Topic:

o Stages in normal growth curve

o Microbial genetics
o Prokaryotic transcription & translation
o Conjugation
o Mutation and its causes
o Mechanism of drug resistance
o Pathogenesis
o Gateway to infection
o Resident Flora
o Mechanisms of Invasion
o Classic stages of clinical infection
o Sterilization and Disinfection
Microbiology is the study of microscopic organisms, such as bacteria, viruses, archaea, fungi
and protozoa
 Stages in Normal Growth curve
There are four distinct phases of the growth curve
i. Lag phase
ii. Exponential phase (log/ logarithm)
iii. Stationary phase
iv. Death/ Decline phase
The initial phase is the lag phase where bacteria are metabolically active but not dividing
The death phase is characterized by an exponential decrease in the number of living cells
  Microbial Genetics
Microbial genetics is a subject area within microbiology and genetic engineering
 Microbial genetics studies microorganisms for different purposes. The microorganisms that are
observed are bacteria, and archaea. Some fungi and protozoa are also subjects used to study in
this field
Discovery of Microbial genetics had its origins in the 1940s and 1950s, as researchers such as
George Beadle and Ed Tatum developed biochemical genetics through studying mutants of
Neurospora; Joshua Lederberg isolated mutants in Escherichia coli and discovered F-factor-
mediated conjugation
Composition of Microbial genomes encompass all chromosomal and extra
chromosomal genetic material. The study of genomes as an entity as opposed to
individual genetic components is referred to as genomics
 Microbial genomes are widely variable and reflect the enormous diversity of bacteria, archaea
and lower eukaryotes.
Applications of Microbial genetics plays a crucial role in field of molecular and cell biology
and profound applications in medicine, food, agriculture and pharmaceutical industries. It also
involves the transmission of hereditary characters in microbes.

 Prokaryotic transcription & translation

The mechanism by which the sequence of nucleotides in a gene determines the sequence of
amino acids in a protein occurs in the following steps:
1. Transcription
2. Translation
3. Post – translational modification
Transcription the process by which a sRNA is formed by RNA- polymerase using DNA as a
template, this RNA is called m RNA. mRNA has a nucleotide sequence complementary to the
template strand in the DNA.
Translation is the process of translating the sequence of a messenger RNA (mRNA) molecule to
a sequence of amino acids during protein synthesis. The genetic code describes the relationship
between the sequence of base pairs in a gene and the corresponding amino acid sequence that it
encodes
Prokaryotic translation:

Prokaryotic transcription and translation:

 Conjugation
The temporary joining together of two bacterial cells to transfer genetic material via the plasmid
(either as solitary or as part of a chromosome) from the donor cell to the recipient cell
Principle
The process of bacterial conjugation is based on the principle that the plasmid or any other
genetic material is transferred from the donor cell to the recipient cell through close physical
contact.
• Of all the conjugative plasmids, the F (fertility) plasmid of E. coli was the first discovered and
is one of the best-studied
• The F plasmid is present in one or two copies per cell and is very large (about 100 kilobases).
E. coli harboring the F plasmid are referred to as donor (F+ or male) cells and E. coli lacking the
F plasmid are referred to as recipient (F– or female) cells. Only donor cells are capable of
transferring the F plasmid to recipient cells.
• For transfer of the F plasmid from donor to recipient, intimate contact between cells, resulting
in mating-pair formation, is required
• The transfer of genetic material is then brought by membrane fusion of the two cells by the
action of different enzymes
• Following the membrane fusion, the replication of donor DNA occurs and is transferred into
the recipient cell.
Steps of Bacterial Conjugation
Examples of Bacterial Conjugation
Agrobacterium tumefactions causes’ crown gall tumor in plants by transferring the T DNA
element, a part of the Ti (tumor-inducing) plasmid present in this bacterium, into a plant cell
where the T element becomes incorporated into the plant cell’s genome
  Mutation
Mutation occurs when a DNA gene is damaged or change in such a way as to alter the genetic
message carried by that gene. A Mutagen is an agent of substance that can bring about a
permanent alteration to the physical composition of a DNA gene such that the genetic message is
changed
• Once the gene has been damaged or changed the mRNA transcribed from that gene will now
carry an altered message
The causes of mutations
• Mutations happen for several reasons
1. DNA fails to copy accurately
Most of the mutations that we think matter to evolution are "naturally-occurring." For
example, when a cell divides, it makes a copy of its DNA — and sometimes the copy is not
quite perfect. That small difference from the original DNA sequence is a mutation

2. External influences can create mutations

Mutations can also be caused by exposure to specific chemicals or radiation. These agents cause
the DNA to break down. This is not necessarily unnatural — even in the most isolated and
pristine environments, DNA breaks down. Nevertheless, when the cell repairs the DNA, it might
not do a perfect job of the repair. So the cell would end up with DNA slightly different than the
original DNA and hence, a mutation
Ionizing radiations: such as X-rays, gamma rays and alpha particles cause DNA breakage and
other damages. The most common lab sources include cobalt60 and cesium-137. Ionizing
radiations cause breaks in polysugar phosphate backbone of DNA and, thus, causing
chromosomal mutations such as break, deletion, addition, inversion and translocation.
Ultraviolet radiations: with wavelength above 260 nm are absorbed strongly by bases,
producing pyrimidine dimers, which can cause error in replication if left uncorrected
 Mechanism of Drug Resistance
These are the following main mechanisms by which microorganism’s exhibit resistance to
antimicrobials are
  Pathogenesis
Pathogenesis is the progression of processes of cellular lineage, maturation, and
migration, and eventual morphogenesis of both individual cells and their architecture in
forming a tissue or organ.
Occurrence of Pathogenesis:
Pathogenesis is the process by which an infection leads to disease include:
 Implantation of virus at the portal of entry.
 Local replication.
 Spread to target organs (disease sites),
 Spread to sites of shedding of virus into the environment.
Stages of Pathogenesis:
To cause disease, a pathogen must successfully achieve four steps or stages of pathogenesis:
exposure (contact), adhesion (colonization), invasion, and infection. The pathogen must be able
to gain entry to the host, travel to the location where it can establish ab infection, evade or
overcome the hosts immune response, and cause damage (i.e., disease) to the host. In my cases,
the cycle is completed when the pathogen exists the host and is transmitted to a new host.
Exposure:
 An encounter with a potential pathogen is known as exposure or contact. The food we eat and
the object we handle are all ways that we can come into contact with potential pathogens. Yet,
not all contacts result in infection and disease. For a pathogen to cause disease, it needs to be
able to gain access into host tissue. An anatomic site through which pathogens can pass into host
tissue is called portal of entry.
Adhesion:
Following the initial exposure, the pathogen adheres at the portal of entry. The term adhesion
refers to the capability of pathogenic microbes to attach to the cells of the body using adhesion
factors, and different pathogens use various mechanisms to adhere to the cells of host tissue.
Molecules (either proteins or carbohydrates) called adhesions are found on the
surface on the surface of certain pathogens and bind to specific receptors.
Invasion:
Once adhesion is successful, invasion can proceed. Invasion involves the dissemination of a
pathogen throughout local tissue or a body. Pathogens may produce exoenzymes or toxins,
which serve as virulence factors that allow them to colonize and damage host tissues as they
spread deeper into the body. Pathogens may also produce virulence factors determine the degree
of tissue damage the occurs. A pathogens specific virulence factors determine the degree of
tissue damage that occurs. Figure 5 shows the invasion of H. pylori into the tissues of the
stomach, causing damage a it progress.
Infection:
Following invasion, successful multiplication of the pathogen leads to infection. Infection can be
described as local, focal, or a systemic, depending on a extent of the infection. A local infection
is confined to a small area of the body, typically near the portal pf entry. For example, a hair
follicle infected by Staphylococcus aureus infection may result in a boil around the site of
infection, but the bacterium is largely contained to this small location.
Respond to pathogenesis:
The immune system responds to antigens by producing cells that directly attack the pathogen, or
by producing special proteins called antibodies. Antibodies attach to an antigen and attract cells
that will engulf and destroy the pathogen. The main cells of the immune system are lymphocytes
known as B cells and T cells.
Example of pathogenesis:
For example, bacterial pathogenesis is the mechanism by which bacteria cause infectious illness.
Most diseases are caused by multiple processes. For example, certain cancers arise from
dysfunction of the immune system (skin tumors and lymphoma after a renal transplant, which
requires immunosuppression).
 Gateway to infection
What is infection?
 An infection is the colonization of a host by microbial species. Infecting Microbes seek to
use the host resources to reproduce,often resulating in disease.Colloquially, infections are
usually considerd to be caused by microscopic organisms like viruses,prions,bacteria,and
viroids,though larger organisms like macro parasites and fungi can also infect.
Basic of classification of infections:
 Infection are classified in multiple ways. They are classified by the causative agent as
well as by the constellation of symptoms,and medical signs that are produced.
 An infection that produces symptoms is an apparent infection. An infection that is active,
but does not produce noticeable symptoms,may be called in apparent,silent,or subclinical.
 An infection that is inactive or dormant is called a latent infection.
What Causes an infection Disease?

 Infection is caused by microorganism

 The microorganism may be a bacteria,a virus,a parasite or
a fungus.
OVERVIEW:
Types of Microorganisms
Principles of infection:
 Transmission
 Host resistance
 Virulence and pathogenicity
 Control of transmission and infection
Development of the infection
 Onset and course
 Clinical signs and symptoms
 Diagnostic tests
 Antimicrobial drugs
Example of the infection: Influenza
Conditions required for infection to spread from one person to another:
1. One person must be infected with microorganism.
2. The other person must be susceptible to infection with that microorganism
3. The microorganism must be able to leave the body of the infected person and
enter the body of the susceptible person.
Types of infection
 Colonization – infection present on surface of the body-
Organism propagating at a rate sufficient to maintain its numbers without producing
identifiable evidence of any reaction in host.
 Inapparent –or subclinical infection – organism not only multiplying but also causes a
measurable reaction that is however not clinically detectable
 Symptomatic infection – organism causes clinically detectable reaction.

Development of infection: Onset and Course

  Incubation period –organism present ,no clinical signs,symptoms
 Prodromal period –symptoms ,don’t feel like yourself
 Acute period-fully developed infection.
Clinical Signs and Symptoms
 Local sign
_inflammation
_Purulent exudate if bacteria infection; serous exudate if viral
_Tissue necrosis
_Lymphadenopathy
_Respiratory effects
 Systemic signs:
_Fever,fatigue,headache,nausea
Generallized Stages of infection:
 1. Entry of pathogen
_Portal of Entry
2. Colonization:
_Usually at the site of entry
3. Incubation period
_Asymptomatic period
_Between the initial contant with the microbe and the appearances of the first symptoms.
4. Prodromal symptoms
_Initial symptoms
5. Invasive period:
_Increasing Severity of symptoms
_Fever
_Inflammation and Swelling
_Tissue Damage
_Acme
Modes of Transmission
Modes of spread
Two sources of infection:
 Endogenous or self infection –organism which are the harmless in one site can be
pathogenic when transferred to another site e.g, E.Coli
 Exogenous or cross-infection –organisms transmitted from another source e.g, nurse
doctor and other patient,environment.

Steps to Minimize Risk of Infection:

 Locate, remove reservoir host
 Block portal exit of microbes from reservoir
 Know modes
 Block portals of entry
 Cleaning
 Sterilization
 Disinfectants
 Antiseptics
Hospital acquired infection
 Infection which was neither present nor incubating at the time of admission.
 Includes infection which only becomes apparent after discharge from hospital but
which was acquired during hospitalisation.
  Also called nosocomial infection.
1ST Principle of infection prevention
At least 35-50 % of all nosocomial infection are associated with patient care practices:
 Use and care of urinary catheters
 Use and care of vascular access lines
 Therapy and support of pulmonary function
 Experience with surgical procedures
 Hand hygiene and standard preacautions
Basic steps in prevention of infection

There are possible treatment and prevention to stop the infection cycle. This is through
adequate hygiene, sanitary environment maintenance and health education.
Antimicrobial agents in infection:
 Anti-infective drugs such as antibiotics, antiviral,antifungal and ant tubercular drugs
suppress infection. It can be administered by mouth,topically or intravenously
depending on the infection extent and severity.

 Resident flora
The microorganisms that usually occupy a particular body site are called the resident flora.
Cells of the resident floral outnumber a person own cells 10 to 1. Microorganism that
colonize people for hours to weeks but do not establish themselves permanently are called
transient flora.
Resident flora affect human health:
The normal flora prevent colonization by pathogens by competing for attachment sites or for
essential nutrients. This is thought to be their most important beneficial effect,which has been
demonstrated in the oral cavity,the skin, and the vaginal epithelium.
Protection provided by resident flora:
Rather than causing disease the resident flora often protects the body against disease-causing
organisms. However , under certain conditions,microorganisms that are the part of a person’s
resident flora.
Condition causing diseases:
 The use of antibiotics (ANTIBIOTIC RESISTANCE!!)
 Injury of surgery
 A weakened immune system
1:The use of antibiotics:
When antibiotics used to treat an infection kill a large proportion of certain types of
bacteria of the resident flora,other resident bacteria or fungi can grow unchecked.
For example:
If women take antibiotics for a bladder infection, the antibiotics kill some of the resident
flora,allowing yeast in the vagina to multiply and cause a vaginal yeast infection.

2: Injury of surgery
Injury or something surgery can allow resident flora to enter areas that are not supposed
to have bacteria and cause infection.
For example:
A cut on the skin can allow resident skin flora to cause an infection under the skin.
Surgery on the large intestine to spill into sterile areas in the abdomen and cause very
serious infection.
3: A weakened immune system:
As occurs in people with AIDSor cancer, people taking corticosteroids, and those
receiving cancer(chemotherapy)
Role of the resident flora:
The members of the normal flora play a role both in the maintenance of the health and in
the causation of disease in three major ways
1: They can cause disease,
Especially in immunocompromised and debilitated individuals. Although these organism
are nonpathogens in their usual anatomic locations.
2: The create a protective host defense mechanism:
The nonpathogenic resident bacteria occupy attachments sites on the skin and moucosa
that can interfer with colonization by pathogenic bacteria.
 3:They may serve a nutritional function:
The intestinal bacteria produce several B vitamins and vitamins k. poorly nourished
people who are treated with oral antibiotics can have vitamins deficiencies as a result of
the reduction in the normal flora.

Functions of the Resident Flora:

It should be appreciated that microbes serve a useful purpose in their human hosts. The normal
microbiota maintains a pro-tected envvironment that prevents colonization with potentially
pathogenic organisms. For example: clostridium difficile pro-duces gastrointestinal disease when
the normal intestinal flora have been removed by antibiotics.
The normal microbial flora is more or less constant for each mammalian species and is broadly
divided into resident and transients.
The interction between the microbes and human can result in the following general outcomes: (a)
disease, (b) transient colonization, (c) prolonged colonization
 MECHANISM OF INVASION
The invasion of a host by a pathogen may be aided by the production of bacterial extracellular
substances which act against the host by breaking down primary or secondary defenses of the
body.

Medical microbiologists refer to these substances as invasions.

Invasions are proteins (enzymes) that act locally to damage host cell and or have
the immediate effect of facilitating the growth and spread of pathogens.
The damage to the host as a result of this invasive activity may become part of the pathology of
an infectious disease. Invasions are not clearly distinguished from some extracellular protein
toxins(exotoxins) which also damage the host
Invasion usually act as a short range (in the immediate vicinity of the bacterial growth) and
may not actually kill cells in their range of activity; exotoxins are often cytotoxic and may act at
remote site (removed from the site of bacterial growth)
Also exotoxins typically are more specific and more potent in their activity than invasions
Even so, some classic exotoxins (e.g diphtheria toxin, anthorax toxin) may play some role in
invasion in the early stages of an infection and some invasions (e.g staphylococcal leukocidin)
have a relatively specific cytopathic effect.
Spreading Factors
Spreading Factors describes a family of bacterial enzymes that effect the physical
properties of tissue matrix and intracellular spaces, thereby promoting the spread of the
pathogens

Hyaluronidase:
It is the original spreading Factor. It is produced by Streptococci, staphylococci and clostridia.
The enzyme attacks the interstitial cement (ground substance) of connective tissue by
depolymerizing hyaluronic acid (polysaccharides)
Collagenase:
It is produced by clostridium histolyticum and clostridium perfringens.
It breakdown collagen the frame work of muscles, which facilitates gas gangrene due to these
organisms.
Neuraminidase:
It is produced by intestinal pathogens such as Vibrio cholera and Shigella dysenteries.
It degrades neuromeric acid (also called sialic acid) an intercellular cement of the epithelial
cells of the intestinal mucosa.
Streptokinase & Staphylokinase:
These are produced by Streptococci and staphylococci, respectively.
Kinase enzymes convert inactive plasminogen to plasmin which digests fibrin and prevent
clotting of the blood.
 The relative absence of fibrin in spreading of bacterial lesions allows more rapid
diffusion of the infectious bacteria.

 CLASSIC STAGES OF CLINICAL INFECTION

An infection is the successful colonization of a host by a microorganism.

Infections can lead to disease, which causes signs and symptoms resulting in a deviation
from the normal structure or functioning of the host. Microorganisms that can cause
disease are known as pathogens.

The signs of disease are objective and measurable, and can be directly observed by a
clinician. Vital signs, which are used to measure the body’s basic functions, include body
temperature (normally 37 °C [98.6 °F]), heart rate (normally 60–100 beats per
minute), breathing rate (normally 12–18 breaths per minute), and blood pressure
(normally between 90/60 and 120/80 mm Hg). Changes in any of the body’s vital signs
may be indicative of disease.

The five periods of disease (sometimes referred to as stages or phases) include:

1. Incubation
2. Prodromal
3. Illness
4. Decline
5. Convalescence

INCUBATION PERIOD
The incubation period occurs in an acute disease after the initial entry of the
pathogen into the host (patient).
The incubation period occurs in an acute disease after the initial entry of the pathogen
into the host (patient). It is during this time the pathogen begins multiplying in the host.
However, there are insufficient numbers of pathogen particles (cells or viruses) present
to cause signs and symptoms of disease.
Incubation periods can vary from a day or two in acute disease to months or years in
chronic disease, depending upon the pathogen. Factors involved in determining the length
of the incubation period are diverse, and can include strength of the pathogen, strength
of the host immune defenses, site of infection, type of infection, and the size
infectious dose received. During this incubation period, the patient is unaware that a
disease is beginning to develop.

PRODROMAL PERIOD
The prodromal period occurs after the incubation period. During this phase,
the pathogen continues to multiply and the host begins to experience general signs and
symptoms of illness, which typically result from activation of the immune system, such
as fever, pain, soreness, swelling, or inflammation. Usually, such signs and symptoms
are too general to indicate a particular disease.
 ILLNESS PERIOD
Following the prodromal period is the period of illness, during which the signs and
symptoms of disease are most obvious and severe.

DECLINE PERIOD
The period of illness is followed by the period of decline, during which the
number of pathogen particles begins to decrease, and the signs and symptoms of illness
begin to decline. However, during the decline period, patients may become susceptible to
developing secondary infections because their immune systems have been weakened by
the primary infection.

CONVALESCENCE PERIOD
The final period is known as the period of convalescence. During this stage, the
patient generally returns to normal functions, although some diseases may inflict
permanent damage that the body cannot fully repair
 STERILIZATION
Sterilization refers to any process that removes, kills, or deactivates all forms of
life (in particular referring to microorganisms such as fungi, bacteria, viruses, spores,
unicellular eukaryotic organisms such as Plasmodium, etc.) and other biological
agents like prions present in a specific surface, object or fluid, for example food or
biological culture media.
APPLICATION

FOOD:
One process by which food is sterilized is heat treatment. Heat treatment ceases
bacterial and enzyme activity which then leads to decreasing the chances of low quality
foods while maintaining the life of non-perishable foods.

MEDICINE AND SURGERY:

Surgical instruments and medications that enter an already aseptic part
of the body (such as the bloodstream, or penetrating the skin) must be sterile. Examples
of such instruments include scalpels, hypodermic needles, and artificial pacemakers. This
is also essential in the manufacture of parenteral pharmaceuticals.
 STEAMING:
Steam sterilization is the most widely used and the most dependable. Steam
sterilization is nontoxic, inexpensive, rapidly microbicide, sporicidal, and rapidly heats
and penetrates fabrics.

TYPES OF STERILIZATION
Following are the types of sterilization:
1. Chemical sterilization
2.Radiation sterilization
3. Non-ionizing radiation sterilization

CHEMICAL STERILIZATION:
Chemical sterilization is the process of using low temperature chemicals to
kill, eliminate, and remove all germs, viruses, and bacteria. This can be in the form of gas
or liquid chemicals.
 Radiation sterilization:
Sterilization can be achieved using electromagnetic radiation, such as Ultraviolet light,
X-rays and gamma rays, or irradiation by subatomic particles such as by electron beams.[55]
Electromagnetic or particulate radiation can be energetic enough to ionize atoms or molecules
(ionizing radiation), or less energetic (non-ionizing radiation.

Non-ionizing radiation sterilization:

Ultraviolet light irradiation (UV, from a germicidal lamp) is useful for sterilization of surfaces
and some transparent objects. Many objects that are transparent to visible light absorb UV. UV
irradiation is routinely used to sterilize the interiors of biological safety cabinets.
  DISINFECTION
Disinfection describes a process that eliminates many or all pathogenic
microorganisms, except bacterial spores, on inanimate objects.
Low-level disinfectants can kill most vegetative bacteria, some fungi, and some viruses
in a practical period of time (≤10 minutes).

Importance of Disinfection
To minimize number of organisms in the population worldwide. The method of disinfection is
used internationally for the safety of humans, to decrease the scale of transmission of diseases.
A large emphasis of sterilization and disinfection has been placed in the food industry, water
sanitization and medical care and hospitals. As these have found to be largest affected
organization’s with microorganisms and modes of transmission amongst the population.
Different disinfectants are used in different industries, which target the specific flora.

Disinfection Techniques
Consistency
• Liquid (Alcohols, Phenols)
• Gaseous (Formaldehyde vapor, Ethylene oxide)
Spectrum of activity
• High level
• Intermediate level
• Low level
Mechanism of action
Action on membrane (Alcohol, detergent)
• Denaturation of cellular proteins ( Alcohol, Phenol)
• Oxidation of essential sulfhydryl groups of enzymes (H2O2, Halogens)
• Alkylation of amino-, carboxyl- and hydroxyl group (Ethylene Oxide, Formaldehyde)
• Damage to nucleic acids (Ethylene Oxide, Formaldehyde
Uses of Disinfection
Aldehydes: surface disinfection, fumigation of rooms, chambers and operating theatres.
Alcohol: 70% aqueous alcohol is more effective at microbial killing. 70% Ethyl alcohol is used
as antiseptic on skin.
Phenol: first used by Lister to prevent infection in surgical wounds. In high concentrations used
as a disinfectant and in low concentrations as an antiseptic.
Halogens: Iodine (antiseptic), Chlorine (bleach)