disruption
affects internal environments or ability to maintain a constant internal
environment. Similarly, cell-to-cell communication may be disrupted or
manipulated by pathogens. A single population of a pathogenic organism
can interact with multiple host species, affecting their ability to interact
with other species. Pathogens can have complex life cycles and cause dis
eases to spread. The study of cells can provide insights into how diseases
affect multicellular organisms. The cell links other levels of the biological
hierarchyeffects at the molecular level are manifested at the cellular
level, which can then affect the entire organism.
Glossary
antibodies. Any of a large number of proteins produced by specialized B cells
after stimulation by an antigen and acting against the antigen in an immune
response, and that typically consist of four subunits.
blast. Term used to describe a syndrome of plant diseases where afflicted leaves
suddenly die.
cells. The smallest structural and functional unit of an organism.
chromatography. A process in which chemicals carried by a liquid or gas are
separated into components as a result of differential distribution of solutes as they
flow over a stationary liquid or solid substrate.
cytokines. Cytokines are small proteins, peptides, or glycoproteins released by
cells involved in cell-cell interaction and communication.
denatured. To modify the molecular structure of a protein or DNA by heat, acid,
alkali, or ultraviolet radiation, and thus destroy or diminish biological activity.
disease. A disease is a condition of an organism that impairs normal functioning,
manifested by distinguishing signs and symptoms.
dispersal. Dispersal means to spread from the place of birth.
ectothermic. Of or relating to an animal that cannot regulate its own body tem
perature, so its body temperature fluctuates according to its surroundings.
electrophoresis. The process in which large molecules can be separated according
to size and electrical charge by applying an electric current to them in a gel.
emerging disease. Emerging diseases are infectious diseases that are new in a
population or are rapidly increasing in incidence or geographic range.
endothermic. Endotherms are animals that use metabolic processes to maintain
a relatively constant body temperature.
eukaryote. An organism whose cell or cells contains a distinct, membrane-bound
nucleus.
exon. Portions of eukaryotic mRNA that are retained during mRNA splicing.
familial ALS. An inherited form of amyotrophic lateral sclerosis (ALS), an incur
able and common form of motor neuron disease, a progressive neurodegenerative
disease that affects nerve cells in the brain and spinal cord.
hemoglobin. Hemoglobin is a protein within red blood cells that carries oxygen
and releases it to cells lacking oxygen.
immunofluorescence. The labeling of antibodies or antigens with fluorescent
dyes for the purpose of demonstrating their presence.
leucocytes. Leucocytes are white blood cells that help the body fight infections
and other diseases.
54 GLOSSARY
life cycles. The stages through which an organism passes from the beginning of
its life until its death.
motor neuron disease. A progressive and often fatal neurodegenerative disease.
motor neurons. Motor neurons convey electrical impulses from the nervous sys
tem to a muscle or gland.
mutations. Mutations are changes in a DNA sequence that may or may not alter
the phenotype.
normal. Conforming to a type or a standard, for instance, of a behavior.
osmotic pressure. Osmotic pressure is the pressure exerted by water through
a semi-permeable membrane separating two solutions with different solute
concentrations.
outbreak. An outbreak is an occurrence of disease greater than would otherwise
be expected in a particular time and place.
pandemics. A global outbreak, or epidemic.
pathogenic. Disease causing.
phagocytic. Describing the function of white blood cells that ingest microbes,
other cells, and foreign particles.
phylum. Phylum is the term used to denote the primary division of a kingdom,
ranked above class in size.
plasmolysis. Plasmolysis is the bursting of a cell due to influx of a large abun
dance of molecules.
pneumonia. A disease that affects the lungs and makes it difficult to breathe.
point mutation. A point mutation is a change in a single nucleotide in a DNA
sequence.
population. A population is a group of individuals of the same species living in
the same place at the same time
proteins. Macromolecules with have shapes that determine their function and are
formed by the assembly amino acids.
reservoir. An epidemiological reservoir is a species that possesses a pathogen and
can pass it to other species.
RNA. Chemically similar to DNA except the ribose retains the 2 oxygen.
ssRNA. single-stranded RNA
tissues. Collections of more than one cell type of a single species that work to
gether to perform a complex function.
trypsin. An enzyme that breaks down proteins.
vector-borne disease. Vector-borne diseases are diseases in which the pathogen is
transmitted to an individual by some other agent.
Index
Allison, Anthony, 78 Fused in sarcoma/translated in
Amyotrophic lateral sclerosis (ALS), 9 liposarcoma (FUS/TLS) gene,
mutations that co-inherited 910
with, 10 mutations affect function of, 11
Anemia, 7 reason for MND, 12
Animals in research, 2224
Arginine, 5 Genetic diseases
ethical, legal, social implications,
Bacteria both, 15 1214
Bacterial plasmid, 19 sickle-cell disease
Barbour, Alan, 36 by multiple mutations, 912
Black, Robert, 29 by single point mutation, 18
Blast, 2425 Gerbils, male, experiment on, 35
Borrelia burgdorferi, 1517 Giardia lamblia, 29, 30, 41, 44
culture of genetically engineered, 19 campers to live off land, 3638
hosts immune system, 22 in children, 3033
cysts in feces, 33
Campbell, J. D., 35 experiment on male gerbils, 35
Castor canadensis, 37 Glutamic acid, 5
Cells burst (plasmolyze), 26 Grimm, Dorothee, 16
Centers for Disease Control and Guan, Y., 41
Prevention (CDC), 29
Chemokines, 18 Hemoglobin, 2
Coronaviruses, 41 in carbon monoxide (CO), 2
animal, 4546 fingerprints, 45
human, 46 Himalayan palm civet, 45
severe acute respiratory syndrome. Host cells, 2728
See Severe acute respiratory Howard, Richard, 25
syndrome (SARS) Human coronavirus, 46
Craft, J. Carl, 34 Human fecal, infectivity of, 3435
delGG, 10 Immunofluorescence assay, 45

Infection cells, pressure built in, 27
Electrophoresis, 2 Ingram, Vernon, 4
protein concentration scanning insGG, 10
in, 3
Epidemiological reservoir, 46 KC plasmid, 19
Exon 5, 10 high doses of, 2122
Familial ALS, 9 Leucocytes, 18

Faubert, G. M., 35 Lou Gehrig disease. See Amyotrophic
Fungi, to be plant pathogens, 2428 lateral sclerosis
56 INDEX
Lyme disease, by vector-borne PEG. See Polyethylene glycol

bacterium, 1522 Perfection, definition of, 1314
Lysine, 5 Phagocytic leucocytes, 18
Phylum, 24
Magnaporthe grisea. See Rice blast Pickering, Larry, 33
fungus Plasmolysis, 25
Malaria parasites, in blood of normal Pneumonia, 43
and sickle-cell children, 8 Point mutations, 6
Male gerbils, experiment on, 35 Polyethylene glycol (PEG) polymers, 25
Mammals cells burst (plasmolyze) exposed to, 26
immune system of, 18 create different levels of internal
infection of, 1819 pressure, 2627
Melanin, 25 pressure that inhibited penetration
cells without, 26 of substrates, 27
fungus on artificial substrates, 25
loss of, 26 Raccoon dog, 45
Meriones unguiculatus, 35 Reverse transcription polymerase
MND. See Motor neuron disease chain reaction (RT-PCR), 45
Monocytes, 18 Rice blast fungus, 2425
Motor neuron disease (MND), 9
Mutations Severe acute respiratory syndrome
by multiple, 912 (SARS), 4143
by single point, 18 epidemiology, 43
Myocastor coypus, 37 global pandemic, 4748
mode of transmission for, 44
Neutrophils, 18 patients with atypical pneumonia
of B. burgdorferi infection, 21 tested for, 44
lack of, 19 reservoir for, 4546
Sickle-cell anemia, 12
Osmotic pressure, 26 Sickle-cell disease
OSP-C negative genome, 16 by multiple mutations, 912
mice infected with, 1718 by single point mutation, 18
ssRNA, 41
Parasites, to cause disease, 2938 Streptococcus pneumonia, 42
campers to live off land, 3638 Survival, growth, and dispersal,
development of cysts, 36 concepts of, 29
experiment on male gerbils, 35
Giardia lamblia in children, 3033 TDP-43 protein, 10
-host interactions, 41 reason for MND, 12
human fecal, infectivity of, 3435 Tick saliva, 15
life cycles of, 29 Trophozoites, 33
staining method, 30 in small intestine, 36
Pathogens, affect cells and organisms, Trypsin, 4
15, 41
animals in research, 2224 Ubiquitin, 1112
fungal, 2428 Unicellular eukaryote, 29
Lyme disease, by vector-borne
bacterium, 1522 Vector-borne bacterium, Lyme disease
Pauling, Linus, 2 by, 1522
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EBOOKS Effects of Genetic and Pathogenic

FOR THE Diseases onCells

Christopher J. Paradise, PhD

All rights reserved. No part of this publication may be reproduced,

First published in 2016 by

ISBN-13: 978-1-60650-961-6 (print)

Momentum Press Biology Collection

Cover and interior design by S4Carlisle Publishing Services Private Ltd.,

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Genetic Diseases Affect

Genetic diseases are caused by mutations. Mutations are changes in the

Sickle-Cell Disease is Caused by a Single Point

pressures. They typically suffered no long-lasting effects from this phe

Table 1 Movement of normal and sickle-cell hemoglobin at different pHs.

A way of scanning the protein concentration in an electrophoresis

Table 2 Amino acid sequence alignment for the peptide hemoglobin

Table 3 Solubility (g/L) of normal and sickle-cell hemoglobin at

whereas that of deoxygenated normal hemoglobin is 1.6 g/L and that of

Table 4 Incidence of the malaria parasite in blood of normal and

under the age of 5 in a single Ugandan community where malaria was

Multiple Mutations may Cause a Single Disease

modified, and transported in order to make proteins. It happens to link

binds to RNA, and regulates how messenger RNA is created, modified,

proteins being tagged by ubiquitin. Usually the FUS/TLS protein func

Ethical, Legal, Social Implications:

essence, like the chemical elements. That would make each of us im

Pathogens Affect Cells

Lyme Disease Is Caused by a Vector-Borne Bacterium

and arthritis, numbness in the extremities, and cognitive problems. The

To determine whether mutant bacteria could infect mammals, Grimm

performed a study to determine if this lack of neutrophil attraction was a

percent of 200 cells that were neutrophils or monocytes

Table 5 A, Percentages of tissues from ten mice injected with either

By artificially attracting neutrophils to the site of a B. burgdorferi

Ethical, Legal, Social Implications: There are

has and can be gained from performing research on animals. Researchers

Fungi can be Pathogens of Plants

cover leaves. Some individuals are resistant to infection; those plants de

Table 6 Response of rice blast infection cells when exposed to

Cells burst (plasmolyze) when exposed to PEG molecules small enough

Parasites can Survive

Populations of organisms interact with each other, and one common in

Figure 2 Giardia lambla trophozoite.

60 = percent with diarrhea

cause diarrhea seem to be common, based on the prevalence of some clini

= percent Giardia () children

Figure 4 Percentages of clinical symptoms in children tested for

outbreaks. The lack of Giardia in children in the community strongly

Giardia occurred through direct person-to-person contact and possibly

in feces, suggesting they are produced by the population of Giardia in the

infection in rats, although the number of cysts needed to cause an in

to new hosts. The development of cysts is a mechanism for transmission,

different species of Giardia in the samples that they examined, so they

Diseases Spread Through

There are many species interactions in ecological systems. In parasite-

Figure 7 The SARS coronavirus, showing the single-stranded RNA

patients admitted to hospitals with a diagnosis of atypical pneumonia,

and humans. An epidemiological reservoir is a species that possesses a

dozen countries around the world. According to the WHO, a total of

delGG, 10 Immunofluorescence assay, 45

Familial ALS, 9 Leucocytes, 18

Lyme disease, by vector-borne PEG. See Polyethylene glycol

Behavior and Information Exchangeby Christopher J. Paradise and A. Malcolm Campbell