Lecture 1: Intro to Microbiology

❖​ Pseudopods
Chapter 1 ❖​ HIVs
The Microbial World and You

What is Microbiology? History of Microbiology

The branch of biology which dealing with the study ❖​ Ancestors of bacteria were the first life on
of microorganisms, or microbes( a diverse group of Earth.
minute, simple life forms that include bacteria, History of Microbiology
archaea, algae, fungi, protozoa, and viruses). The
field is also concerned with the structure, function, ❖​ 4.55 b.y.a: earth forms
and classification of such organisms and with ways
of both exploiting and controlling their activities. ❖​ 4.2-4.1 bya: crust forms

❖​ 4.0bya: comets: meteors, asteroids continues

Microbes, or microorganisms are very small living
things that are usually can not be seen with the to bombard surface
naked eye. ❖​ 3.85 bya: first traces of biochemical

Most of them are harmless (99%), but some are ❖​ 3.7bya: oldest fossils
pathogenic (1%) microorganisms which can
affect human, animals, plants and other ❖​ The early Greeks believed that living things
microorganisms. could originate from nonliving matter
(abiogenesis) (the goddess Gea could create
“Germ” refers to a rapidly growing cell. life from stones).
❖​ Aristotle discarded this notion, but he still
Advantages of microbes ?
Microbes have many important and beneficial heldthat animals could arise spontaneously
Biological functions fromdissimilar organisms or from soil.
- Decompose organic waste (recycle nutrient) (spontaneous generation)
- Producer in the ecosystem ❖​ This concept was still felt as late as the
(algae, and some bacteria) capture the energy
from sun light and convert it to food, forming the 17thcentury, but toward the end of that century
basis of food chain (photosynthesis) a several observations, experiments, and
arguments began that eventually refuted this
--Food industry: Produce fermented foods such as concept.
vinegar, cheese, yogurt and bread. ❖​ In 1665, Robert Hooke (Englishman) reported
- Other industries: Produce industrial chemicals that living things were composed of little boxes
such as ethyl alcohol and acetone. or cells using a microscope (was the first to use
-Nitrogen fixation: some bacteria can take the a microscope to observe living things).
nitrogen from air and incorporate it into soil.
- Medicine and medical research: many antibiotic ❖​ Antonie van Leeuwenhoek (1632–1723)
and other drugs are naturally synthesized by (Dutch)
microbes (penicillin made by molds).
-Digestion: Humans and animals are have ❖​ Microbiology essentially began with the
microorganisms in their digestive tract that are development of the microscope.
essential for digestion, and overall health ❖​ Although others may have seen microbes
* Therefore, microbes are essential for life in the before him, Antonie van
earth. ❖​ Leeuwenhoek (1632–1723) was one of the first
people to observe microorganisms (1675) , using
The importance of study the microbiology
a microscope of his own design.
-Description of protozoa, basic types of bacteria,
The study of microbiology provide us with:
yeasts, and algae.
-​ To understand how they affect our lives
- observed and described live microorganisms
and how we can exploit them.
in teeth scrapings, rain water. peppercorn
-​ Knowledge of Microbes allows humans to
infusions and faces as Animalcules
prevent microbial disease and food
spoilage.
So now there are two hypotheses:
-​ Led to aseptic techniques to prevent
The hypothesis that living organisms arise from
contamination in medicine, food industry
nonliving matter is called spontaneous
and in the microbiology laboratories
generation. According to spontaneous generation,
a “vital force’ Forms life.
Example of microorganisms
❖​ Bacteria The Alternative hypothesis, that the living
organisms arise from preexisting life, is called
❖​ Sporangia biogenesis.
 was not hot enough to evaporate the alcohol in
❖​ Francesco Redi (1626-1697) (Italian)
wine. This application of a high heat for a short
- Performed an experiment to disprove time (30 mins) is called pasteurization.
spontaneous generation.
- He showed that the maggots would not arise from Contribution of Louis Pasteur
decaying meat, when it is covered • He coined the term “ Microbiology”
- He set up an experiment to disprove • He disproved the Spontaneous generation Theory
spontaneous generation of maggots. Filled 2 jars • He demonstrated that anthrax was caused by
with decaying meat, sealed one and left the other bacteria.
open (only the open jar developed maggots). • He invented the process of pasteurization and
❖​ Another experiment was set up in which a jar fermentation.
• The development of effective vaccine
was covered with a fine mesh instead of being (rabies and anthrax)
sealed so that fresh air could enter the jar • Pasteur works along with subsequent
again, only the open jar developed maggots) observations by Lister, Koch and others established
Proved that the flies had to come in contact the germ theory of disease
with the meat and lay their eggs on it, maggots
did not arise spontaneously. The Germ Theory of Disease
❖​ Redi filled six jars with decaying meat. 1835: Agostino Bassi showed a silkworm
Conditions Results disease was caused by a fungus.
3 jars covered with No maggots 1865: Pasteur believed that another silkworm
fine net disease was caused by a protozoan.
3 open jars Maggots appeared 1840s: Ignaz Semmelwise Advocated washing
From where did the maggots come? hands to stop the spread of disease.
What was the purpose of the sealed jars? 1860s: Joseph Lister used a chemical
Spontaneous generation or biogenesis? disinfectant to prevent surgical wound infections
after looking at Pasteur’s work showing microbes
❖​ 1861: Louis Pasteur (Father of medical are in the air, can spoil food, and cause animal
bacteriology) demonstrated that diseases.
microorganisms are present in the air. 1876: Robert Koch provided proof that a
​ Conditions Results bacterium causes anthrax and provided the
Nutrient broth placed Microbial growth experimental steps, Koch’s postulates, used to
in flask, heated, not prove that a specific microbe causes a specific
sealed disease.
Nutrient broth placed No microbial growth ❖​ Koch was a physician and Pasteur’s young
in flask, heated, then
sealed rival
Spontaneous generation or biogenesis? ❖​ Koch's Postulates
-​ are used to prove the cause an infectious
❖​ Next experiment, Pasteur’s S-shaped flask disease
kept microbes out but let air in. These -​ are a sequence of experimental steps to
experiments form the basis of aseptic relate a specific microbe to a specific
technique disease.

1892 Dmitri Iosifovich Ivanovski : Discovered

❖​ The Golden Age of Microbiology 1857-1914 viruses
A young milkmaid informed the physician Edward
❖​ Beginning with Pasteur’s work, discoveries Jenner that she could not get smallpox because
included the relationship between microbes she had already been sick from cowpox.
and disease, immunity, and antimicrobial 1796: Edward Jenner inoculated a person with
drugs cowpox virus. The person was then protected from
❖​ Pasteur showed that microbes are responsible smallpox.
-​ Called vaccination from vacca for cow
for fermentation. -​ The protection is called immunity
❖​ Fermentation is the conversion of sugar to
What can you say about the cowpox and
alcohol to make beer and wine.
smallpox viruses?
❖​ Microbial growth is also responsible for
spoilage of food. Vaccinations
-​ produced from avirulent microbial strains
❖​ Bacteria that use alcohol and produce acetic -​ produced from live viruses
acid spoil wine by turning it to vinegar (acetic -​ produced from viral particles
acid).
Chemotherapy – treatment with chemicals
Chemotherapeutic agents used to treat infectious
❖​ Pasteur demonstrated that these spoilage
disease can be synthetic drugs or antibiotics.
bacteria could be killed by heat (62.8 c) that
Antibiotics are chemicals produced by bacteria Lecture 3
and fungi that inhibit or kill other microbes. Morphological and Physiological
Quinine from tree bark was long used to treat Characteristic of Microorganisms
malaria.
Morphological
Chemotherapy – treatment with chemicals Microbes can be classified into four major groups:
1910: Paul Ehrlich developed a synthetic arsenic 1- Protozoa 2- Bacteria.
drug, salvarsan, to treat syphilis. 3- Fungi. 4- Viruses.
1930s: Sulfonamides were synthesized.
1928: Alexander Fleming discovered the first 1- The Protozoa: These are unicellular organisms
antibiotic. with protoplasm differentiated into nucleus and
-​ He observed that Penicillium fungus made cytoplasm.
an antibiotic, penicillin, that killed -​ Diameters in the range of 2-100 μm.
Staphylococcus aureus. The most important groups of medical protozoa
1940s: Penicillin was tested clinically and mass are:
produced. A-Amoeba: Entamoeba species. Mode of Motility:
pseudopodia.
B- Mastigophora: Mode of Motility: the Flagella.
❖​ Modern Developments
Gastrointestinal flagellates: Giardia intestinalis
Urogenital flagellates: Trichomonas vaginalis
Bacteriology is the study of bacteria.
Mycology is the study of fungi. Tissue and blood flagellates: Trypanosoma,
Parasitology is the study of protozoa and Leishmania .
parasitic worms. Trypanosoma, and Leishmania .
Recent advances in genomics, the study of
an organism’s genes, have provided new C- Ciliophora: motile by cilia.
tools for classifying microorganisms. Example: Balantidium coli.
D- Sporozoa: intracellular infection.
Taxonomy Example: Plasmodium that cause Malaria.
• The science of classifying organisms
• Provides universal names for organisms 2- The bacteria:
• Provides a reference for identifying Bacteria are unicellular prokaryotic microorganisms
organisms that multiply by binary fission.
-​ Bacteria can be classified according to
Classification of Microbes morphology, arrangement, and staining
Taxonomy reaction into the following groups:
• Systematics or phylogeny 1- Filamentous bacteria: Streptomyces: antibiotic
• The study of the evolutionary history producers.
of organisms 2- True bacteria:
• All Species Inventory (2001-2025) Cocci: Gram positive: Staphylococcus, Streptococcus.
• To identify all species of life on Earth Gram negative: Neisseria.
Taxonomic Hierarchy Bacilli: Gram positive: Bacillus, Clostridum, Corynebacterium.
Domain Binomal Gram negative: Enterobacteriaceae, Brucella.
Nomenclature uses 3- Spirochetes: Slender flexuous spiral bacteria.
Kingdom Borrelia, Treponema, Leptospira.
the Genus and 4- Mycoplasma: The Smallest bacteria that lack of
Phylum Species name to a rigid cell wall.
Class identify each 5- Rickettsiae and Chlamydiae: intracellular
creature. parasites.
Order
Family Structure of bacterial cells:
Genus Size, Shape, and Arrangement of bacterial cells:
Species Morphology and arrangement of bacterial cells are
criteria used for classification of bacteria into
following groups:
Taxonomic Hierarchy 1. Cocci (Singular: coccus).
Each name is Latinized 2. Rods (bacilli), (Singular: rod, bacillus).
There is a specific way to write each name. 3. Vibrios (Singular: vibrio).
Homo sapiens 4. Spirilla (singular :Spirillum)
The first word is capitalized 5. Spirochetes. (Singular: Spirochaete).
Name is in italics
Homo sapiens 1. Cocci:
H. sapiens - These are round or oval bacteria measuring about
0.5-1.0 micrometer in diameter. When they
multiplying, cocci may form pairs, chains, or
irregular groups.
-​ Cocci in pairs are called diplococci, for
example, meningococci and gonococci.
 -​ Cocci in chains are called streptococci, for -​ They are obligate intracellular parasites.
example Streptococcus pyogens. -​ They are only seen by electron microscope.
-​ Cocci in irregular groups are called -​ Depend on the parasitized cell for survival
Staphytococci, for example, and multiplication
Staphylococcus aureus.

2. Rods (bacilli):
- These are stick-like bacteria with rounded,
square, or swollen ends.
- They measure 1-10 micrometer in length by
0.3-1.0 micrometer in width.

It may arranged in:

A- Chains, for example, Streptobacillus species.
B- Branching chains, for example, lactobacilli .
C- Mass together, for example, Mycobacterium
leprae.
D- Remain attached at various angles resembling
Chinese letters, for example, Corynebacterium
diphtheria.
Cell Structure
3-Vibrios:
• Two structural types of cells are recognized: the
-​ These are small slightly curved rods
prokaryote and the eukaryote. Prokaryotic cells
measuring 3-4 micrometer in length by 0.5
have a simpler internal structure than eukaryotic
micrometers in width.
cells, lacking membrane-enclosed organelles.
-​ Most vibrios are motile with a single
flagellum at one end.
-​ They show a rapid darting motility.
For example: vibrio cholerae.

4-Spirochetes:
-​ These are flexible, coiled, motile organism,
6-20 micrometer in length.
-​ They progress by rapid body movements.
Spirochetes are divided into three main groups:
A- Treponemes. B- Borreliae. C- Leptospires.

3- The Fungi:
-​ These are saprophytic or parasitic
organisms possessing relatively rigid cell
walls.

Medical fungi can be divided into:

1- Mould: Branching filaments; hyphae, mycelium.
Usually 2 to 10 μm in width.
Example: Epidermophyton, Trichophyton,
Microsporum, Aspergillus.
2- True Yeasts: these are ovoid or spherical cells Bacterial Morphology
that reproduce asexually by budding and sexually
with formation of spores. • Some typical bacterial morphologies include
Example : Cryptococcus spp. coccus, rod, spirillum, spirochete, appendaged, and
3- Dimorphic fungi: Produce a vegetative filamentous.
mycelium in artificial media, but are yeast like in
infected lesions.
Example: Histoplasma.
4- Yeast- like fungi: Example: Candida (
Pseudomycelium).

4- The viruses:
Viruses consist of DNA or RNA enclosed in a
simple protein shell known as capsid.

General properties of viruses

-​ They are very small in size, from 20-300
ηm.
-​ They contain one kind of nucleic acid (RNA
or DNA) as their genome.
-​ They are metabolically inert
Cytoplasmic Membrane
• The cytoplasmic membrane is a highly selective
permeability barrier constructed of lipids and
proteins that forms a bilayer with hydrophilic
exteriors and a hydrophobic interior.
• The major function of the cytoplasmic
membrane is to act as a permeability
barrier, preventing leakage of
cytoplasmic metabolites into the
environment.

Cell Growth
• Microbial growth involves an increase
in the number of cells. Growth of most
microorganisms occurs by the process of
binary fission.
• Microbial populations show a
characteristic type of growth pattern
called exponential growth.

Cell Wall
• Gram-negative Bacteria have only a
few layers of peptidoglycan , but gram-
positive Bacteria have several layers.
• In addition to peptidoglycan, gram-
negative Bacteria contain an outer
membrane consisting of
lipopolysaccharide (LPS), protein, and
lipoprotein.
 Low or High pH
• The acidity or alkalinity of an
environment can greatly affect microbial
growth.
• Organisms that grow best at low
pH are called acidophiles; those
that grow best at high pH are called
alkaliphiles.
• Some organisms have evolved to grow
best at low or high pH, but most
organisms grow best between pH 6 and
8. The internal pH of a cell must stay
relatively close to neutral even though the
external pH is highly acidic or basic.

The Growth Cycle

• Microorganisms show a characteristic
growth pattern when inoculated into a fresh
culture medium.

Salinity
• Some microorganisms (halophiles)
have evolved to grow best at reduced
water potential, and some (extreme
halophiles) even require high levels of
salts for growth.

Environmental Effects on
Microbial Growth

Temperature
• Temperature is a major environmental
factor controlling microbial growth. The
cardinal temperatures are the
minimum, optimum, and maximum
temperatures at which each organism
grows. Oxygen
• Aerobes require oxygen to live,
whereas anaerobes do not and may
even be killed by oxygen.
• Facultative organisms can live
with or without oxygen.
Aerotolerant anaerobes can
tolerate oxygen and grow in its
presence even though they cannot
use it.
• Microorganisms can be grouped by • Microaerophiles are aerobes that can
the temperature ranges they use oxygen only when it is present at
require. levels reduced from that in air.
 The importance of parasitology

●​ Six major tropical diseases to which WHO

pays great attention include: malaria,
schistosomiasis, filariasis, leishmaniasis,
trypanosomiasis and leprosy.
●​ Five of them are parasitic diseases except
• Special techniques are needed to grow leprosy.
aerobic and anaerobic microorganisms. ●​ All the above diseases are prevalent in
Ethiopia
Toxic Forms of Oxygen
Why were they selected?
• Several toxic forms of oxygen can be ●​ Schistosomiasis - 200,000,000 infected
formed in the cell as the result of 500,000-1,000,000 deaths/year
respiration, but enzymes are present that ●​ Malaria - 500,000,000 infected 2,500,000
can neutralize most of them. Hydrogen deaths/year
peroxide is one of those forms that can ●​ Filariasis - 250,000,000 infected
be neutralized by catalase. ●​ Trypanosomiasis - 25,000,000 infected
65,000 deaths/year
• Several toxic forms of oxygen can be ●​ Leishmaniasis - 1,200,000 infected
formed in the cell as the result of
respiration, but enzymes are present 1.3. Concepts related to parasitology
that can neutralize most of them. 1.3.1. Symbiosis
Hydrogen peroxide is one of those Any association more or less permanent is called a
forms that can be neutralized by symbiosis, with each member a symbiont.
catalase. Two different organisms live together and interact,
one partner lives in or on another one’s body.
●​ 3 types: Mutualism, Commensalism and
Parasitism
Lecture 2: Intro to Parasitology
Mutualism
1.1. Definition
●​ Permanent association between two
Parasitology (GK: para = beside Sitos = food
different organisms that life apart is
●​ The study of the parasites of man and their
impossible,
medical consequences .
●​ Two partners benefit each other,
It is a subject that researches:
●​ The mutuals are metabolically dependent
●​ the biological features of human parasites,
on one another;
●​ the relationship between the human being
●​ One cannot survive in the absence of the
and the parasites,
other.
●​ the prevention and treatment of the parasitic
diseases.
Commensalism
●​ Association of two different organisms
1.2. Scope of Parasitology
●​ One partner is benefited while the other
According to the very broad definition of
neither benefited nor injured, such as E.
parasitology, parasites should include:-
Coli and man.
●​ viruses, bacteria, fungi,
●​ protozoa and metazoa (multi-celled
Parasitism
organisms) which infect their host species.
●​ Association of two different organisms
However, for historical reasons the first three have
●​ One partner is benefited while the other is
been incorporated into the discipline of
injured, such as ascaris lumbricoides and
Microbiology.
man.
Therefore, parasitology consists of:- Protozoa
(single celled animals), Helminths (worms)
Arthropods
1.3.1. Parasite and types of parasites ●​ Heterogenetic Parasites:
Parasite:- -​ One with alteration of generations
●​ In parasitism, parasite is the benefited e.g., Coccidial parasites and
partner. Strongyloides
●​ It is an animal organism which lives in or on
the host in order to obtain nourishment and V. Based on host ranges
shelter from the host as well as does harms ●​ Euryxenous parasites:
to the host. -​ Those with a broad host range.
In another words
●​ A small organism (Parasite) has the ●​ Stenoxenous parasites:
potential to harm a larger organism (Host), -​ Those with a narrow host range;
and relies on said host for nutrients and
shelter (a Niche). Other terminology
●​ The parasite generally has a much higher ●​ Aberrant parasite:
reproductive capability compared to its host. -​ Found in locations in the host where
they normally do not occur;
Types of Parasites -​ e.g., Ascaris larvae may migrate to
Parasite can be Classified the brain

I. According to their habitat ●​ Insidental parasite:

●​ Endoparasite -​ Occurs in hosts where it does not
-​ Lives inside the body of the host normally occur;
-​ May be just under the surface or -​ e.g., Fasciola normally does not
deep in the body (Tapeworms, occur in man but is incidental if
flukes, protozoans) found in man’s liver

●​ Ectoparasite 1.3.3. Hosts and types of hosts

-​ Stays on outside surface of the host
(leeches, ticks, fleas, brood Hosts are organism which harbors the parasite.
parasites) -​ In parasitism, it is the injured partner
-​ Types of Hosts: Definitive host:-
II. Based on dependency on the host Intermediate host:
●​ Obligate Parasite
-​ Requires finding and invading the ●​ Definitive host:- What characterizes the
host to complete its life cycle primary host?
-​ Most of the parasites we will cover -​ Where sexual reproduction takes
are obligate parasites place.
-​ Normally where the adult parasites
●​ Facultative Parasite live.
-​ May become parasitic if it is given -​ Normally the larger of the hosts,
the chance but does not require a usually a vertebrate.
host. -​ Convention - (parasites which only
-​ reproduce asexually)
III. Amount of time spent -​ Specificity - frequently, a large
●​ Permanent Parasite number of host species can act as
-​ Lives entire adult life stage on or in a intermediate host and only one or a
host few can act as a definitive host
-​ Usually endoparasites (One
exception is eyelash mite) ●​ Intermediate host:-
-​ sexually immature or larval stage of
●​ Temporary Parasite a parasite
-​ Spends only a short time on a host -​ Asexual multiplication takes place
-​ Usually ectoparasites -​ may harbor many immature stages
of a parasite;
III. According to their Pathogenicity: -​ e.g., Cercaria, Redia and Sporocysts
●​ Pathogenic parasites which are all immature stages of
●​ Non-Pathogenic (commensal) Fasciola in the snail intermediate
●​ Opportunistic parasites host.
-​ Some parasites: require more than
IV. Based on their life cycle one intermediate host which are
●​ Monoxenous parasites: then designated as first, second
-​ Those with direct life cycles (i.e., intermediate,
with one host).
Other terminology
●​ Heteroxenous parasites: ●​ Paratenic or Transport Host
-​ Those with inderect life cycles -​ No development occurs but parasite
requiring an intermediate host (i.e., remains alive and infective to
involves 2 or more hosts). another host
 -​ May go dormant ●​ *Infective Route is the specific entrance
-​ May cause damage e.g., Toxoplasm through which the parasite invades the
species in cattle human body. Hookworms invade human
body by skin. Man gets infection with
●​ Accidental or Incidental Host ascarid by mouth.
-​ Parasite is in the “wrong” species. ●​ Infective Mode means how the parasite
-​ Parasite usually wanders around invades human body, such as the cercariae
and causes great damage because of the blood fluke actively penetrate the skin
it doesn’t know where to go then of a swimming man and the infective ascaris
dies. eggs are swallowed by man.
●​ Geohelminth
Types of Hosts -​ refers to the helminths which
complete their life cycles not
●​ Reservoir Host requiring the processes of the
-​ Any animal that carries a parasite development in intermediate hosts.
that can cause infections in humans. -​ They have only one host and a
Even if it is the normal host for that simple life cycle, such as ascarid,
parasite. hookworm, pinworm and etc.
-​ Related to the medical perspective ●​ Biohelminth
of parasitology -​ refers to the helminths which have to
undergo the development in
●​ Carrier host: intermediate hosts to complete their
-​ A person who harbors parasites has life cycles, such as filaria, liver fluke,
no any clinical symptom. He is an pork tapeworm and so on.
important source of infection in ●​ Alternation of Generation:
epidemiology e.g. human beings -​ In life cycles of some parasites,
harboring cyst form of E.histolytica there is the regular alternation of
sexual and asexual reproductions,
1.3.4. Host specificity eg.Plasmodium vivax
●​ The number of species the parasite can use ●​ *Trophozoite is a living stage of protozoa
as a definitive or intermediate host. when they can move, take food and
●​ Parasites show varying degrees of host reproduce. (It is usually the pathogenic
specificity stage.)
-​ A few parasites will infect only one ●​ *Cyst is the resting stage of a protozoa with
species a protective wall. It is usually the infective
-​ Most parasites will infect a few stage. Its functions are protection,
closely related species (or similar transmission and multiplication.
anatomy) Encystation
-​ Some parasites can infect a large Trophozoite(reversible)Cyst
group of animals Excystation
-​ A few parasites have little or no host
specificity 1.4. Epidemiology of parasite
-​ Epidemiology: The study of the patterns of
1.3.5. Vector and types of vectors diseases within populations
●​ Vector:-an organism (usually an arthropod) For parasites, this includes:
which transfers infective forms of a parasite -​ Host range – what can it infect?
from one host to the other. -​ Geographic range – where is it?
●​ Classification -​ Is it a zoonotic agent? Can it infect
1. Biological vectors:- humans?
2.Mechanical (Parathenic or transport) -​ Does it have a reservoir? A group of
vertebrates maintaining the parasite
1. Biological vectors:- characterized by the -​ Does it have a nidus? A small ecosystem
development of the parasite before its transfer to that possesses all the factors to maintain
another host the parasite..
-​ Propagative. E.g. Yersinia pestis in fleas
-​ Cyclopropagative E.g. Plasmodium vivax in 1.4.1. Geographic Distribution
Anopheles mosquitoes. ●​ Global distribution
-​ Cyclodevelopmental E.g.Onchocerca -​ parasite occur globally,
volvulus in black flies. -​ the majority occur in tropical regions,
Factors
2. Mechanical vector - no parasitic development of ●​ Favorable environmental
reproduction occurs conditions
●​ poverty, poor sanitation and
1.3.6. Other terminologies personal hygiene
●​ *Infective Stage : it is a stage when a
parasite can invade human body and
continue to live there. The infective stage of
ascarid is the embryonate egg.
Factors (Endemicity): -​ Sexual intercourse Blood
1. Presence of a suitable host transfusion
2. Habits of the host -​ Direct skin penetration
3. Escape from the host
4. Favorable conditions outside of host ●​ Vertical Direct Mode of Transmission: -
5. Economic and social conditions Transmission of the parasite is from the
mother to child through:
1.4.2. Transmission of parasites -​ Congenital / transplacental
Factors required:(Three key links of parasitic -​ Transmammary (breast milk
disease transmission)
1. Source of infection ●​ Indirect Mode of Transmission:-
2. Mode of transmission -​ If the parasite has complex life cycle,
3. Susceptible people -​ requires biological vectors and/or
-​ one or more intermediate hosts
1.4.2.1. source of exposure
1. primary source (infected persons, carriers, Route of Transmission
animals)
1.​ By ingesting infective stage of parasites:
1. Sources of Exposure to Parasitic Infections -​ In food, water or hands
●​ 1. Contaminated soil:- Soils polluted with contaminated with faeces, E.g. E.
human excreta is commonly responsible for histolytica, E. vermicularis, etc.
exposure to infection with geohelminthes -​ In raw or undercooked meat, e.g. T.
saginata, T. solium, T. spiralis
●​ 2. Contaminated water:- -​ In raw or undercooked fish, crab, or
●​ Water may contain water vegetation e.g. intestinal
-​ (a) viable cysts of Amoeba, flukes
flagellates etc, -​ Water containing Cyclope e.g., D.
-​ (b) cercarial stages of human blood medinensis
fluke, 2.​ Penetration of Skin When in Contact
-​ (c) Cyclops containing larva of with:
Dracunculus medinensis -​ Faecally polluted soil, e.g.,
-​ (d) fresh water fishes which are S.stercoralis, Hook worms
sources for fish tape worm, and -​ Water containing infective stages of
intestinal flukes infection the parasite E.g., Cercaria of
-​ (e) crab or cray fishes that are Schistosome species
sources for lung fluke and
-​ (f ) Water plants which are sources III. Through Insect Bite,
for Fasciolopsis buski. ●​ E.g., filarial worms, Trypanosoma sp,
-​ Plasmodium sp. etc.
C. Raw or Insufficiently cooked meat of pork, ●​ Sexual Contact, e.g., Trichomonas vaginalis
beef and fish ●​ Transmammary, e.g., S. stercoralis
-​ E.g., Trichinella spiralis, Taenia species, ●​ Inhalation of contaminated air, e.g., E.
D.latum. vermicularis, P. carnii
●​ Transplacental, e.g., T. gondii
D. Blood sucking arthropods: ●​ Kissing, e.g., Trichomonas gingivalis, T.
-​ Malaria - anopheles mosquito, tenax
-​ Leishmania - sand flies
-​ Trypanosoma - tsetse flyb 1.5 General Life Cycles of parasites
●​ Describes the cycle of development of the
E. Animals (a domestic or wild animals parasite,
harboring the parasite) ●​ This may involve
-​ e.g, 1. Dogs- the hydatid cyst caused by E. -​ Passing through a number of
granulosus developmental stages & enviroment
-​ Parasitic and non-parasitic stages.
F. Human beings: ●​ The life of a parasite can be divided into a
-​ A person his/her clothing, bedding or the number of phases:
immediate environment that he/she -​ Growth and maturation,
contaminated -​ Reproductive (sexual and asexual)
-​ Autoinfection: - e.g., S. stercoralis, E. and
vermicularis, and T. solium -​ Transmission phases.
-​ All vitally important for the
1.4.2.2. Mode of Transmission successful survival of the parasite.
Direct mode of Transmission:- classified as: ●​ Can be simple or complex depending on
how many different hosts it requires to
●​ Horizontal Direct mode of transmission: complete its cycle
Transmission is mainly effected through:- -
- Feco-oral route: most intestinal parasites
transmitted in this way.
 1.6 Parasitic Infections & Disease:
●​ Not all parasitic infections cause disease of
clinical significance.
●​ Both host and parasitic factors are involved
for the parasitic infection to cause disease
or not

1.6.1. Host Factors

1. Genetic factors, E.g. Black population who lack
Duffy antigen resist P.vivax
●​ Simple or Direct Life Cycle 2. Age,
(monoxenous) 3. Sex : e.g., T.vaginalis
-​ only one host is required to complete 4. Level of immunity: natural and acquired immunity
its cycle 5. Nutrition (malnutrition or under nutrition)
-​ the parasite often spends most of its 6. Intensity and frequency of infections
life, usually as an adult, and where it 7. Presence of co-existing disease or conditions,
reproduces which reduces immune response. e.g. Pregnancy,
-​ Transmitted from one host to HIV
another through the air, by a fomite, 8. Life style and occupation
or in contaminated food or water.
1.6.2. Parasite factors

1. Strain of the parasite and adaptation to human

host
2. Parasite load ( number of parasite )
3. Site (s) occupied in the body
4. Metabolic processes of the parasite, particularly
the nature of any waste products or toxins
produced by the parasite during its growth and
reproduction..

●​ Indirect or heteroxenous life cycles 1.6.3. How do Parasites Cause Inquiry to their
-​ requires 2 or more hosts (a vector or Host?
intermediate host ) to reproduce or ●​ Competition for the host’s nutrients
grow in - Eg. D.latum absorbs vitamin B-12, can
-​ Frequently this may involve passing cause anemia
through a number of developmental - other tapeworms absorb large amounts of
stages & Evt. proteins and sugars Use of host’s fluids -
hookworm ingests blood, can be up to 250
ml/day
●​ Destruction of host tissues
- some injure upon entry, some after
established
- eg. Swimmers itch, cercariae penetrate
and cause inflammation
- intestinal worms, after established cause
small lesions in gut, possible secondary
infection
- Entamoeba actively digest epithelial cells
in large intestine
1.5.3. Why study ●​ Tissue changes
life cycles? - may cause serious consequences to host
Control. - hyperplasia,. Eg Fasciola
Treatment. - hypertrophy,
Epidemiology. - metaplasia, change of tissue cell type to
Fundamental research another type.Eg. Paragonims (lung fluke)
- neoplasia, growth of cell to form a new
structure. Eg.Tumors

Toxins and secretions

●​ Some may cause pathogenic response,
some may inhibit immune function - eg.
Mosquito saliva
●​ Mechanical interference
- Elephantiasis (filarial worms) blocks
lymphatic system
- Tapeworms in large numbers can block
intestine
- Plasmodium can cause RBC’s to stick together
and clog capillaries

Host Immunity & Immuno – evasion of the

parasite

1.7.1. Host Responses

●​ Nonspecific immunity
-​ Macrophage endocytosis
Common for bacteria and small
protozoa
-​ Inflammation
Acute – edema and increase of
leukocytes
Subacute – monocytes and
lymphocytes present, with fibrocytes
binding parasite with collagen.
Chronic – plasma cells present and
form a granuloma
Hyperplasia – parasite causes host
to produce more cells Liver fluke
simulating enlargement of bile duct
Neoplasia (cancer) – rare parasites
have been associated with cancer,
but mechanisms are still unknown.

●​ Specific Immunity
-​ Humeral response: Formation of
antibodies or immunoglobulins (Ig)
by B cells.
-​ IgE fights helminths
-​ IgM and IgG important against
protozoans
-​ Cell mediated response: uses T-cells
Cytotoxic T cells inject invading
parasites
-​ Also release cytokines, which
promote nonspecific immunity.
(interconnected)

1.7.2. Parasite Responses

●​ Antigenic variation
-​ Change surface glycoproteins
regularly
●​ Being poorly antigenetic
-​ Don’t induce a response, or a most a
mild one
●​ Hide within host cells
-​ Host can’t kill what it can’t find
●​ Camouflage
-​ Use bits of host cells and attach to
parasite’s surface
●​ Depress host’s immune response
-​ Modulate produce of host T cell
production
 Microbiology:
Tools and Techniques
3.1 Methods of Culturing
Microorganisms: The Five I’s
• Microbiologists use five basic techniques to manipulate, grow,
examine, and characterize microorganisms in the laboratory:
inoculation, incubation, isolation, inspection, and identification
Figure 3.1
Inoculation and Isolation

• Inoculation: producing a culture

• Introduce a tiny sample (the inoculums) into a
container of nutrient medium
• Isolation: separating one species from another
• Separating a single bacterial cell from other cells and
providing it space on a nutrient surface will allow that
cell to grow in to a mound of cells (a colony).
• If formed from a single cell, the colony contains cells
from just that species.
Figure 3.2
Streak Plate Method

• Streak plate method- small droplet of

culture or sample spread over surface of
the medium with an inoculating loop
• Uses a pattern that thins out the sample and
separates the cells

Figure 3.3 a,b

Loop Dilation Method

• Loop dilation, or pour plate, method- sample

inoculated serially in to a series of liquid agar
tues to dilute the number of cells in each
successive tubes
• Tubes are then poured in to sterile Petri dishes
and allowed to solidify

Figure 3.3 c,d

Spread Plate Method

• Spread plate method- small volume of liquid, diluted sample pipette on to

surface of the medium and spread around evenly by a sterile spreading tool

Figure 3.3 e,f

Media: Providing Nutrients in the
Laboratory
• At least 500 different types
• Contained in test tubes, flasks, or Petri dishes
• Inoculated by loops, needles, pipettes, and swabs
• Sterile technique necessary
• Classification of media
• Physical state
• Chemical composition
• Functional type
Classification of Media by Physical
State
• Liquid media: water-based solutions, do not
solidify at temperatures above freezing, flow freely
when container is tilted
• Broths, milks, or infusions
• Growth seen as cloudiness or particulates
• Semisolid media: clotlike consistency at room
temperature
• Used to determine motility and to localize reactions at
a specific site
• Solid media: a firm surface on which cells can
form discrete colonies
• Liquefiable and nonliquefiable
• Useful for isolating and culturing bacteria and fungi
Figure 3.4
Classification of Media by Chemical
Content
• Synthetic media- compositions are precisely chemically
defined
• Complex (nonsynthetic) media- if even just one component is
not chemically definable
Classification of Media by Function

• General purpose media- to grow as broad a spectrum of

microbes as possible
• Usually nonsynthetic
• Contain a mixture of nutrients to support a variety of microbes
• Examples: nutrient agar and broth, brain-heart infusion, trypticase soy
agar (TSA).
Enriched Media

• Enriched media- contain complex

organic substances (for example
blood, serum, growth factors) to
support the growth of fastidious
bacteria. Examples: blood agar,
Thayer-Martin medium (chocolate
agar)
Figure 3.6
Selective and Differential Media

• Selective media- contains one or more

agents that inhibit the growth of certain
microbes but not others. Example: Mannitol
salt agar (MSA), MacConkey agar, Hektoen
enteric (HE) agar.
• Differential media- allow multiple types of
microorganisms to grow but display visible
differences among those microorganisms.
MacConkey agar can be used as a
differential medium as well.
Figure 3.7
Figure 3.8
Figure 3.9
Miscellaneous Media

• Reducing media- absorbs oxygen or slows its

penetration in the medium; used for growing
anaerobes or for determining oxygen
requirements
• Carbohydrate fermentation media- contain sugars
that can be fermented and a pH indicator; useful
for identification of microorganisms
• Transport media- used to maintain and preserve
specimens that need to be held for a period of
time
• Assay media- used to test the effectiveness of
antibiotics, disinfectants, antiseptics, etc.
• Enumeration media- used to count the numbers of
organisms in a sample.
Figure 3.10
Incubation

• Incubation: an inoculated sample is placed in

an incubator to encourage growth.
• Usually in laboratories, between 20° and 40°C.
• Can control atmospheric gases as well.
• Can visually recognize growth as cloudiness in
liquid media and colonies on solid media.
• Pure culture- growth of only a single known
species (also called axenic)
• Usually created by subculture
• Mixed culture- holds two or more identified
species
• Contaminated culture- includes unwanted
microorganisms of uncertain identity, or
contaminants.
Inspection and Identification

• Inspection and identification: Using appearance as well as metabolism

(biochemical tests) and sometimes genetic analysis or immunologic testing to
identify the organisms in a culture.

• Cultures can be maintained using stock cultures

• Once cultures are no longer being used, they must be sterilized
and destroyed properly.
3.2 The Microscope: Window on
an Invisible Realm
• Two key characteristics of microscopes:
magnification and resolving power
• Magnification
• Results when visible light waves pass through
a curved lens
• The light experiences refraction
• An image is formed by the refracted light
when an object is placed a certain distance
from the lens and is illuminated with light
• The image is enlarged to a particular degree-
the power of magnification
Figure 3.13
Principles of Light Microscopy

• Magnification- occurs
in two phases
• Objective lens- forms
the real image
• Ocular lens- forms the
virtual image
• Total power of
magnification- the
product of the power
of the objective and
the power of the
ocular
Resolution

• Resolution- the ability to distinguish two adjacent objects or

points from one another
• Also known as resolving power
• Resolving power (RP) = Wavelength of light in nm
2 x Numerical aperture of
objective lens
• Resolution distance= 0.61 x wavelength of light in nm
Numerical aperture of objective
lens
• Shorter wavelengths provide a better resolution
• Numerical aperture- describes the relative efficiency of a lens in
bending light rays
• Oil immersion lenses increase the numerical aperture
Figure 3.15
Figure 3.16
Magnification and Resolution

• Increased magnification decreases the resolution

• Adjusting the amount of light entering

the condenser using an adjustable iris
diaphragm or using special dyes help
increase resolution at higher
magnifications
Figure 3.17
Variations on the Optical
Microscope
• Visible light microscopes- optical microscopes that use visible
light. Described by their field.
• Four types: bright-field, dark-field, phase-contrast, and interference

• Other light microscopes include fluorescence microscopes and

confocal microscopes
Bright-Field Microscopy

• Most widely used

• Forms its image when light is transmitted through the specimen
• The specimen produces an image that is darker than the
surrounding illuminated field
• Can be used with live, unstained and preserved, stain
specimens
Dark-Field Microscopy

• A bright-field microscope can be adapted to a

dark-field microscope by adding a stop to the
condenser
• The stop blocks all light from entering the objective
lens except for peripheral light
• The specimen produces an image that is brightly
illuminated against a dark field
• Effective for visualizing living cells that would be
distorted by drying or heat or that can’t be
stained with usual methods
• Does not allow for visualization of fine internal
details of cells
Phase-Contrast Microscopy

• Transforms subtle changes in light waves passing through a

specimen into differences in light intensity
• Allows differentiation of internal components of live, unstained
cells
• Useful for viewing intracellular structures such as bacterial
spores, granules, and organelles
Figure 3.18
Interference Microscopy

• Interference Microscopy
• Uses a differential-interference contrast (DIC) microscope
• Allows for detailed view of live, unstained specimens
• Includes two prisms that add contrasting colors to the image
• The image is colorful and three-dimensional
Figure 3.19
Fluorescence Microscopy

• Includes a UV radiation source and a filter that protects the

viewer’s eyes
• Used with dyes that show fluorescence under UV rays
• Forms a colored image against a black field
• Used in diagnosing infections caused by specific bacteria,
protozoans, and viruses using fluorescent antibodies
Figure 3.20
Confocal Microscopy

• Allows for viewing cells at higher magnifications using a laser

beam of light to scan various depths in the specimen
• Most often used on fluorescently stained specimens
Figure 3.21
Electron Microscopy

• Originally developed for studying nonbiological

materials
• Biologists began using it in the early 1930s
• Forms an image with a beam of electrons
• Electrons travel in wavelike patterns 1,000 times shorter
than visible light waves
• This increases the resolving power tremendously
• Magnification can be extremely high (between
5,000X and 1,000,000X for biological specimens)
• Allows scientists to view the finest structure of cells
• Two forms: transmission electron microscope (TEM)
and scanning electron microscope (SEM)
TEM

• Often used to view structures of cells and viruses

• Electrons are transmitted through the specimen
• The specimen must be very thin (20-100 nm thick) and stained
to increase image contrast
• Dark areas of a TEM image represent thicker or denser parts
Figure 3.22
SEM

• Creates an extremely detailed three-dimensional view of all

kinds of objects
• Electrons bombard the surface of a whole metal-coated
specimen
• Electrons deflected from the surface are picked up by a
sophisticated detector
• The electron pattern is displayed as an image on a television
screen
• Contours of specimens resolved with SEM are very revealing
and surprising
Figure 3.23
Preparing Specimens for Optical
Microscopes
• Generally prepared by mounting a sample on a glass slide
• How the slide is prepared depends on
• The condition of the specimen (living or preserved)
• The aims of the examiner (to observe overall structure, identify
microorganisms, or see movement)
• The type of microscopy available
Living Preparations

• Wet mounts or hanging drop mounts

• Wet mount:
• Cells suspended in fluid, a drop or two of the
culture is then placed on a slide and overlaid with
a cover glass
• Cover glass can damage larger cells and might
dry or contaminate the observer’s fingers
• Hanging drop mount:
• Uses a depression slide, Vaseline, and coverslip
• The sample is suspended from the coverslip
Figure 3.24
Fixed, Stained Smears

• Smear technique developed by Robert Koch

• Spread a thin film made from a liquid suspension
of cells and air-drying it
• Heat the dried smear by a process called heat
fixation
• Some cells are fixed using chemicals
• Staining creates contrast and allows features
of the cells to stand out
• Applies colored chemicals to specimens
• Dyes become affixed to the cells through a
chemical reaction
• Dyes are classified as basic (cationic) dyes, or
acidic (anionic) dyes.
Positive and Negative Staining

• Positive staining: the dye sticks to the

specimen to give it color
• Negative staining: The dye does not stick
to the specimen, instead settles around its
boundaries, creating a silhouette.
• Nigrosin and India ink commonly used
• Heat fixation not required, so there is less
shrinkage or distortion of cells
• Also used to accentuate the capsule
surrounding certain bacteria and yeasts
Simple Stains

• Require only a single dye

• Examples include malachite green, crystal violet, basic fuchsin, and
safranin
• All cells appear the same color but can reveal shape, size, and
arrangement
Differential Stains

• Use two differently colored dyes, the primary dye and the
counterstain
• Distinguishes between cell types or parts
• Examples include Gram, acid-fast, and endospore stains
Gram Staining

• The most universal diagnostic staining technique for bacteria

• Differentiation of microbes as gram positive(purple) or gram
negative (red)
Acid-Fast Staining

• Important diagnostic stain

• Differentiates acid-fast bacteria (pink) from non-acid-fast
bacteria (blue)
• Important in medical microbiology
Endospore Stain

• Dye is forced by heat into resistant bodies called spores or

endospores
• Distinguishes between the stores and the cells they come from
(the vegetative cells)
• Significant in medical microbiology
Special Stains

• Used to emphasize certain cell parts that aren’t revealed by

conventional staining methods
• Examples: capsule staining, flagellar staining
Figure 3.25
Classification of
Helminths
 PROTOZOA HELMINTHS
CLASSIFICATION OF PARASITES
Unicellular Multicellular
Single cell for all functions Specialized cells
Intestinal ,blood & tissue. Intestinal & tissue.
1:Aoebae: move by Round worms (Nematodes):
pseudopodia. - elongated, cylindrical,
2:Flagellates: move by flagella. unsegmented.
3:Ciliates: move by cilia Flat worms :
4:Apicomplexa(Sporozoa) - Trematodes: leaf-like,
tissue parasites unsegmented.
- Cestodes: tape-like,
segmented.
Nematodes: Location in the human body

• Intestinal nematodes
• Tissue nematodes
 1-Nematodes : General features

1. Elongated worm, cylindrical, un-segmented and

tapering at both ends.
2. Variable in size, measure <1 cm to about
100cm.
3. Sex separate and male is smaller than female
Nematodes: common intestinal infections
Common intestinal nematode infections:
1. Enterobius (Oxyuris) vermicularis
(Pinworm,seatworm,threadworm)

2. Trichuris trichiura (whipworm)

3. Ascaris lumbricoides (roundworm)
4. Ancylostoma duodenale & Necator americanus
(hookworms)

5. Strongyloides stercoralis
6. :
 1-Enterobius vermicularis (THREAD WORM)

(Common names :Pin worm, seat worm, (

 Found all over the world but more common in
temperate regions and in Saudi Arabia.
 Children are more often involved than adults , it
tends to occur in groups living together such as
families , army camps or nursery.
 Adult worms are mainly located in lumen of cecum
and the female migrate to rectum to deposits her
eggs on the anus and perianal skin.
 .Adult worm can be seen by naked eye as white
thread ± 1cm.
 Male is smaller than female ± 0.5cm, with coiled end.
 .

1-Enterobius vermicularis (THREAD WORM)
(Common names :Pin worm, seat worm, (
 Found all over the world but more common in
temperate regions.
 Children are more often evolved than adults , it
tends to occur in groups living together such as
families , army camps or nursery.
 Adult worms are mainly located in lumen of cecum
and the female migrate to rectum to deposits her
eggs on perianal skin.
 Direct human to human infection occurs mainly by
swallowing the eggs .In addition ,autoinfection
occurs by contamination of the fingers.
 It can be seen by naked eye as white thread ± 1cm.
 Male is smaller than female ± 0.5cm, with coiled end.
Enterobius vermicularis (pin worm)
 Clinical manifestations of pin worm
 The most common symptom is perianal itching, also
known as pruritus ani ,
 which can be very troublesome and occurs more often
during the night, persistent itching may lead to
inflammation and secondary bacterial infection of the
perianal region . also adult worm can lodged in the
lumen of appendix cause appendicitis.
 Infected children may suffer from emotional
disturbance ,insomnia ,anorexia , loss of weight and
loss of concentration and enuresis.

 Ectopic infection occurs in infected adult female

when invade vulva and vagina result in
valvo-vagintis, salpingitis.
 Enterobius vermicularis (Oxyuris)
DIAGNOSIS :
Unlike other intestinal Nematodes, the eggs
are not usually found in feces .The best
method is to look for them around the anus
by taking an anal swab or by using
CELLULOSE ADHESIVE TAPE, the
examination should be done before
defecation or bathing.
Treatment
ِِAlbandazole , Mebendazole
for whole family
Enterobius vermicularis (pin worm)
 Ascaris lumbricoides
(roundworm)
The commonest human helminthes infection all
over the world.
The large round worm which is normally located
in the small intestine.
 Found in jejunum and upper part of ileum.

 Female ± 20 cm longer than male ± 10 cm

 Feed on semi digested food.

Life cycle of Ascais Lumbricoides
Infection starts when man ingest an Embryonated
egg contaminated with food or water(soil), then
this embryonated egg become a Larva in the
duodenum, and penetrate the wall of the
duodenum , enter the blood stream to the heart ,
liver and enter the pulmonary circulation and stay
in the alveoli ,where it grow for three weeks then
Larva passes from respiratory system to be
coughed up ,swallowed ,returned back to the small
intestine where it mature to adults male &female
,fertilization take place producing eggs which pass in
stool as Fertilized eggs or unfertilized eggs ,only
fertilized eggs can be survive in the( soil) for 2
weeks to become an Embryonated egg ready to
infect human with contaminated food.
a
Ascaris lumbricoides life cycle
 Pathogenicity
 1-Migrating LARVA :
 Ascaris pneumonia , some times LARVA reach
aberrant sites like brain ,heart or spinal cord
can cause unusual disturbance.
 2-Adult WORM:
 The worm consumes proteins and vitamins
from host’s diet and leads to malnutrition.
 Can cause intussusception, intestinal ulcers
and in massive infection can cause intestinal
obstruction.
 Ascaris lumbricoides
(roundworm)
Pathology:
 1-Adult worm:
Light infection : asymptomatic.
Heavy infection : intestinal obstruction
Migrating adult : to bile duct -jaundice
 2-Larvae: Loeffler`s syndrome
Pneumonitis and bronchospasm, cough with
bloody sputum
Eosinophilia, urticaria
 Ascaris lumbricoides
(roundworm)

Loeffler`s syndrome: Larvae in lung

pneumonia ,cough ,bloody sputum
 Ascaris lumbricoides
(roundworm)

Diagnosis:
-eggs (fertilized) or
unfertilized eggs in
stool.
-larvae in sputum.
-adult may pass with
stool.

Treatment: Albendazole , Mebendazole

 Trichuris trichiura (whipworm)
World wide ,common in poor
sanitation.
 It coexists with Ascaris
because of similar
requirement.
 Adult live in large intestine
especially caecum and
appendix –in heavy
infection the whole length of
large intestine affected.
 Male and female worm have
narrow anterior portion
penetrate the intestinal
mucosa
Trichuris trichiura (Whipworm)

In the soil the unembryonated eggs(diagnostic)

Become emberyonated(infective)
• Life cycle and transmission —
The life cycle for trichuriasis begins with passage of
unembryonated eggs(diagnostic stage) in
the stool . In the soil, the eggs become embryonated and
become infective in 15 to 30 days. After ingestion of
contaminated food or water with soil contains
emberyonated eggs(infective stage),
in the small intestine the larvae will hatch and become
mature into adult worms, which become established in the
cecum and ascending colon after two to three months.
• The adults measure approximately 4 cm in length. The
females begin to produce eggs 60 to 70 days after infection
and shed 3000 to 20,000 eggs per day. The life span of the
adults is one to three years
 Trichuris trichiura (Whipworm)
clinical finding
 light infection : asymptomatic
 heavy infection :abdominal pain ,bloody
diarrhea. Rectal prolapse in children is a
common complication.
-
-Diagnosis: egg in stool characterized by its barrel shape
with mucoid plugs at each pole
Unembryonated eggs

Treatment :Albendazole.
 Hook worms
Ancylostoma dudenale &Necator americanus

 A common cause of anemia.

 Found in small intestine mainly jejunum.
 Its buccal capsule (mouth) lined with hard
hooks, triangular cutting plates and
anticoagulant glands.

 Buccal cavity attached to intestinal mucosa

 Hook worms
Ancylostoma dudenale &Necator americanus

Infection occurs by
penetration of the larva
to the human skin,
In the soil
eggs(diagnostic stage)
become larva (infective
stage)
 Hook worms
Pathology& clinical picture:
- larvae:
At the site of entry of larvae intense itching(ground itch)
and dermatitis.
Migration phase:
cough with bloody sputum
pneumonitis and bronchitis but less sever than
Ascaris ,eosinophilia urticaria.
- Adult worm:
 low worm burden (INFECTION): no symptoms.
 Moderate to heavy burden:
•Epigastric pain, vomiting , hemorrhagic enteritis.
•Protein loss: hypo-proteinaemia edema.
•Anemia: due to withdrawal of blood by parasites and
hemorrhage from punctured sites lead to sever anemia =
microcytic- hypo chromic anemia .
•Iron deficiency anemia.
 Hook worms
Diagnosis and treatment

 Diagnosis:
 -Eggs in stools.;
-occult blood (+)

Treatment: Albendazol, Mebendazole

 Strongyloides stercoralis

 Widely distributed in tropical area at Asia,

Africa & South America .
 fatal dissemination in
immuno-compromised host.
 It is smallest pathogenic nematodes
± 2.5mm.
 adult live in mucous membrane of duodenum
jejunum rarely mucous membrane of
bronchus.
 AUTOINFECTION IS VERY IMPORTANT
CRITERIA .
Strongyloides stercoralis

LIFE CYCLE
Strongyloids Stercoralis
 1-Rabditiform larva (diagnostic stage) are excreted in the
stool to the soil , to become adult male and female, where
fertilization take place to produce eggs.
 2- Larva hatch from the eggs in the soil and become
Filariform larva (infective stage) .
 3-Infection take place by penetration of the Filariform larva to
human skin ,which enter the circulation to the lung trachea
 swallowed and reach the small intestine to become
adult male and female ,starts to produce eggs ,which become
Rabditiform larva and excreted in the stool to start another
cycle in the soil.
 INTERNAL AUTOINFECTION in immunocompromised
patients ,Rabditiform larva in the intestine can be transferred
to become filariform larva and penetrate the intestinal
mucosa and peri-anal skin and produce infection.
 Strongyloides stercoralis:
Pathology and clinical picture:

 Cutaneous little reaction on penetration.

sever dermatitis at perianal region in
case of external autoinfection.
 Migration :same as hook worms .
 Intestinal: inflammation of upper intestinal mucosa,
diarrhea, upper abdominal pain colicky in nature.
 Disseminated strongyloidiasis : in patient with
immunodeficiency ,uncontrolled diarrhea , necrosis
,perforation--peritonitis--death.
 Strongyloides stercoralis

Diagnosis:
rhabditiform larvae
diagnostic stage in:
-Stool examination
-Duodenal aspirate

Treatment :
Albandazole, Mebendazole
 2-Tapeworm (Cestodes) Infections
LAB.

TRANSMISSION CLINICAL DIAGNOSIS

TAPEWORM LOCATION LOCATION
DISEASE OF INFECTION OF ADULT OF LARVA
PICTURE
IN HUMANS IN
HUMANS

Taenia taeniasis ingestion of larva Small not vague eggs or

saginata in undercooked Intestine present digestive proglottids
beef in stools
disturbance
Taenia solium- taeniasis ingestion of larva Small not s
vague digestive eggs or
in undercooked Intestine present disturbances proglottids
pork in stools
ADULT
Taenia not present sub- depending X-
solium- Cysticercosis ingestion of egg (except in cutaneous on ray,CT,MRI
autoinfection muscles locality: Serology
LARVA
: brain,eyes from none
(cysticercus ,small to
cellulosae) intestine) epilepsy
Hymenolepis hymenolepiais ingestion of egg Small Intestinal Enteritis eggs in
nana Intestine Villi diarrhoea stools

Echinochoccu hydatid ingestion of egg not present Liver, depending X-ray,CT,US

lungs, Serology
s disease on Hydatid sand
Bones etc
granulosus locality
Taenia saginata
Human(definitive host harboring adults) infected by eating
Under cooked beef contains cysticercus in the muscle of
the cattle(intermediate host).
 Taenia saginata
 Is an obligatory parasite of man ,the adult worm live in the
SMALL INTESTINE .
 CATTLE become infected by ingesting grass contaminated
with eggs or gravid segments which passed from human
faeces. In the cattle the onchosphere hatches out go to
circulation and transformed to cysticercus stage in the muscle
known as CYSTICERCUS BOVIS.
 Man become infected by eating undercooked or improperly
cooked beef , the adult worm lives in small intestine of
man passing eggs and gravid proglottids to the
environment.
 The majority of cases are Asymptomatic ,some
patients have vague intestinal discomfort ,vomiting
and diarrhoea.
T.Saginata infection is
usually
asymptomatic ,but in
heavy infection often
result in weight loss
,dizziness ,abdominal
pain, diarrhea and
loss of appetite.

Diagnosis :
detection of eggs in
stool or gravid
segment.
Life cycle of T.Solium
HYMENOLEPIS NANA
 Life cycle of Echinococcus granulosus

1-The adult worm located in the small intestine of the dogs

(definitive host).
2-Eggs and gravid segments are then discharged in the feces of
the infected dogs having Onchosphere.
3- Eggs and gravid segments contain onchosphere are
ingested by various animals e.g sheep goats and
accidentally man.
These onchosphere hatches in the duodenum and penetrate
the liver
the circulation and reach various organs mainly
and the brain causing HYDATID CYST.
Echinococcus granulousus
 Liver hydatid cyst

Diagnosis :
1- radiological examination.
2- immunological tests.

Treatment:
Intestinal stages: Praziquantel
Tissue stages ( Hydatid , cysticersosis):
Depends on clinical condition : Surgical and/or
Albendazole
Hydatid cyst
Microbial control
Microbial control
What is control ?
 Restricting the growth and normal activities of
microorganisms.
Microbial control
Why control ?
 To prevent transmission of diseases.
 To prevent contamination at various levels.
 To prevent spoilage of materials.
Microbial control
How control ?
 By killing the microorganisms (cidal)
 By inhibiting the microorganisms (static)
 By removing the microorganisms
Microbial control
Mechanism of antimicrobial action
 Affecting the cell wall
 Affecting the cell membrane
 Interfering with protein structure and function
 Affecting nucleic acids
Microbial control
Factors affecting antimicrobial activities
 Type and state of microorganisms
 Initial microbial load
 Concentration or dose of agent
 Time of exposure
 Environmental conditions
 Microbial control - Practical Concern

 Does the application require sterilization ?

 Is the item to be reused ?
 Can the item withstand heat, pressure, radiation or
chemical ?
 Will the agent penetrate to the necessary extent ?
 Is the method cost and labor efficient and is it safe ?
 Microbial control - Some useful terms

 Sterilization :killing or removal of all life form i.e.

free from life.
 Disinfection :surface removal of organisms.
 Sanitation :reduction in number of organism.
 Antiseptic :agent that prevent sepsis.
 Pasteurization :killing of organism without affecting
quality of material.
 Kinetics of heat sterilization
 Death from heating is an exponential (first order)
function and occurs more rapidly as the temp raised.
 Thus if we practically wants to sterilize a microbial
population , it will take longer at low temp than at
higher temp.
 It is thus required to adjust time and temp to achieve
sterilization for each specific set of conditions.
 The nature of heat is also important – moist heat is
more penetrating than dry heat.
Kinetics of heat sterilization
 Kinetics of heat sterilization
Death rate-Decimal reduction time (D-value)
 It is the time in minute to kill 90% of the cells in a population.
OR
 It is the time to reduce the population by one decimal
Death rate- Thermal Death Point (TDP)
 Lowest temperature required to kill all the microorganisms in
the liquid suspension in 10 min.
Death rate- Thermal Death Time (TDT)
 Minimum time required to kill all microorganisms in liquid
suspension at given temperature
 Physical control

 Heat (low temp, high temp)

 Desiccation
 Osmotic pressure
 Radiation(ionizing & non ionizing)
 Filtration(porcelain, barkfeld, sintered glass)
 High temperature
Moist heat Dry heat
o o
 Boiling (100 C)  Incineration(>500 C)
o o
 Tyndalization (100 C,3days)  Hot air oven (160 C,2hrs.
o
 Autoclave or 170 C,1 h.)
o o
(15 lbs/sq.in.,121 C-20min.)  Flaming (>500 C)
 Pasteurization(LTHT-
o
63 C,30min.,HTST-
o
72 C,15sec.)
 Moist vs Dry heat

o
Temp( C) Time(min)
Moist heat 121 15
125 10
134 3
Dry heat 121 600
140 180
160 120
170 60
Autoclave
 Heat
Low temp.(Static) High temp.(Cidal)
o o
 Freezing (below 4 C)  Moist heat (>100 C)
o o
 Chilling (4-6 C)  Dry heat (>100 C)
 Desiccation

Control by drying - vacuum, heat etc.

Susceptibility to dessication varies widely:
 Neisseria gonnorrhea: Only survives about one hour.
 Mycobacterium tuberculosis: May survive several months.
 Viruses are fairly resistant to dessication.
 Clostridium spp. and Bacillus spp.: May survive decades.
 Filtration
 Depth filters-particulate material (porcelain, glass,
earth, asbestos)
 Membrane filters (cellulose acetate, polycarbonate,
etc.) different pore sizes; 0.2 or 0.4 micrometer
remove bacteria
 HEPA (high-efficiency particulate air) filters used for
hospitals, laminar flow hoods in laboratories, being
marketed or home use to reduce allergens
 Filtration
Laminar air flow Membrane filtration
 Osmotic pressure

 High sugar and salt conc. Can be used to restrict

the growth (Bacteriostatic).
 Many microbes can tolerate sugar conc. Up to 10%
(saccharophilic or saccharolytic).
 Some can tolerate salt conc. Up to 30%
(halophiles).
Radiation
 Radiation
Radiation: Three types of radiation kill microbes:
1. Ionizing Radiation: Gamma rays, X rays, electron
beams, or higher energy rays. Have short wavelengths (less
than 1 nanometer).
Dislodge electrons from atoms and form ions.
Cause mutations in DNA and produce peroxides.
Used to sterilize pharmaceuticals and disposable medical
supplies. Food industry is interested in using ionizing
radiation.
Disadvantages: Penetrates human tissues. May cause
genetic mutations in humans.
 Radiation
2. Ultraviolet light (Non ionizing Radiation): Wavelength is
longer than 1 nanometer. Damages DNA by producing thymine
dimers, which cause mutations.
Used to disinfect operating rooms, nurseries, cafeterias.
Disadvantages: Damages skin, eyes. Doesn’t penetrate
paper, glass, and cloth.

3. Microwave Radiation: Wavelength ranges from 1 mm to 1m

Heat is absorbed by water molecules.
May kill vegetative cells in moist foods.
Bacterial endospores, which do not contain water, are not
damaged by microwave radiation.
Radiation
Chemical Agents
 Phenolics
 Alcohols
 Halogens
 Heavy metals
 Quaternary Ammonium Compounds
 Aldehydes
 Sterilizing Gases
Chemical Agents: Phenolics
 Aromatic organic compounds with attached -OH
 Denature protein & disrupt membranes
 Phenol, orthocresol, orthophenylphenol,
hexachlorophene
 Commonly used as disinfectants (e.g. “Lysol”); are
tuberculocidal, effective in presence of organic
matter, remain on surfaces long after application
 Disagreeable odor & skin irritation; hexachlorophene
once used as an antiseptic but its use is limited as it
causes brain damage
Chemical Agents: Alcohols
 Ethanol; isopropanol; used at concentrations
between 70 – 95%
 Denature proteins; disrupt membranes
 Kills vegetative cells of bacteria & fungi but not
spores
 Used in disinfecting surfaces; thermometers;
“ethanol-flaming” technique used to sterilize glass
plate spreaders or dissecting instruments at the lab
bench
Chemical Agents: Halogens
 Act as oxidizing agents; oxidize proteins &
other cellular components
 Chlorine compounds
 Used in disinfecting municipal water supplies (as
sodium hypochlorite, calcium hypochlorite, or
chlorine gas)
 Sodium Hypochlorite (Chlorine Bleach) used at 10
- 20% dilution as bench top disinfectant
 Halazone tablets (parasulfone dichloro amido
benzoic acid) used by campers to disinfect water
for drinking
Chemical Agents: Halogens
 Iodine Compounds
 Tincture of iodine (iodine solution in alcohol)
 Potassium iodide in aqueous solution
 Iodophors: Iodine complexed to an organic
carrier; e.g. Wescodyne, Betadyne
 Used as antiseptics for cleansing skin surfaces and
wounds
Chemical Agents: Heavy Metals
 Mercury, silver, zinc, arsenic, copper ions
 Form precipitates with cell proteins
 At one time were frequently used medically as
antiseptics but much of their use has been replaced
by less toxic alternatives
 Examples: 1% silver nitrate was used as opthalmic
drops in newborn infants to prevent gonorrhea; has
been replaced by erythromycin or other antibiotics;
copper sulfate used as algicide in swimming pools
Chemical Agents: Quaternary
Ammonium Compounds
 Quaternary ammonium compounds are
cationic detergents
 Amphipathic molecules that act as
emulsifying agents
 Denature proteins and disrupt membranes
 Used as disinfectants and skin antiseptics
 Examples: cetylpyridinium chloride,
benzalkonium chloride
Chemical Agents: Aldehydes
 Formaldehyde and gluteraldehyde
 React chemically with nucleic acid and protein,
inactivating them
 Aqueous solutions can be used as disinfectants
Chemical Agents:Sterilizing Gases
 Ethylene oxide (EtO)
 Used to sterilize heat-sensitive equipment and
plastic ware
 Explosive; supplied as a 10 – 20% mixture with
either CO2 or dichlorofluoromethane
 Its use requires a special EtO sterilizer to carefully
control sterilization conditions as well as extensive
ventilation after sterilation because of toxicity of
EtO
 Much of the commercial use of EtO (for example,
plastic petri dishes) has in recent years been
replaced by gamma irradiation
Chemical Agents:Sterilizing Gases
 Betapropiolactone (BPL)
 In its liquid form has been used to sterilize
vaccines and sera
 Decomposes after several hours and is not as
difficult to eliminate as EtO, but it doesn’t
penetrate as well as EtO and may also be
carcinogenic
 Has not been used as extensively as EtO
 Vapor-phase hydrogen peroxide
 Has been used recently to decontaminate
biological safety cabinets