TA SERIES

MICROBIOLOGY
SLIDES BOOK 2023
BSN 1ST SEMESTER
KMU

COMPILED BY:
TUFAIL
BSN 2022-2026
GINAHS D.I.KHAN
CONTENTS
S.NO CHAPTER PAGE NO
1. Introduction to Microbiology 01
2. Control of Microorganism 12
3. Body Defence System 30
4. Medical and Surgical Asepsis 36
5. Concept of Isolation 44
6. Human and Microbial Intrection 51
7. Microbiology in Everyday life 71
8. Microscope Use (LAB WORK) 76
9. Staining of Bacteria (LAB WORK) 78
10. Safe Use of Lab (LAB WORK) 81
11. Culture Media Slides (LAB WORK) 85

FOR FEEDBACK AND SUGGESTION : sta1372002@gmail.com

 Unit 01 Introduction to Microbiology
Microbiology
 The term microbiology derives its name from three Greek words micros [small] bios [life] and logos
[study].
 Microbiology is the science that deals with the study of microorganisms.
 Microbiology is the study of those organisms and agents that are too small to be seen clearly by the
unaided eye.
 To be more simple, microbiology is the study of microorganisms which are the living organisms of
microscopic size.
Microorganism
֍ Microorganisms are the living organisms that are less than 1 millimeter in diameter which cannot be
seen by our naked eye.
֍ Microorganisms can be viewed through microscopes and they can exist as single cells or clusters.
֍ Microorganisms include the cellular organisms like bacteria, archaea, fungi, algae and protozoa.
֍ Viruses and bacteriophage are also included as one of the microorganism but they are acellular.
Sub disciplines of microbiology
 Bacteriology The study of bacteria  Virology The study of viruses
 Protozoology The study of protozoa  Immunology The study of immunity
 Mycology The study of fungi  Phycology The study of Algae
Terminology
o Pathogen: An organism which causes disease is called pathogen.
o Parasite: An organism that lives in or on another species or creature and obtains food and shelter
without benefiting but rather harming the host.
o Saprophyte: An organism that lives on or derives its food from dead organic matter. e.g. Fungi
o Normal flora: The microbes that live in or on another creature and benefiting each other in normal
conditions.
o Vector: An organism usually an arthropod such as insect or tick that transfers a pathogen to a person.
1) Mechanical vector and 2) Biological Vector.
o Infection: The entry of microorganism specially pathogen in the body is known as infection.
o Incubation period: The period of proliferation of microorganisms to show sign and symptoms after their
entry in a body is known as incubation period.
o Prodromal period: It is the interval between the onset of symptoms of an infectious disease and the
appearance of characteristic manifestations.
o Prognosis: The prediction of the course of a disease.Conti…..
o Epidemiology: The study of occurrence of disease, how, when and where it occurs and how it is
transmitted.
o Epidemic disease: An unusual sudden onset of a disease which breaks out in a region of a country. eg.
Cholera, or any other disease broken out .
o Endemic disease: A disease which is usually found in an area in a country. eg, typhoid, etc.
o Pandemic disease: An epidemic disease that occurs worldwide. e.g. Corona Virus.
o Sporadic disease: A disease which is found here and there at intervals. eg. T.B, etc.
o Isolation: separation of infected persons for a period of communicability of the disease.
o Quarantine: limitation of the movement of apparently well person or animal who has been exposed to
the infectious disease for a duration of the maximum incubation period of the disease.
o Inflammation: The response of body to infection or injury which is characterized by swelling, heat,
redness and pain.
o Pathogenicity: The ability of an organism to cause disease.
o Virulence: The degree or intensity level of a pathogen.
o Toxin: A poisonous substance produced by a living organism.
o Toxicant: A poisonous substance prepared by man.
o Antigen: Any sustance usually made up of protein that stimulates the immune response.
o Antibody:Aprotein that is formed as a result of the immune response to an antigen.
1
o Sterilization: The process by which all forms of life is killed.
o Bactericide: A substance or agent that kills bacteria.
o Disinfectant: An agent that kills microorganisms by applying to inanimate (non living) objects.
o Antiseptic: An agent that kills microorganisms by applying to living things.
o Vaccine: Attenuated or killed microbes or inactivated toxins used to induce immunity.
o Vaccination: The process of inducing immunity by administrating a vaccine.
o Fomites: Inanimate object or material used by an infected patient, which may transfer the infection to
another person by coming in contact. For example, Lenin, clothes and other utensils.
o Eukaryotic Cell: A complete cell having a well defined nucleus and other membranous structures. eg.
Multicellular(like animal and higher plant cells) and unicellular (like protozoans).
o Prokaryotic Cell: An incomplete cell having no well defined nucleus and other membranous structures.
They are always unicellular. eg. Bacterial cell .
o Bacteria: They are prokaryotic microorganisms which can’t be seen with naked eyes.
o Virus: A minute microorganism which shows the characteristics of both living and non living things. It is
an obligatory parasite.
o Protozoa: Microscopic single-celled eukaryotic microbes. Some are
pathogenic and some are not.
Importance of Microbiology in Nursing
For Nursing student ,microbiology is a vital subject. Nurses are involved in managing all aspects of patient’s
health and infection control in the hospitals. Nurse must know microbiology to take care of patient and to
protect oneself from pathogenic microorganisms. Students who major in medical microbiology need to
know more about the bacteria that cause human illnesses, the disease they cause ,as well as how to
diagnose ,prevent and treat them.
Role of nurses in different units
1.To prevent spread of infection 2.To maintain sterile field
3.To collect specimen 4.To implement immunization schedule in hospital
5.Implementation of microbiology in using:Role of nurses in different units
 Burns unit  Operation theatres
 Obstetric units  Intensive care nursery
7. To dispose of biomedical waste
To prevent spread of infection
Infections propagate in different ways, and nurses should be aware of this. As well as contact (touch),air
(airborne),droplets (sneezing, coughing),contaminated food and drink (foodborne),sexual contact, and
infected blood transfusion,etc.
֍ Nurses can utilize this knowledge to identify specialized infection control
measures.
By understanding how disease-causing
organism enter ,exit and spread from
one individual to another, nurses can
take special measures to prevent
infections within the hospital and
community.
 Example of such diseases include
cholera, common cold,chiken pox, and TB.
 When a person suffering from common cold sneezes, fine droplets of moisture, carrying thousands of
viruses are spread in the air. The virus may enter the body of healthy person while breathing.
To maintain sterile field
֍ In hospital ,sterile fields must be established and maintained using microbiology techniques ,which a
nurse needs to be familiar with. Nurses must understand how to create and maintain sterile fields in
operating rooms, for example ,in order to prevent infection following surgery.

2
֍ Remember to wash your hand before and after surgeries . Nurses benefit from hand washing by
minimizing the number of microorganism on their hands, thereby reducing their risk of hospital acquired
infection.
֍ The nurses can also use microbiology knowledge to use sterile equipment ,which is necessary for
invasive procedures. Sterilization process need to be understood by nurses.
֍ The nurses can also use microbiology knowledge to use sterile equipment ,which is necessary for
invasive procedures.
To collect specimens
֍ Nursing practitioners must understood the importance of properly collecting specimen for
bacteriological testing in order to achieve reliable results.
֍ He/she must be familiar with a variety of infectious diseases and their mood of infection.
֍ A nurse must know how importance it to collect specimens correctly for bacteriological analysis in order
to obtain reliable results. In order to identify the organism responsible for an infection ,it is necessary to
collect a clinical specimen from the appropriate site of infection.
֍ A nurse must know how importance it to collect specimens correctly for
bacteriological analysis in order to obtain reliable results.
To implement immunization schedule in hospital
֍ In addition to immunization patients against diseases such as diphtheria and MMR,nurses also play an
important role in preventing other diseases.
֍ A nurse should be knowledgeable about vaccine schedules and how vaccines are transported from
production to administration via a cold chain.
֍ Microbiology is a subset of immunology that can assist nurses in this endeavor.
֍ The nurse who know immunology is well prepared for vaccines and is able to use the cold chain
effectively for vaccination and vaccine protection.
Implementation of microbiology in nursing: Role of nurses in different units
֍ Some of the areas discussed in health care setting relate to microbiology role as a nursing specialty.
֍ Patients with contagious or infectious diseases are hospitalized in contagious or infectious wards.
֍ For example the rabies wards or the gastroenteritis wards ,and the nurse take great care of them by
taking precautions to prevent the infection from spreading others.
֍ The nurse employ aseptic procedure while caring for patients.
֍ Burns’ unit: Initially a burn wound is sterile but within a short time the microorganisms starts colonizing
in it. A nurse takes special care to prevent the wound from becoming infected when caring for such type of
patient. She follows not only aseptic techniques but also uses sterile equipment while looking after such
patients.
֍ Operation theatres: Before operation, it is duty of a nurse to ensure that the atmosphere of the
operation theatre (OT) is free of microorganisms. It is done by the process of fumigation of the operation
theatre from time to time. Nurse should assure that all procedures on a patient are done under sterile
conditions. Swabs are cultured from a variety of locations throughout the operation theatre to determine a
theatre,s sterility.it is imperative that procedure on a patient is conducted under sterile conditions
֍ Obstetric units: The wound infection of birth canal after child birth or abortion is a major cause of
maternal mortality rate. The reason is that the infection spreads and causes peritonitis or phlebitis. The
nurse can play a role during antenatal care, help during delivery or after giving birth for 6 weeks period
called puerperium.
֍ Intensive care nursery: In intensive care nurseries especially experienced staff is employed who has
good knowledge of incubators, gastrostomy feeding, exchange transfusion, respirator and umbilical
catheters which are necessary for infants. Nurses must keep a check to maintain sterile atmosphere in
these intensive care nurseries.
To dispose of biomedical waste
֍ There must be an appropriate way to dispose of biomedical waste, which is based upon microbiology
principles.
֍ A nurse should be familiar with the process used in hospital to create and maintain sterile fields based
on microbiological expertise.
3
֍ Biomedical waste must be disposed of. If biomedical waste is not properly disposed of, it may result in
the outbreak of a terrible disease .
֍ Proper biomedical waste dispose based on microbiology principle is required.
֍ Biomedical waste must be disposed of. If biomedical waste is not properly disposed of, it may result in
the outbreak of a terrible disease .
History Altering Microbes
 Although not always recognized at the time, Microbes have dramatically altered human history.
 In history ,1347 plague black death (Bubonic plague) struck Europe, only after four year later the plague
had killed 1/3 of the population (about 25 million people).
 Over the next 80 year the disease struck again ,eventually wiping out 75% of the European population.
 Bubonic plague-Yersinia pestis (Bacterium).  Black Death(1347-1351) .
 Vector borne septicemia.  Malaria –plasmodium falciparum (protozoan)
 Mosquito –borne infection disease  Thought to have killed Alexander the Great (323 BC)
 Still an important pathogen  Infects-400,000,000 annually
 Kills -2,000,000 annually  That is over 5,000 every day.
 Potato Blight- phytophthora Infestans (Fungus)  Irish potato famine(1845-1849)
 Potato crop were wiped out  1,000,000 starved to dead
 2,000,000 emigrated  Many came to the united states in the mid-1800s
 Still a chronic problem for potato formers.
History of Microbiology
 Major contribution to the development of microbiology was the invention of the microscope by Anton
von Leuwenhoek and the implementation of the scientific method.
 Concept of microbiology started from 1600s.  Institution Contributions
 Pasteur Institute  CDC-Center of disease control
 NIH-National Institute of health  WHO-word health organization
Antony Van Leeuwenhoek(1632-1723)
o (Dutch lens maker) Antoine Van Leeuwenhoek o Simple microscope 50*300X magnification
o First observation of live microorganisms by microscope by his own design .
o Made and reported many detailed observations.
o Named the observed tiny object as ―animalcules, in 1676.
o Laid the foundation of bacteriology and protozoology.
o Book – (The secrete of nature) o He also discovered sperm .
Contribution of Leeuwenhoek
o He constructed the first microscope. o The first person to observe microorganism.
o He first accurately described the different shape of bacteria.
o It is clear from his description that he saw both bacteria and protozoa.
o Microorganism were first seen by Leeuwenhoek (1673) and he found many microorganism in material
such as water, saliva , and the intestinal contents, of healthy subject, and he recognized them as a living
creatures.
o He is regarded as Father of „Bacteriology‟ and „Protozoology‟, because of his contribution to the field of
bacteria and protozoa.
Francesco Redi
o Francesco Redi (18 February 1626 – 1 March 1697) was an Italian physician , naturalist, biologist and
poet.
o He is referred to as the "founder of experimental biology, and as the "father of modern parasitology".
o From earliest time ,people had believed in spontaneous generation (Abiogenesis) even Aristotle
supported this idea.
o He was the first person to challenge the theory of spontaneous generation by demonstrating that
maggots come from eggs of Flies.
Francesco Redi Experiments
o Redi placed meat in three container ,one was uncovered, a second was covered with paper, and the third
was a fine gauze that would exclude flies.
4
o Flies laid their eggs on the uncovered
meat and maggots developed.
o The other two pieces of meat did not
produce maggots spontaneously.
o However ,flies were attracted to the
gauze-covered container and laid their
eggs on the gauze ,these eggs produced
maggots, hence spontaneous generation
was disapproved. Francesco Redi experiment
Louis Pasteur(1822-1895)
o He was a French Biochemist, (1822-1895).
o He is regarded as „Father of Microbiology and Immunology‟.
o He proposed the „Theory of Germ Disease‟, where diseases of plants, viruses, animals and human beings
are caused by pathogenic microbes.
o He disproved the theory of abiogenesis by conducting „Swan neck flask experiment‟.
o Proved that all form of life even microbes arose only from their likes (disapproved spontaneous
generation).
Contribution of Louis Pasteur
o He coined the term ,microbiology, and also discovered the presence of bacteria in the air and classified
the bacteria into aerobic and anaerobic forms.
o Established that fermentation (chemical break down of Substance by microorganism )was the result of
microbial activity.
o He developed technique to prevent souring of milk and spoilage of wine. His technique is now called
Pasteurization technique.
o Process of attenuation discovered and production of attenuated vaccine.
o He first isolated bacteria causing cholera (Vibrio cholera). o Various antiseptic technique.
Develop sterilization Technique Hot Pasteur's Experiment Swan-necked flask experiment
air oven Autoclave Steam Sterilizer

Contribution of Robert Koch

o He was a German microbiologist
(1843-1910).
o His contribution to the field of
microbiology and medical science
is the most valuable one.
o He developed for the first time
staining technique to stain the
bacteria with acidic or basic stain.
o He developed for the first time
culture technique to culture the
bacteria in the laboratory.
o He discovered bacteria caused
tuberculosis of man. Koch Postulates
5
Contribution of Robert Koch
o He isolated and identified different kinds of bacteria from various sample.
o He was awarded Nobel Prize of medicine in 1905, formulating principles regarding diseases. These are
now called „Koch Postulates‟.
o He proved theory of germ diseases of Louis Pasteur by conducting investigative experiment.
o His criteria for proving the causal relationship between a microorganism and a specific disease.
Prokaryotic Cells
o Prokaryote is a Greek word, pro - before and karyon - nut or kernel.
o Prokaryotes are the organism with a primordial nucleus.
o They have a much simpler morphology than eukaryotic cells and lack a
true membrane bound nucleus and cell organelles like mitochondria, Golgi
bodies, endoplasmic reticulum, etc.
o All bacteria and archaea are prokaryotic.
Eukaryotic Cells
o Eukaryote is a Greek word, Eu - true and karyon means - nucleus.
o Eukaryotes posses a membrane enclosed nucleus and cell organelles.
o Eukaryote cells are the cell that are complex in structure and function as
they have a membrane bound well defined nucleus and other membrane
bound organelles.
o Algae, fungi, protozoa, higher plants and animals are eukaryotic.
 Eukaryotes are organisms whose cells have a nucleus and other organelles enclosed by a plasma
membrane .
 Organelles are internal structures responsible for a variety of function ,such as energy production and
protein synthesis.
 Eukaryotic cells are large (around 10-100 um) and complex.
 While most eukaryotes are multicellular organisms, there are some single-cell eukaryotes.
Difference b/w Prokaryotic & Eukaryotic cell

Virus
Def : A virus is a submicroscopic infectious agent that replicate only inside the living cells of the organism,
is a small parasite that can not reproduce by itself. Viruses infect all type of life forms ,from animals and
plant to microorganism ,including bacteria and archaea.
Size:  A small virus has a diameter of about 20nm e.g. Parvovirus
 A large virus have a diameter of upto 400nm e.g. poxvirus
Medical Importance Of Viruses
Viral disease range from minor
aliments such as the common
cold to terrifying disease such as
rabies or AIDS.

6
They may be sporadic like mumps
֍ Endemic like infectious hepatitis ֍ Epidemic like dengue fever ֍ Pandemic like influenza,COVID-19.
Viruses can cause cancer in animal and birds, as well as in human.
Virus structure & its basic properties

Viruses that commonly Cause Diseases

Portal of En Virus Disease
Respiratory Tract Influenza virus Influenza
Rhinovirus Common Cold
Epstein- Barr virus Bronchiolitis
Cytomegalovirus Mononucleosis syndrome
Measles Virus Measles
Mumps Virus Mumps
Rubella virus Rubella
Hantavirus Pneumonia
Adenovirus Pneumonia
Gastrointestinal tract Hep A Virus Hepatitis
Polio virus Poliomyelitis
Rota virus Diarrhea
Skin Rabies Virus Rabies
Dengue Virus Dengue
Yellow Fever virus Yellow Fever
Blood Hep B Virus Hepatitis B
Hep C virus Hepatitis C
Hep D virus Hepatitis D
HIV AIDS
Human T Cell Lymphotropic Virus Leukemia
Genital Tract Hep B Virus Hepatitis B
HIV AIDS
Herpes simplex Virus Herpes genitals and neonatal
herpes
Trans placental Cytomegalovirus Congenital Abnormalities
Rubella Congenital Abnormalities
Parvovirus Hydrops Fetalis

DNA and RNA Viruses

DNA Viruses RNA Viruses
Small pox Virus Polio Virus
Herpes Virus Mumps Virus
Hepatitis B virus Measles Virus
Pox virus HIV virus
7
DNA Viruses RNA Viruses
Adeno virus Influenza Virus, Hepatitis C Virus
Rubella Virus, Dengue Virus
CHARACTERISTICS OF BACTERIA
 Antoni van Leeuwenhoek Discovered bacteria in 1676.
 Bacteria are prokaryotic unicellular organisms
 Bacteria are found by almost everywhere on Earth
 In other hand bacteria is destructive and cause disease like pneumonia,TB.
 These organism can live in soil the ocean inside the human gut almost everywhere.
 Truly speaking, all of the bacteria are potentially harmful. Any how, about 97 % bacteria are beneficial
and 3 % harmful. Beneficial bacteria include normal flora and other industrial and environmental bacteria .
 Division (reproduction) by Binary fission  Rigid cell wall containing peptidoglycan
 No mitochondria and nuclear membrane  DNA and RNA both are present
 They are 1-10 um in length.  Helping in our digestion.
 Lack a defined Nucleus
Bacterial Classification(Based on Morphology) Shapes of Bacteria

Shapes of bacteria
 Typical bacteria measure 2-8 m in length and 0.2-2 m in width.
 Form associations such as chains, clusters, and tetrads.

Bacterial Structure
Classification of Bacteria

8
A Typical Bacterial Features
 Flagella (Singular: flagellum) are thread like structures for locomotion in most motile bacteria.
 Pili (singular: pilus) Glycoprotein appendages on bacterial cell Shorter than flagella used for transfer of
genetic material from one to another (sex pili).
 Fimbriae (singular: Fimbria) Glycoprotein appendages on bacterial cell Shorter than pili used for
attachment to contact surfaces.
 Capsule - Made of polysaccharide which protects bacteria from phagocytes.the capsule is found most
commonly among gram-negative bacteria: Escherichia coli .
 Cell wall: Made up of peptidoglycan. – Responsible for the rigidity of bacterial cell. (Refer to
peptidoglycan chemical structure on white board)
 Cell Membrane: – Inner to cell wall, there is a delicate cytoplasmic membrane which surrounds the
cytoplasm.
 Outer Membrane:
Exterior to cell wall found in Gram Negative bacteria.
 Ribosome made up of RNA and protein. It is a factory of protein synthesis.
 Mesosome An invagination of cell membrane which helps in cell division.
 Nucleoid (Genetic material) Unlike virus, bacteria have both DNA and RNA. It contains genetic
characteristics.
 Periplasm is the space between cytoplasmic membrane and outer membrane of Grm-ve bacteria which
contains hydrolytic and Beta lactamase enzyme to degrade substances like penicillin.
 Plasmid A fragment of extra chromosomal DNA segment which contains different genes for resistance
to heat and chemicals.
Different Nutritional Types of Bacteria
• Nutrition is substances used in biosynthesis and energy production and therefore are required for all living
things.
• Bacteria, like all living cells, require energy and nutrients to build protein and structural membranes and
drive biochemical process.
• Bacteria require sources of carbon ,nitrogen,phosphorous,iron and large amount of other molecule.
• Carbon, nitrogen, and water are used in highest quantities.
• Some types of bacteria must consume pre-formed organic molecule to obtain energy, while other bacteria
can their own energy from organic sources.
Microbial Nutrition

 The first six(C,O,H,N,S, and P): component of carbohydrate, lipid, and proteins.
 K : Enzyme –protein synthesis.  Ca: Heat resistance od bacterial endospores.
 Mg: Cofactor , stabilize the cell membrane and ribosomes.
 Fe: Part of cytochrome and cofactor for enzyme and ECP.
Extra Point :

9
Microbial Nutrition

Based on Mode of nutrition and Growth

10
Based on Mode of Nutrition and Growth

Some Medically Important Bacteria

 Gram positive Bacteria.
 Streptococcus—causes pneumonia, pharyngitis, cellulitis
 Clostridium (spore forming rods)— Tetanus, botulism
 Staphylococcus—abscess of skin and other organs,
 Bacillus (spore forming rods)—causes Anthrax  Actinomyces— Actinomycosis
 Corynebacterium—diphtheria  Listeria—MeningitisConti…..
 Gram Negative Bacteria.
 Escherichia-- Urinary tract infection, diarrhea  Neisseria---- Gonorrhea, Meningitis
 Bordetella---- Whooping cough  Pseudomonas Pneumonia, UTI
 Salmonella--- Typhoid fever  Shigella ------ Enterocolitis
 Haemophilus- Meningitis  Vibrio--------- Cholera
 Yersinia------ Plague
 Acid Fast
 Mycobacterium--- Tuberculosis, Leprasy
Extra Point :

11
 TA SLIDES BOOK SERIES
UNIT 02 Control of Microorganisms
INTRODUCTION
 Control of microbial growth means to inhibit or prevent growth of microorganisms. Control of microbial
growth usually involves the use of physical or chemical agents which either kill or prevent the growth of
microorganisms.
 Physical agents include such methods of control as high or low temperature, desiccation, radiation, and
filtration.
 Control by chemical agents refers to the use of disinfectants, antiseptics and antibiotics.
REASONS FOR CONTROLLING MICROORGANISMS
Reasons for Controlling Microorganisms
 Prevent contamination.  Prevent transmission of pathogen
 To prevent decomposition & spoilage of products
 To prevent contamination in aseptic areas, processes like production of pharmaceuticals by
fermentation
 To maintain aseptic condition in operation theaters, filling area of non sterile pharmaceuticals.
ACTION OF MICROBIAL AGENTS
 Primary targets of microbial control are the microorganisms that can cause infection or spoilage of food
that are constantly present in the external environment.
 Modes of action fall into two basic categories.
֍ Alteration of cell walls or cytoplasmic membranes
֎ Interference with protein and nucleic acid structure
ALTERATION OF CELL WALL AND MEMBRANES
 Cell wall maintains integrity of cell
֎ When disrupted, cannot prevent cell from bursting due to osmotic effects
 Cytoplasmic membrane contains cytoplasm and controls passage of chemicals into and out of cell
֎ When damaged, cellular contents leak out
 Viral envelope responsible for attachment of
virus to target cell
֎ Damage to envelope interrupts viral replication.
GENERAL TERMS
 Sterilization:
Sterilization is the process of destroying all living
organisms and viruses. A
sterile object is one free of all life forms, including
bacterial endospores, as well as viruses.
 Disinfection:
Disinfection is the elimination of microorganisms, but not necessarily
endospores, from inanimate (non-living) objects or surfaces.
 Decontamination:
De-contamination is the treatment of an object or inanimate surface to make it safe to handle.
 Disinfectant:
A disinfectant is an agent used to disinfect
inanimate objects but generally is toxic to
use on human tissues.
 Antiseptic:
An antiseptic is an agent that kills or inhibits
growth of microbes but is safe to use on
human tissue.
 Sanitizer:
A sanitizer is an agent that reduces microbial
numbers to a safe level.
12
 TA SLIDES BOOK SERIES
 Sepsis:
The presence of Bacteria Contamination.
 Asepsis:
The absence of Bacteria Contamination.
 Aseptic Technique:
Used to prevent microbial contamination of any clinical procedure and products of pharmaceutical and
food industry to keep standards.
 Antibiotic:
Antibiotic is a substance derived from microorganisms or produced synthetically to kill or inhibit the
growth of other microorganisms i.e. bacteria.
 Chemotherapeutic synthetic drugs:
Synthetic chemicals that can be used therapeutically.
 Cidal:
An agent that is cidal in action; will kill microorganisms .
 Static:
An agent that is static in action; will inhibit the growth of microorganisms.
 Bacteriostatic Agent: An agent that inhibits the growth of bacteria e.g. tetracycline, macrolides,
clindamycin etc.
 Bactericidal agent: An agent that kills bacteria e.g.
Aminoglycoside, betalactum, Fluorquinolones etc.
 Germicide: An agent that kills certain
microorganism/ germs
 Virucide: An agent that inactivates/ kill viruses.
 Fungicide: An agent that kills fungi.
 Sporicide: An agent that kills bacterial endospores
and fungal spores e.g.
Sodium hypochlorite (NaClO).
METHODS OF MICROBIAL CONTROL PHYSICAL METHODS

Physical methods:
The methods which are used to destroy or remove microorganisms by means of physical things are known
as physical methods.
Examples:
 Temperature  Radiation  Refrigeration  Desiccation
13
 TA SLIDES BOOK SERIES
1: Temperature:
 Microorganisms have a minimum, an optimum, and a maximum temperature
for growth.
 Temperatures below the minimum usually have a static action on
microorganisms. They inhibit microbial growth by slowing down metabolism but
do not necessarily kill the organism.
 Temperatures above the maximum usually have a cidal action, since they
denature microbial enzymes and other proteins. Temperature is a very common
and effective way of controlling microorganisms.
A. Thermal death point (TDP)
Lowest temperature at which all of the microbes in a liquid suspension will be killed in ten minutes.
B. Thermal death time (TDT)
The time required to kill all bacteria in a liquid culture at a given temperature.
C. Decimal reduction time (DRT)
The time required to kill 90% of the bacteria in a liquid culture at a given
temperature.
i. High temperature:
 Vegetative microorganisms can generally be killed at temperatures from 50°C to 70°C with moist heat.
 Bacterial endospores, however, are very resistant to heat and extended exposure to much higher
temperature is necessary for their destruction.
 High temperature may be applied as either moist heat or dry heat.
B. Moist heat:
Moist heat is generally more effective than dry heat for killing microorganisms because of its ability to
penetrate microbial cells. Moist heat kills microorganisms by denaturing their proteins (causes proteins
and enzymes to lose their shape).
It also may melt lipids in cytoplasmic membranes.
 Boiling water  Autoclaving
Boiling water:
 Boiling is the process of turning a liquid into a vapor and for a certain atmospheric pressure, the boiling
happens at a specific temperature.
 Boiling to 100°C or more. It kill vegetative form of bacterial pathogen, almost all viruses, and fungi.
 Endospores and some viruses are not destroyed quickly.
 Hepatitis virus: Can survive up to 30 minutes of boiling. However brief boiling will kill most pathogens.
Autoclaving:
 Autoclaving employs steam under pressure.
 Reliable sterilization with moist heat requires temperature above that of
boiling water.
 Example of moist heat is autoclave, a chamber which is filled with hot
steam under pressure .this is preferred method of sterilization for non-heat
sensitive materials in which temperature of steam reaches 121°C at 15 psi
pressure.(pound per square inch).
 Autoclaving is cidal for both vegetative organisms and endospores, and is the most common method of
sterilization for materials not damaged by heat.
b. Dry Heat:
Dry heat kills microorganisms through a process of protein oxidation rather than protein coagulation.
Examples of dry heat include:
 Direct Flaming  Hot air Sterilization  Incineration  Filtration
 Direct Flaming:
Used to sterilize inoculating loops and needles. Heat metal until it has a red glow.

14
 TA SLIDES BOOK SERIES
 Filtration:
Removal of microbes by passage of a liquid or air through a screen like material with
small pores .used to sterilize heat sensitive material like vaccine, , antibiotics, and
some culture media. Used in operating rooms and burn units to remove bacteria
from air.
Hot air Sterilization:
Microbiological ovens employ very high dry temperatures: 171°C for 1 hour; 160°C
for 2 hours or longer; or 121°C for 16 hours or longer depending on the volume.
They are generally used only for sterilizing glassware, metal instruments, and other
inert materials like oils and powders that are not damaged by excessive temperature.
Incineration:
 The process of burning waste material at temperature high to destroy
contaminants.
 Three types of waste to which incineration is applied extensively are
municipal solid waste, hazardous waste, medical waste.
 Incinerators are used to destroy disposable materials by burning.
 Incineration is a method of treating waste which involves the combustion
of the organic substances found in waste materials.
c. Pasteurization:
 Pasteurization is the mild heating of milk and other materials to kill particular
spoilage organisms or pathogens. It does not, however, kill all organisms. Milk
is usually pasteurized by heating to 71.6°C for at least 15 seconds in the flash method or 62.9°C for 30
minutes in the holding method.
 This is sufficient to kill the vegetative cells of the milk-borne pathogens (e.g. mycobacterium, salmonella,
streptococcus),but not to sterilize the milk.
 Classic method of Pasteurization:
Milk is heated to 62°C for 30 minutes.
 High Temperature short time pasteurization:
Milk is exposed to 72°C for 15 seconds.
 Ultra high temperature pasteurization:
Milk is treated at 140°C for 3 seconds and then cooled very quickly in
a vacuum chamber.
Advantages: Milk can be stored at room temperature for several months.
ii. Low Temperature:
Low temperature inhibits microbial growth by slowing down microbial metabolism. Examples include:
refrigeration and freezing.
 Refrigeration at 0°C to 7°C slows the growth of microorganisms and keeps food fresh for a few days.
Bacteriostatic effect.
 Freezing items are protected by stopping the growth of microbes at temperature below 0°C , and keeps
food fresh for several months.
2. Desiccation:
It is the process of removal of water. In the absence of water, microbes cannot
grow or reproduce, but some may remain viable for years. After water
becomes available, they start growing again.
3. Radiation:
Two 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 .
 Used to sterilize pharmaceuticals and disposable medical supplies. Food industry is interested in using
ionizing radiation.
15
 TA SLIDES BOOK SERIES
Disadvantages:
 Penetrates human tissues. May cause genetic mutations in humans.
2. Ultraviolet light (Nonionizing Radiation):
 Wavelength is longer than 1 nanometer Damages DNA by producing thymine dimers.
 The greatest antimicrobial activity of UV light occurs at 250 to 260 nm, which is the wavelength region of
maximum absorption by the purine and pyrimidine bases of DNA.
 DNA replication is inhibited and the organism can not grow.
 Used to disinfect operating rooms, nurseries.
Disadvantages: Damages skin and eyes.
CHEMICAL CONTROL OF MICROORGANISMS
Chemical methods of destruction:
 Very few chemical agents actually achieve sterility, but they do reduce the
microbial population to safe levels, destroying pathogen.
 Chemical agents act primarily by one of the three mechanism.
disruption of cell membrane modification of protein modification of DNA
 The following table illustrates some of the commonly used chemical agents.
CHEMICAL METHODS
1: Phenol (carbolic acid):
 was first used by Lister(1860) as a disinfectant.
 Destroys plasma membrane and denatures proteins.
 Effective against gram-positive staphylococci and streptococci.
 Rarely used today because it is a skin irritant and has strong odor.
Advantages: Stable, persist for long times after applied, and remain active in the presence of organic
compounds.
 Disadvantages :Excessive use in infants may cause neurological damage.
2: Halogens:
Effective alone or in compounds.
A. Iodine:
 Tincture of iodine (2% solution of iodine and potassium iodide in ethanol) denatures proteins.
 It Stains skin and clothes.  Used as skin antiseptic in surgery
 Not effective against bacterial endospores
B. Chlorine:
 Used to disinfect drinking water, pools, and sewage.
 Chlorine is easily inactivated by organic materials.
 Sodium hypochlorite is active ingredient of bleach
 When mixed in water, forms hypochlorous acid:
 Cl2 + H2O ------> H+ + Cl- + HOCl
3: Alcohols:
 Kill most bacteria, fungi, but not endospores.
 Act by denaturing proteins and disrupting cell membranes.
 Used to mechanically wipe microbes off skin before injections or blood drawing.
 Not good for open wounds, because cause proteins to coagulate.
 They are bactericidal, and fungicidal but not sporicidal.
4: Heavy Metals:
 Heavy metals and their compounds are microbicidal.
 Include copper, selenium, mercury, silver, and zinc.
 The property of heavy metal to exert biocidal effect is called Oligodynamic action.
A. Silver:
 1% silver nitrate used to protect infants against gonorrheal eye infections until recently.
B. Copper:
 Copper sulfate is used to kill algae in pools and fish tanks.
16
 TA SLIDES BOOK SERIES
C. Selenium
 Kills fungi and their spores. Used for fungal infections. Also used in dandruff shampoos.
D. Zinc
 Zinc chloride is used in mouthwashes.  Zinc oxide is used as antifungal agent in paints.
5: Oxidizing Agents:
A. Ozone:

It is Used to disinfect water.

Helps neutralize unpleasant tastes and odors.

More effective killing agent than chlorine, but
less stable and more
expensive.
B. Hydrogen Peroxide:
 Used as an antiseptic.
 Not good for open wounds because quickly broken down by catalase enzyme present in human cells.
 Effective in disinfection of inanimate objects.  Sporicidal at higher temperatures.
 Used by food industry and to disinfect contact lenses.
C. Benzoyl Peroxide:
 Used in acne medications.
6. Peracetic Acid :
 One of the most effective liquid sporicides available.
 Sterilant : - Kills bacteria and fungi in less than 5 minutes.
 Kills endospores and viruses within 30 minutes.
 Used widely in disinfection of food and medical instruments because it does not leave toxic residues.
6. Aldehydes:
 Highly effective molecule that combine with nucleic acids and protein and inactivate them, probably by
crosslinking and alkylating molecules.
 They are sporicidal and can be used as chemical sterilant.
 Formaldehyde ,which is available as a 37% solution in water(formalin),denature proteins and nucleic
acids.
 Glutaraldehyde is used to disinfect hospital ,laboratory equipment, respiratory therapy equipment.
MECHANICAL METHODS
1. Filtration:
 Mechanical means of removing microorganisms.
The liquid or gas is passed through a filter with
pores small enough to prevent passage of microbes.
This method can be used for substances that are
sensitive to heat.
 HEPA filters: removes at least 99.97% of airborne
particles 0.3 micrometers (µm) in diameter.
 ULPA filter: remove from the air at least 99.999%
of dust, pollen, mold, bacteria and any airborne particles with a size of 120 nanometers or larger.
2. Osmotic pressure:
 The use of high concentrations of salts and sugars in foods is used to increase the osmotic pressure and
create a hypertonic environment.
 The addition of salt to meat creates a solute-rich environment where osmotic pressure draws water out
of microorganisms, thereby retarding their growth.
Note: Staphylococci spp, Yeast are resistant to high osmotic pressures.

17
 TA SLIDES BOOK SERIES
Importance of control of microbial growth
 Control of microbial growth means to kill or inhibit the growth microorganisms. Control of growth
usually involves the use of physical or chemical agents which either kill or prevent the growth of
microorganisms.
 The control of microbial growth is necessary in many practical situations, and significant advances in
medicine, agriculture, and food science have been made through the study of microbiology.
Microbial control is very important in the following different aspects.
 Controls infectious diseases.  Minimizes cross and nosocomial infections
 Prevents surgical complications  Decreases morbidity and mortality
 Prevents food from spoilage  Food can be preserved for months
 Improves and ensures the safety of agricultural products
 Ensures the provision of safe drinking water to public
ANTIBIOTICS
“ANTIBIOTIC” is derived from antibiosis which means “ against life”
 Antibiotics are chemical substances produced from various
microorganisms (bacteria and fungi) that kill or inhibit the growth
of other microorganisms.
 An antibiotic is an imprecisely term, generally used to describe any
compound (natural or synthetic) that inhibit the growth of, or actively
kills, microorganisms.
 Antibiotics differ by mode of action
 Bacteriostatic compounds inhibit the growth of bacteria.
 Bactericidal compounds directly kill the bacteria. BACTERIOSTATIC VS BACTERICIDAL
ANTIMICROBIAL THERAPY
 An antimicrobial therapy kills or inhibits the growth of microorganisms such as bacteria, fungi, or
protozoans.
 Therapies that kill microorganisms are called microbiocidal therapies
 Therapies that only inhibit the growth of microorganisms are called microbiostatic therapies.
 Empiric
– Infecting organism(s) not yet identified – More “broad spectrum”
 Definitive
– Organism(s) identified and specific therapy chosen – More “narrow” spectrum
 Prophylactic or preventive
– Prevent an initial infection or its recurrence after infection
HISTORY OF ANTIBIOTICS
 Before penicillin introduction there was no effective treatment for treating infections.
 In 1928 penicillin, the first true antibiotic, was discovered by Alexander Fleming, Professor of
Bacteriology at St. Mary's Hospital in London.
 Alexander Fleming was a bit disorderly in his work. He left his petridishes uncovered. Upon returning
from holidays he noticed that a fungus, Penicillium notatum, had contaminated
a culture plate of Staphylococcus bacteria. The fungus had created bacteria-free
zones wherever
it grew on the plate.
 Fleming isolated and grew the mould in pure culture. He found that P.
notatum proved extremely effective even at very low concentrations,
preventing Staphylococcus growth even when diluted 800 times.
 Fleming published his findings in the British Journal of Experimental
Pathology in June 1929.
BACTERIA AND ANTIBIOTICS
 Bacteria are microorganisms that invade the human body through many routes like respiratory, GI, and
skin.
18
 TA SLIDES BOOK SERIES
 Human immune response is activated as the body tries to rid itself of bacteria, classic signs of
inflammation (e.g. swelling, heat, redness, and pain), fever begin to show up.
 The goal of antibiotic therapy is to decrease the population of invading bacteria to a point at which the
human immune system can effectively deal with the invader.
GRAM POSITIVE VS GRAM NEGATIVE BACTERIA
 Most bacteria can be broadly classified as Gram +ve or Gram -ve.
 Gram +ve bacteria have cell walls composed of thick layers of peptidoglycan.
 Gram +ve cells stain purple when subjected to a Gram stain procedure.
 Gram -ve bacteria have cell walls with a thin layer of peptidoglycan. The cell wall also includes an outer
membrane with lipopolysaccharide (LPS) molecules attached.
 Gram -ve bacteria stain pink when subjected to a Gram stain procedure.
 While both Gram +ve and Gram -ve bacteria produce exotoxins, only Gram -ve bacteria produce
endotoxins.
GRAM POSITIVE VS GRAM OVERVIEW OF BACTERIAL INFECTIONS
NEGATIVE CELL WALL

SOME IMPORTANT TERMS

 Fumigation: Means sterilization by exposure of vapors. O.T is sterilized by formaldehyde vapors.
 Isolation: Means separation of an infected person from healthy person to prevent spread of disease.
 Culture media: The media (petri dish) for cultivation (culturing) of microorganism for diagnostic and
drug sensitivity.
 Phagocytes: A type of cell within the body capable of engulfing and absorbing bacteria and other small
cell and particle (Eating cells) E.g. WBC eat microbes.
 Pathology: Pathology is the study of disease, especially of structural abnormalities produced by disease.
 Spore: An oval body formed (in bacteria) a rounded resistant form adopted by a bacterial cell in adverse
condition.
 Parenteral: Means administration of drug or nutrient by injection or infusion.
 Immunity: Means process of recognition and eliminating foreign antigen.
 Antigen: An substance that causes immune system to produce antibody against disease.
 Antibody: Antibodies are proteins that protect you when an unwanted substance enters your body
produced by your immune system.
 Nosocomial Infection: Hospital acquired infection is called nosocomial infection.
 Bacteremia: Presence of bacteria in the circulating blood .
 Bacteriuria: Presence of bacteria in urine.
19
 TA SLIDES BOOK SERIES
 Septicemia: Invasion of the bloodstream by virulent bacteria.
 Pus: Thick yellow semi liquid substance consisting dead WBCs bacterial cell and tissue fluid
 Virulence: Power of microbes to produce disease .
 Caryokinesis: Division of a cell nucleus during mitosis.
 Cytokinesis: This is division of cytoplasm.
 Chemotherapy: It means using chemical agents that are selectively toxic to the causative agent of the
disease, such as a microorganisms or malignant cells.
 Chemotherapy means the use of drugs to eradicate micro-organisms and parasitic worms or malignant
cells in the body is called chemotherapy
 Antimicrobial drugs: Drugs that are used to treat infections with micro organisms are known as
antimicrobial drugs.
 Antibiotic resistance: Antibiotic resistance is the ability of bacteria/fungi to resist the effects of an
antibiotic
BROAD SPECTRUM VS NARROW SPECTRUM SELECTION OF ANTIMICROBIAL AGENTS
 Selection of the most appropriate antimicrobial
agent requires knowing
A) the organism’s identity,
B) B) the organism’s susceptibility to a particular
C) agent,
D) C) the site of the infection,
E) D) patient factors,
F) E) the safety of the agent, and
G) F) the cost of therapy.

SPECTRUM OF ANTIBIOTICS
 Narrow-Spectrum antibiotics: A Narrow spectrum antibiotic is antibiotics that is only able to kill or
inhibit limited species of bacteria.
 Extended-Spectrum: Ampicillin acts against gram positive and some gram negative bacteria.
 Broad-Spectrum Antibiotics: Tetracycline and chloramphenicol affect a wide variety of microbial
species.
NARROW SPECTRUM BROAD SPECTRUM

WHAT IS ANTIMICROBIAL RESISTANCE

 Antimicrobial resistance is the ability of microbes to resist the effects of drugs in same dosage. When the
drug loose the ability to either kill or inhibit the growth of microbes and the microbes gain the ability to
survive in the presence of drug to which they were previously susceptible this is called resistance.
INTRINSIC RESISTANCE
 Intrinsic resistance is the innate ability of a bacterial species to resist activity of a particular antimicrobial
drug. This can also be called “insensitivity” since it occurs in organisms that have never been susceptible
to that particular drug.
20
 TA SLIDES BOOK SERIES
 Lack of affinity of the drug for the bacterial target:
for example penicillin's are not effective against mycobacterium tuberculosis, as the later does not contain
peptidoglycan in cell wall.
 Inaccessibility of the drug into the bacterial cell:
For example: Gram -ve bacteria are naturally resistant to vancomycin and penicillin G/V. Because of
inability to penetrate outer membrane.
ACQUIRED RESISTANCE
 Acquired resistance means when the microbes gains the ability to grow in the presence of a drug.
Acquired resistance develops when micro organisms no longer respond to a drug to which they were
previously susceptible.
 β-Lactamase activity: This family of enzymes hydrolyzes the cyclic amide bond of the β-lactam ring,
which results in loss of bactericidal activity
 Altered PBPs: Modified PBPs have a lower affinity for β-lactam Antibiotics
HOW TO PREVENT ANTIBIOTIC RESISITANCE

ANTIMICROBIAL DRUGS
Common microbes causing diseases:
 Bacteria......anti bacterial  Viruses……anti viral
 Fungi……...anti fungal  Protozoa ….antiprotozoal
SENSITIVITY IN MICROBIOLOGY
 Sensitivity analysis determines the effectiveness of antibiotics against microorganisms (germs) such as
bacteria that have been isolated from cultures.
 An antibiotic sensitivity (or susceptibility) test is done to help choose the antibiotic that will be most
effective against the specific types of bacteria or fungus infecting an individual person.
ANTIBACTERIAL AGENTS AND THEIR MODE OF ACTION CLASSIFICATION OF ANTIBIOTICS

21
 TA SLIDES BOOK SERIES
ANTIVIRAL DRUGS ANTI PROTOZOAL DRUGS

22
 TA SLIDES BOOK SERIES
Hypersensitivity
Introduction
 Hypersensitivity (Immunological reaction) refer to:
 Undesirable immune reaction produced by the normal immune system.
 Hypersensitivity reaction: When an immune response result in exaggerated OR in appropriate
reactions harmful to the host the term hypersensitivity OR allergy used.
 Hypersensitivity occurs when a sensitized person re-exposes to the allergen.
Types of Reaction

Type 1(Anaphylactic) Reaction

 Basic Characteristic Features:
 Type I,II,III are antibody mediated hypersensitivity and type IV is cell mediated hypersensitivity
reaction.
 Type I hypersensitivity quickly developing (2—30 min) after contact of Ag (allergen) with Ab after
previous exposition.
 IgE are involved, IgE produced in response to an antigen like insect venom or plant pollen grains, bind
to the surface of mast cells and basophils.
 Type I hypersensitivity also called Allergy.
 Involve chemical mediators.
 These all are hole marks of type I hypersensitivity reaction.
 Local reaction - skin or mucosa
 Bee sting, food allergy, hay fever (pollinosis), asthma, urticaria (hives)
 Systemic reaction ( Anaphylactic shock)
 Parenteral administration of Ag (e.g. penicillin)
 Within minutes-- itching, redding of skin, breathing problems, abdominal pain, vomiting, diarrhea.
 During several min -- death may occur due to collapse of circulation (drop in blood pressure)
Chemical Mediators of Type I Hypersensitivity Reaction
 Histamine (Most potent chemical mediator):
• Histamine causes smooth contraction and smooth present in our airway
• Constriction of smooth muscles, Bronchiole constriction .
 Slow releasing substance of anaphylaxes basically Leukotrienes.
 Eosinophilic chemotactic factor.  Serotonin
 Thromboxane A2  Platelet activating factor

23
PAGE NO
 TA SLIDES BOOK SERIES
These are six chemical mediators of type I hypersensitivity reaction.
Function:
 Smooth muscles Contraction  Increase vascular permeability
Mechanism(Two phases)
Immediate phase:
 When any antigen /allergen enter into the host and initiate the production of IgE antibody. These IgE
antibody bind to their surface of mast cell and basophil.
 Second exposure of the same allergen bind to the combination mast cell and IgE.
 These three combination allergen,IgE,Mast cell and basophil lead to the production of certain
mediator(chemical substances) from the granules.
 These granules are present in the mast cell and basophil in the preformed stage(like mediator
serotonin etc).this immediate phase complete 01 to 02 minutes.
Late phase:
 It start 5-6 minutes later.it mainly involved into leukotriene's.
 Mast cell and basophil both are destroyed.
 They burst open and they produce leukotriene and other chemical mediator outside them.
 At this stage type I hypersensitivity reaction is called anaphylaxes.

What is the sequence of events in an IgE- mediated hypersensitive response?

1. The plasma cells secrete IgE.
2. These IgE bind to Fc receptors on sensitized mast cells and blood basophils.
3. When the allergen appears again (usually a few weeks after the first exposure), it cross-links the mIgEs
and causes degranulation, releasing granules.
4. Mediators within these granules act on the
surrounding tissues such as smooth muscle, small
blood vessels, and mucous glands.
Common Allergen associated with Type I
Hypersensitivity

Clinical Sign and Symptom of Type I Hypersensitivity

 Skin- include uticaria,Dermatitis
 Lung- include bronchiole asthma, which is characterized by narrow of the airway.
24
PAGE NO
 TA SLIDES BOOK SERIES
 Nose- rhinitis  Eyes- Conjunctivitis
 GIT- involved allergic vomiting ,diarrhea
Atopy:
 Ability of any individual host to produce raised level of IgE antibody in the body.
 Increase tendency IL-4 lead to increase the production of IgE.
 IL-4 is produced by Th-1.
How to detect Type I Hypersensitivity
 Previous history ,clinical sign and symptoms
 Investigation Lab test:
1. RAST: stand for radioallergo sorbent test: In this test we just measure the amount of IgE in Nano grams
toward a specific antigen.
2. R.I.S.T: stand for radioimmuno sorbent test: In this test we also measure the amount of IgE not toward
the specific allergen. In this test we measure the total concentration of IgE.
Treatment for Type I Hypersensitivity
Pharmacotherapy :
 Drugs.  Non-steroidal anti-inflammatories
 Antihistamines block histamine receptors.  Steroids
 Theophylline OR epinephrine -prolongs or increases cAMP levels in mast cells which inhibits
degranulation.
Immunotherapy :
 Desensitization (hyposensitization) also known as allergy shots.
 Repeated injections of allergen to reduce the IgE on Mast cells and produce IgG.
Type II (Cytotoxic) reaction
Basic Characteristic Features:
 Also called antibody mediated Cytotoxic hypersensitivity.
 Two basic characteristic antibody mediated and cytotoxic reaction. These are essential component of
type II hypersensitivity.
 Type II hypersensitivity involves igG or igM antibody mediated not IgE are involved.
 IgM or IgG immunoglobulin react with cell surface antigen to activate the complement system and
produce direct damage of the cell surface.
Basic Characteristic Features:
 Complement system activated.
 Complement system actually composed of 20 or more different type of proteins forming membrane
attack complex. This membrane attack complex, it is responsible for cell destroying component of type II
hypersensitivity (membrane attack complex,C5,C6,C7,C8).
 Combination of these complement protein (C,5,6,7,8) it is actually responsible to breakdown the host
cell.
 Transfusion reaction and hemolytic
disease of the newborn are example of
type II hypersensitivity.
Hemolytic Disease of the Newborn
 This is where maternal IgG antibodies
specific for fetal blood group antigens
cross the placenta and destroy fetal
RBC’s.
 Erythroblastosis fetalis-severe
hemolytic disease of newborns.
 Most commonly develops when an
Rh+ fetus expresses an Rh antigen on it’s
blood that and Rh-mother doesn’t
recognize.
Erythroblastosis fetalis
25
PAGE NO
 TA SLIDES BOOK SERIES
• During the 1st pregnancy small amounts of fetal blood pass through the placenta but not enough to
induce a responses
• During delivery larger amounts of fetal blood cross the placenta causing an activation of B-cells that are
Rh specific thus leading to memory B-cells (anti-Rh antibodies)
• The IgM antibody clears the Rh+ cells from the mother
• In subsequent pregnancies with an Rh+ fetus, the Rh+ RBC cross the placenta activating the memory
B-cells
• These in turn cross the placenta and damage the fetal RBC because they are seen as “foreign”
• This type of reaction can be prevented by administering antibodies against the Rh antigen within 25-48
hours after the 1st delivery
• Rhogam-is the antibody that is injected – it will bind to the fetal RBC that enter the mother’s circulation
and facilitate the clearance of them before B-cell activation – In subsequent pregnancies the mother is
unlikely to produce IgG anti-Rh antibodies – If the mother doesn’t receive this injection there are other
ways to treat this, depending on the severity
Drug-Induced Hemolytic Anemia
 This is where certain antibiotics can absorb nonspecifically to the proteins on RBC membranes
 Examples: penicillin, streptomycin.
 When drug is withdrawn the hemolytic anemia disappears.
 Drug reactions
 Drug binds to Rbc surface and antibody against drug binds and causes lysis of Rbcs.
Clinical Example
ABO blood group mismatch:
 In this case antibody present in the host blood, they react against the newly entering Rbc, which have
never antigen on their surface as a result these antibody activate the complement and there is destruction
of the new blood which is donated to the host.Compare Type I and Type II hypersensitivity
Type I Type II
Involved IgE Involved IgG,IgM
Chemical mediator are involved like Involved Complement system
histamine,Seotonin
Allergic reaction No allergic reaction No
involvement of complement system Involve complement system
Type III (Immune Complex ) Reaction
Basic Characteristic Features:
 Formation of Ag- Ab complexes (immune complexes) deposit formation.
 Activation of complement system like C5a,C3a complement protein activation.
 Chronic inflammation.
 The reaction may take 3-10 hours after exposure to the antigen .
 Large amounts of immune complexes can lead to tissue damage (Type III HPS reaction).
 These component are mandatory in type III HSR.
Immune complex deposits
 Combination of antigen and antibody. This combination is called immune complex, and this
combination is deposited at different body site,e.g these immune complex may be deposit at the kidney,
under the skin, lungs, joints, etc. this is called immune complex deposits.
Complement activation
 Whenever the antigen forms immune complex with the antibody, there is activation of complement
system.
 Specially activate (C5a,C3a) they cause inflammatory process which is the 3rd component.
Chronic Inflammation
 Chronic inflammatory cells, specially neutrophil they start to accumulate at the site of the injury.
 How the injury caused, because deposition of immune complex deposits and also complement system
activate.

26
PAGE NO
 TA SLIDES BOOK SERIES
 Neutrophil are released, and complement system is activated (C5a,C3a, and neutrophil ,they release
protolytic enzyme, and these enzyme contribute toward the injury as a response type III hypersensitivity
reaction start.
Localized Type III Reactions:
1. Injection of an Antigen:
• Can lead to an acute Arthus reaction within 4-8
hours
• Localized tissue and vascular damage result from
accumulation of fluid (edema) and RBC (erythema)
• Severity can vary from mild swelling to redness to
tissue necrosis
2. Insect bite:
• May first have a rapid type I reaction
• Some 4-8 hours later a typical Arthus reaction
Develops
Generalized Type III Reactions:
• Large amounts of antigens enter the blood stream
and bind to antibody, circulation immune
complexes can form
• These can’t be cleared by phagocytosis and can
cause tissue damaging Type III reactions
• Serum Sickness-type III hypersensitivity reaction that develops when antigen is intravenously
administered resulting in formation of large amounts antigen-antibody complexes and the deposition in
tissue
• Other conditions caused by Type III-
1. Autoimmune Diseases
• Systematic lupus erythematosus • Rheumatoid arthritis
Clinical Sign and Symptom of Type III Hypersensitivity reaction
 Rheumatoid arthritis: In this disease Rheumatoid factor leads to the formation of antibody, and this
combination of antibody + Rheumatoid factor, they are deposited in the synovial joint and leads to the
destruction of joint spaces as a result patient feel joint pain.
 Systematic lupus erythematosus: It is multisystem disease, it involve different system of the body
like,skin,lungs,kidney,etc. What happened, antibody are formed against the DNA and this leads to
destruction of different tissue of the host.
 Glomerulonephritis.  Good pasture syndrome.
Compare between Type II and Type III hypersensitivity
Type II Type III
In type II hypersensitivity antibody are formed In type III hypersensitivity antibody are formed against the
against tissue specific antigen. circulating antigen.
No Immune complex deposit formation Immune complex deposit formation.
Type IV (cell mediated) Reaction
Basic Characteristic Features:
 cell mediated hypersensitivity or delayed type hypersensitivity. What is delayed type hypersensitivity
(DTH)?
 A hypersensitive response mediated by sensitized TDTH cells, which release various cytokines and
chemokines.
 Generally occurs 2-3 days after TDTH cells interact with antigen.
 T cell are involved both T helper cell CD4,cytotoxic T cell CD8.
 Macrophage are involved.  Dendritic cell are involved (present epidermis of the skin).
 cytokines mainly (IL2,IL4,).
Basic Characteristic Features:
 There is no role of neutrophil in type IV hypersensitivity reaction.
27
PAGE NO
 TA SLIDES BOOK SERIES
 Mast cell are involved in type IV hypersensitivity reaction but this involvement of mast cell is very
limited.
 In type I mast cell are suddenly destabilized here in this case mast cell are not suddenly destabilized,
they release chemical substances which recruit other cells of inflammation at the site of type IV
hypersensitivity reaction.

Different Variants of type IV hypersensitivity Reaction

Contact hypersensitivity:
In contact hypersensitivity CD8,cytotoxic T lymphocyte are involved in a major component and they
destroy the target cell.
Tuberculin test:
Indicates delayed character of type IV hypersensitivity reaction. In this basis type IV hypersensitivity are
called delayed HPS. There are more involvement of macrophages in tuberculin testing along with CD4
helper cells.
Granuloma hypersensitivity:
In this case macrophages are transformed into joint cell called Epithelioid cells . It is the combination of
lymphocyte ,fibroblast, and connective tissue ,this combination is called granuloma. This granuloma to the
whole mark of tuberculosis.
Type I Diabetes Mellitus:
This is also clinical sign of type IV hypersensitivity reaction.
Mechanism
 Antigen which enter the skin reach to the epidermis, it become attach with the carrier protein which is
already present there and this antigen here called hepton.
 This hepton combine with carrier protein in the epidermis of the skin ,now this combination called
complete antigen .
 Recognition and processing by antigen presenting cells, after entering the hepton and formation of
complete antigen recognize and process by the antigen presenting cells. These may be macrophages, these
may be dendritic cells present in the epidermis.
 Migration of APC,s to the lymps nodes where they present the complete antigen into the T
lymphocyte.
 Cytokine: now release of cytokine which stimulates proliferation of T -lymphocyte and activate
macrophages.
 These T cell activate macrophages .
 They migrate toward the epidermis and release inflammatory mediator as a result they destroy the
target cell.
Phases of the DTH Response
Sensitization phase: occurs 1-2 weeks after
primary contact with Ag What happens during this
phase?
• TH cells are activated and clonally expanded by
Ag presented together with class II MHC on an
appropriate APC, such as macrophages or
Langerhans cell (dendritic epidermal cell)
• Generally CD4+ cells of the T subtype are
activated duringH1 sensitization and designated as
TDTH cells Effector phase: occurs upon subsequent
exposure to the Ag What happens during this
phase?
• TDTH cells secrete a variety of cytokines and
chemokines, which recruit and activate
macrophages
• Macrophage activation promotes phagocytic
28
PAGE NO
 TA SLIDES BOOK SERIES
activity and increased concentration of lytic enzymes for more effective killing
• Activated macrophages are also more effective in presenting Ag and function as the primary effector cell
Detrimental Effects of DTH Response (diagram on previous page)
 The initial response of the DTH is nonspecific and often results in significant damage to healthy tissue
 In some cases, a DTH response can cause such extensive tissue damage that the response itself is
pathogenic
 Example: Mycobacterium tuberculosis – an accumulation of activated macrophages whose lysosomal
enzymes destroy healthy lung tissue.
POINT TO BE NOTED:

29
PAGE NO
INTRODUCTION
֍ From the months spent in the womb to the end of life, every individual is under constant attack from an
enormous range of potentially harmful invaders.
֍ These threats include such diverse entities as bacteria, viruses, cancer
cells, parasites and foreign (non-self) cells, e.g. in tissue transplant.
֍ The body has therefore developed a wide selection of protective
measures against the invaders.
Common Definitions
֍ Resistance: The ability of the body to resist against invaders and protect the body from getting damage.
֍ Susceptibility: The meaning of SUSCEPTIBILITY is the quality or state of being susceptible; especially :
lack of ability to resist some extraneous agents like microbes. Or lack of resistance is called susceptibility.
֍ Non Specific resistance: Non specific resistance is the defense of our body from any kinds of the
pathogens. It includes skin and mucous membrane, phagocytosis, inflammation, fever, production of
antimicrobial substances.
֍ Specific resistance: the ability of the body to defend itself against specific invading agents such as
bacteria, toxins, viruses and foreign tissues.Common Definitions
֍ Innate resistance: The inherited or by birth ability of the body to resists against invaders such as
bacteria virus, foreign body cells etc.
֍ Immunity: Immunity is defined as the body's ability to protect itself from an infectious disease. When
you are immune to a disease, your immune system can fight off infection from it.Common Definitions
֍ Antigens: Any substance that causes the body to make an immune response against that substance.
Antigens include toxins, chemicals, bacteria, viruses, or other substances that come from outside the body.
֍ Antibody: An antibody is a protein produced by the body's immune system when it detects
harmfulsubstances, called antigens.
Non-specific vs Specific defence mechanism Line of defence

Line of defence Line of defence

Non-specific resistance
֍ These are the general defense; they prevent entry and minimize further passage of microbes and other
foreign material into the body. Also destroy pathogens inside the body by various mechanisms.

30
֍ There are five main non-specific defense mechanisms:
• Defense at body surfaces
• Phagocytosis (1 st line of defenses)
• Natural antimicrobial substances (2nd line of defenses)
• The inflammatory response
• Immunological surveillance. (3rd line of defenses)
Non-specific resistance
1.Defense at body surfaces:
֍ Healthy, intact skin and mucous membranes provide an efficient physical barrier protecting the body’s
exposed surfaces.
֍ Sebum and sweat secreted onto the skin surface contain antibacterial and antifungal substances.
֍ Epithelial membranes lining body cavities and passageways exposed to the external environment.
Epithelia produce antibacterial secretions, often acidic, containing antibodies and enzymes, as well as
sticky mucus for trapping passing microbes.Non-specific resistance
2.Defense at body surfaces:
֍ Hairs in the nose act as a coarse filter, and the sweeping action of cilia in the respiratory tract moves
mucus and inhaled foreign materials towards the throat. Then it is coughed up (expectorated) or
swallowed.
֍ The one-way flow of urine from the bladder minimizes the risk of infection ascending through the
urethra into the bladder.
֍ In the female, the acidity of vaginal secretions discourages microbial growth.Non-specific resistance
3.Phagocytosis:
֍ Also Known as the process of cell
eating. ֍ Phagocytic defense cells
such as macrophages and neutrophils
are the body’s first line of cellular
defense. ֍ They actively migrate to
sites of inflammation and infection.
֍ Phagocytes attack and engulf their
targets. ֍ They digest and destroy
foreign cells, damaged body cells and
debris. ֍ Macrophage stimulate
T-lymphocytes and activate the
U- immune responses.
4.Natural antimicrobial substances:
֍ Hydrochloric acid: Found in stomach------Kill majority of the ingested microbes
֍ Lysozyme: Found in saliva, tears and other body secretions— destroy bacterial C-wall.
֍ Saliva: Produced by salivary glands– contain antibodies and lysozyme– prevent dental decaying by
disrupting microbes and washing mouth,
֍ Interferon: These are chemicals produced by T-lymphocytes, macrophages and other body cells
invaded by viruses--- prevent viral replication within infected cells & the spread of virus to healthy cells.
֍ Microbial Antagonism: one microorganism kill or inhibit the growth of another microorganism
Non-specific resistance
5.The inflammatory response:
֍ This is the physiological response to tissue damage and is accompanied by a characteristic series of local
changes.
֍ Its purpose is protective: to isolate, inactivate and remove both the causative agent and damaged tissue,
so that healing can take place.
֍ The cardinal signs of inflammation are redness, heat, swelling and pain.
֍ Inflammatory conditions are recognized by their Latin suffix ‘-itis’; for example, appendicitis is
inflammation of the appendix and laryngitis is inflammation of the larynx.

31
6.Immunological surveillance:
֍ A population of lymphocytes, called natural killer (NK) cells, constantly patrol the body searching for
abnormal cells.
֍ Cells that have been infected with a virus, or mutated cells that might become malignant, frequently
display unusual markers on their cell membranes, which are recognized by NK cells.
֍ Having detected an abnormal cell, the NK cell immediately kills it.
Specific resistance ( Acquired Immunity)
1.Specific resistance:
֍ the ability of the body to defend itself against specific invading agents such as
bacteria, toxins, viruses and foreign tissues.
2.Immunity:
֍ Immunity is a biological term that describe a state of having sufficient biological defenses to avoid
infections, diseases, or other biological invasions.
֍ The ability of the body to resists harmful microbes from entering the body.
Types of immunity
1.Innate Immunity ( Non-specific):
֍ Natural resistance or by birth
resistance. ֍ It include the roles of:
 Physical Barriers: eg. Skin & Mucus
membranes  Cellular system: eg.
Phagocytes like macrophage
 Circulating proteins and enzymes:
eg. immunoglobulin, Lysozyme
 Antimicrobial substances: eg.
interferon, HCL, Bile vaginal
discharge, saliva, tears
 Microbial Antagonism: eg. Normal Flora
 Inflammatory responses: eg fever, pain, swelling, heat
Innate immunity

2.Adaptive or acquired Immunity (specific):

֍ It develops gradually through a person’s lifetime. When exposed to a disease or
when immunized against it with the vaccine, our body develops adaptive
immunity. It can be sub-divided by how the immunity was acquired:
֍ Sub-divided into two classes:
1. Natural acquired immunity
 Naturally acquired PASSIVE immunity  Naturally acquired ACTIVE immunity

32
2. Artificial acquired immunity
 Artificially acquired ACTIVE immunity  Artificially acquired PASSIVE immunity

Humoral Immunity
֍ Humoral immune is a response
immunity that is mediated through
antibodies. It is the core defense
mechanism against extracellular
microbes. It responds to pathogens
quickly.
֍ It triggers the B cells to produce
antibodies that bind with foreign
antigen then neutralized them
through phagocytosis.
Cell mediated immunity
֍ Cell-mediated immunity is
the immune response that
identifies and destroys
infected cells to prevent the
spread of viruses or bacteria.
֍ The immune response
trigger development of T cells
which are released into the
thymus and circulate between
the peripheral lymphoid tissue
and the blood.
֍ The process helps to identify
foreign antigens and get rid of
them immediately. The immune response it quite stronger than humoral immune.
Humoral vs CMI
Basic Terms Humoral Immunity Cell-mediated Immunity
Onset Rapid Delay
Activation Differentiation of plasma B
cells and secretion of antibodies Secretion of cytokines
Main cells B-cells which are generated and T-cells generated in the bone
mature in the bone marrow marrow and mature in the thymus
Pathogens Protect against extracellular Protect against intracellular
Pathogens pathogens
Meaning Antibodies are formed Antibodies are not formed
Antigen Detection Antibodies Receptors
33
Basic Terms Humoral Immunity Cell-mediated Immunity
Examples of cells Only TH cells CD4+ and CD8+ cells
Cancer cells Cannot be eliminated Can easily be eliminated
Rejection Involve in early graft rejection Participate in organ transplant
rejection
Core function It protects against extracellular It protects against intracellular
bacterial and viral pathogens. bacterial and viral pathogens.

CMIR AND AMR Difference b/w Humoral & CMI

Antigen

34
 Antibodies Y- Shape of antibody

How antibodies bind to an

Y-shape of antibody antigen

TYPES Functions of Antibodies

35
 TA SLIDES BOOK SERIES
UNIT 04 Asepsis & Concept of Asepsis
Some Important Terms
 Infection:
An infection is the invasion of body tissue by disease- causing microorganism, their multiplication and the
reaction of body tissue to these microorganism and the toxins that they produce.
 Inflammation :
Inflammation is the response of body to infection or injury which is characterized by swelling, heat,
redness and pain.
 Pathogenicity:
Pathogenicity is the ability of a pathogen to produce an infectious disease in an organism.
Sepsis
 Sepsis is a condition in which the body is fighting a severe infection that has spread via the bloodstream.
 If a patient become septic, they will likely have low blood pressure leading to poor circulation and lack of
perfusion of vital tissue and organ.
 This condition is termed (shock) and is something referred to as septic shock.
Definition of Asepsis
 The term asepsis means the absence of disease-producing microorganisms.
 Reducing the number of microbes to an irreducible number.
 Sum total of the effort to keep the patients environment free from contamination and the patient free
from colonization.
Concept of Asepsis
 The Nurse’s efforts to minimize the onset & spread of infection are based on the principles of aseptic
technique.
 Aseptic technique is an effort to keep the client as free from exposure to infection-causing pathogens as
possible.
 Aseptic technique is the effort taken to keep the patient as free
from hospital micro-organisms as possible.
 This can be achieved by ensuring that only sterile equipment &
fluids are used during invasive medical & nursing procedures.
Types of Asepsis
 There are two types of asepsis:
Medical asepsis & Surgical asepsis.
 Medical Asepsis :also known as cleaning technique.
 Surgical Asepsis : refers to sterile technique.
Common Aseptic Procedures
 Use of temperature, pressure and radiation.  Hand washing .
 Use of disinfectant and antiseptic.  Use of personal protective equipment.(PPEs)
 Application of autoclave and sterilization in clinical environments.
 Isolation due to communicable disease.  Quarantine .
Medical Asepsis
 Definition:
 The practice of medical Asepsis helps to prevent infectious organisms and to maintain an environment
free from contamination.
 The technique used to maintain medical asepsis include hand washing, gowning and wearing facial
masks when appropriate.
 All the measures aimed at reducing the number or spread of microorganism.
 During daily routine care, the nurse uses basic medical aseptic techniques
to break the infection chain.
 Example of medical asepsis are changing client’s bed linen daily, hand
washing, barrier techniques, & routine environmental cleaning.
 Follow Isolation technique as appropriate.
36
 TA SLIDES BOOK SERIES
 Clients with high susceptibility to infection require special precautions to prevent exposure to
pathogens.
 In medical asepsis, an area or object is considered contaminated only if it is suspected of containing
pathogen .
 Hand Washing is essential component of maintaining medical asepsis.
Principles of Medical Asepsis
 When the body is penetrated, and natural barriers such as the skin is bypassed, the patient is susceptible
to any microbes that might enter.
 Even though intact skin is a good barriers against microbial contamination, a patient can become
colonized with microbes if appropriate precautions are not taken. Components of maintaining
 All body fluids from any patient is considered contaminated. Medical Asepsis
 A clean environment reduces microorganisms.
 Personal protective equipment serve as barrier to microbial
transmission.
 The health care team and the environment can be a source of
contamination for the patient.
 Assess each patient to determine if he has an infectious process.
 Choose the barrier appropriate to the infectious process.
 Isolate the disease not the patient.
Surgical Asepsis
 Definition:
 Refers to those practices that keep an area, or object free of all
microorganisms and the spores.
 Sterile technique is used to prevent the introduction or spread
of pathogen from the environment into the patient.
 Refers to the procedure used to keep the object or areas sterile from microorganism.
 Sterilization destroys all microorganisms & their spores.
 Surgical asepsis demands the highest level of aseptic technique & requires that all areas be kept as free
as possible of infectious micro-organisms.
 These techniques can be practiced by nurses in the OR
(surgical incision) or at the bedside (e.g., inserting IV or
urinary catheter & reapplying sterile dressings) where
sterile instruments & supplies are used.
 In surgical asepsis, an area or object may be considered
contaminated if touched by an object that is not sterile.
 The Nurse working with a sterile field or with sterile
equipment must understand that the slightest break in technique
results in contamination.
 A Nurse in an operating room follows a series of steps to
maintain sterile techniques, including applying a mask, protective
eyewear, and a cap; performing a surgical hand washing; &
applying a sterile gown & gloves.
 Effectiveness of aseptic practices depends on the nurse’s
conscientiousness & consistency in using effective aseptic techniques.
Principles of Surgical Asepsis
 Always face the sterile field .Do not turn back or side on a sterile field.
Sterile objects which are out of vision are considered questionable.
 Keep sterile equipment above your waist level or above table level.
 Do not speak, sneeze and cough over a sterile field. Never reach across
sterile field.
 Keep the unsterile objects away from the sterile field.
37
 TA SLIDES BOOK SERIES
 Keep the sterile field dry.
 Once a sterile item is opened or uncovered, it become contaminated.
 The edge of the sterile field is considered unsterile.
 Handle liquids cautiously near the sterile field or prevent drapes or
wrappers from becoming wet.
 Avoid sweeping and dusting when the sterile objects are opened.
 Put on mask, wash hands, put on gowns and gloves before handling
sterile supplies.
 Never assume that object is sterile . Always check the sterility expiration date.
Aseptic Practices

CONTROL OR ELIMINATION OF INFECTIOUS AGENTS

 Cleansing:
 Is the removal of all foreign materials such as soil & organic material from objects.
 Generally, cleansing involves use of water & mechanical action with or without detergents.
- Disposable object has to be discarded.
- Reusable objects must be cleansed thoroughly before disinfection
& sterilization.
 When cleaning equipment that is soiled by organic material such as blood, fecal matter, mucus or pus,
the nurse applies a mask, protective eyewear, & waterproof gloves.
 These barriers provide protection from infectious organisms.
 A brush, detergent or soap are needed for cleaning.
Disinfection & Sterilization
 Disinfection – eliminates pathogenic organisms on inanimate objects
with the exception of bacterial spore. Noninfectious microorganisms may
or may not be killed.
 This process can be carried out by two methods:
 Physical method
 Chemical method
Sterilization – is the process of eliminating and
destroying all microorganisms, including spores
& viruses.
THE PRINCIPLES OF ASEPTIC TECHNIQUE IN WOUND CARE
 Medical Hand Washing – cleaning of trolley, opening
the dressing pack, to cleaning the wound.
 Surgical Hand Washing – scrubbing in surgery
Gowns & Aprons
 Protective clothing is used to reduce bacterial spread
by contact (nurse’s uniforms become heavily
contaminated during clinical procedures.

38
 TA SLIDES BOOK SERIES
Gloves
 The purpose of wearing gloves is both to protect the hands
from contamination by micro-organisms and to prevent the
transfer of microorganisms already on the hands.
Single-use
 Irrigation devices should not be used for multi-use purposes
as there is potential for cross-infection between pts. e.g., IV
lines & buckets.
Chain of infection
 The chain of Infection describes how microorganisms are transmitted from one person or place to
another. This could be via someone's hands, on an object, through the air or bodily fluid contact.
 The 'chain of infection' is used to represent the series of steps that allow an infection to be passed from
one person to another.

1: Infectious Agents/ Germs

Infectious agents (pathogens) include not only bacteria but also viruses, fungi, and parasites. The virulence
of these pathogens depends on their number, their potency, their ability to enter and survive in the body,
and the susceptibility of the host.
2: Reservoir
 A reservoir is any person, animal, arthropod, plant, in which an infectious
agent normally lives and multiplies.
 Animate reservoirs include people, insects, birds, and other animals.
Inanimate reservoirs include soil, water, food, feces, intravenous fluid, and
equipment.
3: Portal of exit
 Portals of exit is the means by which a pathogen exits from a reservoir.
 For a human reservoir, the portal of exit can include blood, respiratory
secretions, and anything exiting from the gastrointestinal or urinary tracts.
4: Mode of transmission
 Contact transmission
Direct
Indirect
droplet
 Vector borne
 Biological
 Mechanical
 Air-born
39
 TA SLIDES BOOK SERIES
5: Portal of entry
 Infectious agents get into the body through various
portals of entry, including the mucous membranes,
non-intact skin, and the respiratory, gastrointestinal,
and genitourinary tracts. Pathogens often enter the
body of the host through the same route they exited
the reservoir, e.g., airborne pathogens from one
person’s sneeze can enter through the nose of another
person.
6: Susceptible host
 The final link in the chain of infection is a susceptible
host, someone at risk of infection. Infection does not
occur automatically when the pathogen enters the
body of a person whose immune system is functioning normally. When
a virulent pathogen enters an immune-compromised person, however,
infection generally follows.
 Whether exposure to a pathogen results in infection depends on
several factors related to the person exposed (the host), the pathogen
(the agent), and the environment.
The Spread of Infection
 An elderly patient, hospitalized with a gastrointestinal disorder caused by E-coli, was on bed rest and
require assistance for activities of daily living.
 The patient had frequent uncontrolled diarrhea stools and the nurse provided excellent care to maintain
cleanliness and comfort.
 Following one episode of cleaning the patient and changing the bed linen, the nurse immediately went
to a second patient to provide care with Foleys catheter. The nurse hands were not washed before
assisting the second patient.
 Infection Agent:
(Escherichia Coli)
 Reservoir:
Large intestine: E coli, bacteria in the large intestine of human forms the greater part of the normal
intestine flora.
 Portal of Exist:
E.Coli exited the body in faces.
 Mode of Transmission:
The nurse removed the contaminated linen from the bed. The E.Coli organism contaminated the hands of
the nurse who then provided morning care to another patient.
 Portal Of Entry:
The second patient receiving care had a foley catheter. The nurse manipulated the tubing attached to the
catheter. The E.coli organism on the nurse hands contaminated the catheter tubing and ascended to the
patients meatus and then into the urinary bladder.
 Susceptible Host:
The second patient with a foley catheter. This patient was elderly and had a chronic illness necessitating
complete bed rest. The foley catheter contaminated by the E.Coli organism provided a direct route into the
urinary bladder.
Standard Precautions
 Standard precaution define all the steps that should be taken to prevent spread of infection from person
to person when there is an anticipated contact with:
 Blood  Body fluids
 Secretions, Such as phlegm
 Excretion, such as urine and feces whether or not contain visible blood.
40
 TA SLIDES BOOK SERIES
 Non-intact skin, such as an open wound  Mucous membrane, such as the mouth cavity.
Standard Precautions Practices
 Hand washing, the most important infection control method.
 Use protective gloves.  Masks, eye protection and/ or face shield.
 Gowns.
 Proper handling of soiled patient care equipment
 Proper environmental cleaning.  Proper disposal of needles and other sharp equipment.
 Placement in a private room for patient who can not maintain appropriate
cleanliness or contain body fluids.
Transmission Based Precautions
 Transmission based precautions may be needed in addition to standard precautions for selected
patients who are known or suspected to certain infection.
 These precautions are divided into three categories that reflect the differences' in the way infections are
transmitted.
o Airborne Precautions. o Droplet Precautions.
o Contact Precautions.
Breaking the Chain of Infection

Sources of Infection
Patient flora:
• Cutaneous • Gastrointestinal • Genitourinary
• Respiratory
Contaminated hospital environment
• Instruments • Fluids • Food
• Air • Medications
Medical Personals:
Infected • Carrier
Invasive devices:
• Urinary catheter • Vascular catheter • Endotracheal tube
• Endoscope • Drainage tube
Risk factors for infection to occur
 Admission as an emergency and to the intensive care unit (ICU).
 Hospital stay longer than seven days
 Immuno suppressant medication.
 Neutropenia.  Young age or old age.
 Lack of hygiene.  Lack of vaccination.
41
 TA SLIDES BOOK SERIES
 Food safety precautions.  Not maintaining isolation / Quarantine when chronic ill.
 Safe travel.
 Placement of:
– Central venous catheter – Indwelling urinary catheter
– Endo-tracheal tube;
 Age of the patient
 Disease status.Prevention and Control of infections
Prevention and Control of infections
 Definition:
 'Identifying and reducing the risk of infections developing or
spreading’’ Infections
Prevention & Control
 Establishment of infection control program
 Cleaning and disinfection  Sterilization
 Skin decontamination  Safe disposal of hospital waste
 Prophylactic antibiotics  Isolation
 Hospital building and design  Personnel welfare
 Provision of safe blood  Surveillance
 Personal protective equipment's  Aseptic techniques
WHO 5-Movements of Hand hygiene
1. Before patient care 2. After environmental contact Hand Hygiene
3. After exposure to blood/body fluids
4. Before an aseptic task 5. After patient care
Prevention & Control
 Gowns or Plastic Aprons:
Wear any time it is likely that clothing or skin will be soiled.
 Masks:
- wear in OR / sterile area
-wear when working directly over large areas of open skin.
- wear when it is likely that nasal & oral mucous membranes .

Room Selection: Assign patient with infectious disease

to an individual OR or last on surgical list.
 Trash & Linen: Bag all soiled trash & linen securely.
- Discard according to facility policy.
- Wear gloves & protective garments when handling
soiled linen & trash.
 Housekeeping:
 Clean all rooms on regular schedule.
 Clean articles, equipment & furniture soiled with moist
body substances immediately. Wear gloves.
42
 TA SLIDES BOOK SERIES
 Laboratory Specimens:
 Handle all laboratory specimens with equal care. Special precautionary
labels are required.
 Compliance of Care Providers:
 Develop programme to ensure that health care workers comply with the
infection precautions system.
Nursing Interventions
 The knowledgeable nurse balance the principles and asepsis, standard
precautions and psychological support.
 Knowledge of the infectious agent allows the nurse to use protective measures without isolating the
patient beyond what is necessary.
 Psychological support for the patient in isolation comes in many, such as allowing an individual to
express feelings about the constraints of isolation and providing information about the purpose of barrier
techniques.
 The nurse provides psychological support through the development and maintenance of an effective
nurse patient relationship.
 Prior to leaving the room, health care workers should remove the gloves, and wash their hands with
medicated soup in addition, they may need to wear protective gowns if there is a chance of contact with
potentially infective materials such as diarrhea or wound drainage that can not be contained or if there is
likely to be extensive contact with the patient or environment.
 Patient care limits, such as stethoscope , that are used for a patient in contact precautions should not be
shared with other patients unless, they are properly cleaned and disinfected before reuse.
Remember!!!!!
 The nurse is responsible for providing the client with a safe environment.
 The nurse’s first responsibility to the client is to first do no harm.
 It is easy to forget key procedural steps or, when hurried, to take shortcuts that break aseptic
procedures. However, the nurse’s failure to be meticulous will place the client at risk for an infection that
can seriously impair recovery.
(Florence Nightingale, 1859)

43
 TA SLIDES BOOK SERIES
CONCEPT OF ISOLATION
Isolation Defined
 Separation of an infected patient from other members of the society to prevent the spread of the disease
is called isolation .
 Isolation is the separation of a patient from contact with others in order to control the spread of an
infectious or communicable disease.
 The separation of a person with infectious disease from contact with other human beings for the period
of communicability.
 According to the CDC, isolation is the act of separating a sick individual with a contagious disease from
healthy individuals without that contagious disease in order to protect the general public from exposure of a
contagious disease.
 Isolation refers to the precautions that are taken in the hospital to prevent the spread of an infectious
agent from an infected or colonized patient to susceptible person.
Need Of Isolation
 Isolation aimed controlling and preventing the spread of infection.
 Isolation precautions are designed (like-rooms/wards) to minimize the transmission of infection in the
hospital by using updated and skilled technology.
 Isolation helps to protect patients, family members, visitors and health care workers from the spread of
infection.
 To reduce the risk of spreading certain infections or antibiotic resistant germs to other patients and staff.
 To protect patients from infection if they have a weak immune system due to certain diseases.
Key points
 Microorganisms can be spread from patients to patients and patients to staff.
 Isolation precautions can reduce transmission, decreasing the spread of microbes.
 Components: hand hygiene, personal protective equipment, single rooms, ventilation, and restriction of
movement.
 Must be applied according to signs and symptoms.
 Microorganisms causing healthcare-associated infections can be spread from infected or colonized
patients to other patients and to staff.
 Appropriate isolation/precautions can reduce transmission if they are applied properly.
 An isolation/precautions policy aims to decrease the spread of infectious agents between staff and
patients to such a level that infection or colonization does not occur.
 Isolation/precautions policies have several parts:
 Hand hygiene, personal protective equipment (protective clothing and other items), single rooms with
more or less sophisticated ventilation, and restrictions for movement of patients and staff.
 Apply isolation/precautions according to signs and symptoms of the patient and do not wait for
laboratory results.
Chain of Infection
 Infection results from an interaction between an infectious agent and susceptible host.
 The interaction occurs by means of contact between the agent and the host and is affected by the
environment.
 Breaking the chain of infection by interrupting transmission is generally the best way to prevent
infections.
 The chain of infection consists of the following components: infectious agent, reservoir, portal of exit,
mode of transmission, portal of entry and susceptible host.
 Isolation precautions focuses on interrupting this chain of infection, especially transmission.
Transmission of Infections
 Contact Spread:
 Direct: from one person to another
 Indirect: contaminated equipment or surfaces
 Droplet Expelled: when sneezing or coughing; less than 2m from the source.
 A pathogen may be transmitted by a single route or it can be transmitted in several ways.
 Airborne Spread: Transmission occurs by dissemination of airborne droplet nuclei, tiny particles
containing microorganisms that remain suspended in the air for long periods.

44
 TA SLIDES BOOK SERIES
Transmission of Infections
 Direct contact occurs when microorganisms are transferred from one person to another.
 It can involve contact with blood, body fluids, excretions, or secretions.
 During patient care by the healthcare worker or a visitor or family member.
 During interactive activities between patients.
 Indirect contact involves the transfer of a microorganism through a contaminated intermediate object,
substance, or person, e.g., contaminated equipment, food, water, or supplies.
 May occur when Inadequate hand washing is performed by a care provider.
 Equipment is not cleaned, disinfected, or sterilized adequately between patients.
 Blood borne pathogens are transferred by sharps or needle-stick injuries, transfusion, or injection.
 Droplet spread: Infectious droplets that are expelled, e.g., when sneezing or coughing, are too heavy to
float in air and can be transferred less than 2m from the source of the droplets.
 Spread may be direct or indirect.
 Direct droplet transmission: Droplets reach mucous membranes or are
inhaled.
 Indirect droplet transmission: Droplets fall on to surfaces or hands and are transmitted to mucous
membranes or food.
 Indirect droplet transmission is often more efficient than direct transmission.
 Infections spread this way include the common cold, influenza, and respiratory syncytial viruses.
Airborne Infections
 These microorganisms are generated from the infected person, generally by coughing, sneezing, and
talking, or by procedures which generate secretions (suctioning, bronchoscopy, etc.) and may be inhaled by
a susceptible host.
 Airborne Respiratory Precautions are used to prevent the spread of TB (Mycobacterium Tuberculosis),
Measles, Severe Acute Respiratory Syndrome.
Standard Precautions
 Adaptation of uniform precautions for all patients regardless of their diagnosis is called Standard
(Universal) Precautions.
 Guidelines for preventing exposure to blood, body fluids, secretions, excretions, broken skin, or mucous
membranes.
 Based on the concept that body fluids from any patient can be infectious.
 Should be used on every patient.
 Use necessary PPE for protection.
Standard Precautions
I. Hand hygiene.
II. Use of personal protective equipment (e.g., gloves, gowns,
masks).
III. Safe handling of potentially contaminated
equipment/surfaces/sharp disposal in the patient environment.
IV. Respiratory hygiene/cough etiquette.
V. Safe injection practices.
VI. Use of mask.
Use of PPEs
 Gloves: Needed if coming into contact with fluids
(vomit, stool, urine, mucous, etc.) and blood.
 Gown: Needed if there will be a potential contact with
fluids or blood on clothing.
 Goggles/Face Mask/Shield: Needed if there will be a
potential contact with fluids or blood in mucous membranes
(example: patient coughing, vomiting, while providing
suctioning/mouth care, etc.)
Transmission Based Precautions
 Transmission based precautions may be needed in
addition to standard precautions for selected patients who
are known or suspected to certain infections.
45
 TA SLIDES BOOK SERIES
 These precautions are divided into three categories (contact, droplet & air born) that reflect the
differences in the way infections are transmitted.
 Some diseases may require more than one isolation category.
Types of Isolation
 Patients are isolated according to the mode of transmission of the disease.
I. Contact Isolation II. Droplet Isolation III. Respiratory Isolation
IV. Strict isolation V. Enteric Isolation VI. Protective Isolation
VII. Wound and skin Isolation
VIII. Blood Precautions IX. Discharge Precautions
Contact Precautions
 Direct and indirect contact transmission is the most frequent route of transmission of hospital acquired
infections (Nosocomial Infections).
 Direct contact transmission occurs when there is direct body-to-body transfer of microorganisms
between an infected or colonized person and a susceptible host.
 Indirect contact transmission occurs when an inanimate object, such as equipment used on the infected
person or unwashed hands, serves as a vector of transmission between the susceptible host and the infected
person.
Recommendations for Contact Precautions
 Place patient in a single room, put on a gown and gloves before entering the patient’s room.
 Remove gown and gloves just inside the door to the patient’s room and discard within the room.
 Wash hands thoroughly after glove and gown removed.
 Dedicate equipment to the isolated patient.
 If this is not possible, equipment must be disinfected before being used on another patient.
 The patient must remain in the isolation room at all times except for essential tests or procedures that
cannot be performed in the room.
 The patient must wear a cover gown and gloves when out of the room.
Recommendations for Contact Enteric Precautions
 Used for patients with C. difficile associated disease.
 The recommendations for patient care include all of the above recommendations for Contact
Precautions, but in addition, hand washing with soap and water is required when caring for patients with
C.difficile associated disease. (Clostridium difficile is not eliminated by alcohol-based hand rub products)
 Alcohol-based hand rubs may not as effective against spore-forming bacteria.
Procedure for Contact Precautions
 All steps as in STANDARD PRECAUTIONS along with:
I. GLOVES - Gloves should be worn when entering the room.
II. GOWNS - A gown should be worn for contact with the patient and with patient items.
III. PLACEMENT -The patient should be placed in a separate (isolation) room.
 If a separate room is not available, the patient may be placed in a room with another patient with an
identical infectious condition (“cohorting”).
 Appropriate signage to be placed outside the room.
 Limit the number of visitors as appropriate and family or care givers should be educated on contact
precautions and hand hygiene.
IV. TRANSPORT – Activities of the
patient may need to be limited and when
transportation is required, the following
measures should be undertaken:
 Patient’s body is contained or covered
 Avoid contact with objects and if
necessary use gloves and maintain hand
hygiene
V. PATIENT CARE EQUIPMENT -
 a) Use of dedicated patient care
equipment should be considered where
possible (e.g. stethoscope)
 b) Where this is not possible, items
46
 TA SLIDES BOOK SERIES
should be disinfected after each use.
VI. ENVIRONMENTAL MEASURES -
 a) Focus on room cleaning and disinfection (e.g., daily bed rails, over bed table, bedside commode,
lavatory surfaces in patient bath rooms, door knobs) and equipment in the immediate vicinity of the patient.
 b) Housekeeping services need to be done at the end and then change the mops, dusters, solutions.
Infections which requires Contact Precautions
 Medication Resistant Diseases:
 MRSA (Methicillin Resistant Staphylococcus Aureus)
 VRE (Vancomycin Resistant Enterococci)
 C.Diff ( clostridium difficile)
 ESBL (Extended Spectrum Beta Lactamase Producing Enzymes Inhibitors)
 MDR (Multidrug Resistant Strains).
 Scabies.  Norovirus  Rotavirus  Hepatitis A
Droplet Precautions
 Droplet transmission occurs when droplets containing microorganisms generated from the infected
person, primarily during coughing, sneezing, or talking, or during procedures which generate secretions
(suctioning, bronchoscopy, etc.) are propelled a short distance (less than 3 meters) and deposited on the
susceptible host’s eyes, nose, or mouth.
 These droplets are relatively large and do not remain suspended in the air, therefore negative pressure
rooms are not necessary.
Recommendations for Droplet Precautions include:
 Place the patient in a single room.
 Wash hands thoroughly before entering and after exiting the room.
 Put on an isolation mask before entering the patient’s room.
 Patients may not leave the room except for essential tests or procedures that cannot be performed in the
isolation room.
 The patient should wear an isolation mask when out of the
room.
Infections that requires Droplet Precautions:
 Influenza (flu)
 Pertussis (whooping cough)-(Bordetella Pertussis)
 Mumps  Diphtheria (pharyngeal)
 German Measles (Rubella)  Streptococcal Pharyngitis
 Meningitis  Pneumonia
 Respiratory illnesses, such as those caused by coronavirus
infections
Airborne Contact Precautions
 Airborne Contact Precautions are used to prevent transmission of varicella (chickenpox, disseminated
herpes zoster, or localized zoster in an immune compromised patient).
 Recommendations for Airborne Contact Precautions include:
 A negative pressure, single room is required.
 The door to the room must remain closed at all times.
 Persons not immune to varicella should not enter the room. Alternate caregivers should be assigned to
the patient.
Recommendations for Airborne Contact Precautions include:
 If susceptible persons must enter the room of a patient with varicella, an isolation mask should be worn
but this may not be completely protective.
 Put on a gown, and gloves before entering the patient’s room. As part of Standard Precautions, a mask
and eye protection are required for close contact (within 3 feet) with a coughing patient.
 Remove gown, and gloves just inside the door to the patient’s room and discard within the room.
 Wash hands thoroughly after gown and glove removal.
Airborne Precautions
 Airborne Respiratory Precautions are used to prevent the spread of TB (Mycobacterium
Tuberculosis),Measles, Severe Acute Respiratory Syndrome (SARS), Herpes Zoster (Shingles), and
Varicella (Chickenpox).
47
 TA SLIDES BOOK SERIES
 Airborne transmission occurs by dissemination of airborne droplet nuclei, tiny particles containing
microorganisms that remain suspended in the air for long periods.
 These microorganisms are generated from the infected person, generally by coughing, sneezing, and
talking, or by procedures which generate secretions (suctioning, bronchoscopy, etc.) and may be inhaled by
a susceptible host.
Negative pressure, single room is required:
 The door to the room must remain closed at all times.
 Wash hands thoroughly or use alcohol hand gel before
entering and after exiting the room.
 Use a powered air-purifying respirator (PAPR) or N-95
respirator when in the room.
 Visitors will be restricted to parents or 2 designated
primary caregivers.
 Parents / primary caregivers will be screened for
evidence of pulmonary tuberculosis and will be isolated to
the room with the patient until the evaluation is completed.
 Patients may not leave the isolation room except for
essential tests or procedures that cannot be performed in the
room.
 If possible, the patient wears an isolation mask when out of the room.
Enteric Precautions
 Enteric precautions designed to prevent infections that are transmitted by direct or indirect contact with
fecal material, such as Salmonella gastroenteritis.
 Used to control diseases that can be transmitted through direct or indirect oral contact with infected
faces, or contaminated articles.
 E.g. Hepatitis A, Dysentery.
Recommendations for Enteric Precautions
 Place patient in a single private room, put on a gown and gloves before entering the patient’s room.
 Remove gown and gloves just inside the door to the patient’s room and discard within the room.
 Wash hands thoroughly after glove and gown removed.
 Gowns should only be worn when handling object contaminated with faces.
Infections which requires Enteric Precautions:
 Diarrhea Infections or of unknown origin:
• Clostridium Difficile. • Norovirus • Rotavirus • Hepatitis A
Strict Precautions
 Used to prevent the transmission of all highly communicable disease that are spread by both contact and
airborne routes of transmission.
 Strict Isolation Designed for highly contagious infections that are spread by both airborne droplet nuclei
and contact transmission.
 E.g. chickenpox, Rabies
Recommendations for Strict Precautions
 Place patient in a single private room, put on a gown, mask, and gloves before entering the patient’s
room.
 Remove gown and gloves just inside the door to the patient’s room and discard within the room.
 Wash hands thoroughly.
 Private room with negative airflow.
Infections which requires Strict Precautions:
 Infection tuberculosis  Cholera  Varicella
 Disseminated herpes zoster  Diphtheria  yellow fever
 Severe acute respiratory syndromes.
Protective Precautions
 Protective isolation is used for severely immunocompromised patients in order to prevent contamination
and/or infection with microorganisms (bacteria and viruses). For example, protective isolation is used for
patients with burns or leukemia.

48
 TA SLIDES BOOK SERIES
 Used to prevent contact between potentially pathogenic microorganism and uninfected persons who are
seriously impaired resistance.
 Patients with certain diseases, such as leukemia, who are on certain therapeutic regimens are
significantly more susceptible to infections.
Recommendations for Protective Precautions
 Gloves  Mask and protective eye gears.
 Gown  Contaminated needle should not be recapped
 Used puncture resistance containers for used needles and other sharp items.
Infections which requires protective Precautions:
 Immunocompromised patients.  leukemia  Infected burns
Wound and skin Precautions
 Wound and skin precautions are used to prevent the spread of microorganisms found in infected wounds
(including burns and open sores) and contact with wounds and heavily contaminated articles.
 Conditions requiring these precautions include infected burns, infected wounds.
 Diseases that may require wound and skin precautions include herpes, impetigo, and ringworm.
Recommendations for wound and skin Precautions
 Gloves
 Mask and protective eye gears.
 Gown
 Contaminated needle should not be recapped
 Used puncture resistance containers for used needles and other sharp items.
Infections which requires wound and skin Precautions:
 Bacteria cause cellulitis, impetigo, and staphylococcal infections.
 Herpes simplex virus.  Chickenpox
 Fungi cause athletes foot and yeast infections.  Parasite cause body lice, and scabies.
 Skin Diphtheria-(Corynebacterium diphtheria).
Blood Precautions
 Blood precautions are used to prevent acquisition of infection by patients and personnel from contact
with blood or items contaminated with blood. Examples of diseases that require blood precautions .
 Infections which requires blood Precautions are HBV and HIV/AIDS.
Recommendations for Blood Precautions
 Gloves  Mask and protective eye gears.
 Gown  Contaminated needle should not be recapped
 Used puncture resistance containers for used needles and other sharp items.
Discharge Precautions
 Secretion precautions- These precautions are used to prevent acquisition of infection by personnel and
patients from direct contact with wounds and secretion-contaminated articles. Some examples of diseases
requiring these precautions are conjunctivitis, gonorrhea, and syphilis.
 Secretion precautions-oral. These precautions are used to prevent acquisition of infection by personnel
from direct contact with oral secretions. Some examples of diseases requiring these precautions are herpes
areolas and scarlet fever.
 Excretion precautions. These precautions are used to prevent acquisition of infection by personnel and
patients from direct contact with fecal excretions. Some examples of diseases requiring these precautions are
staphylococcal food poisoning.
Recommendations for Discharge Precautions
 Mask and protective eye gears.  Gown
 Contaminated needle should not be recapped  Gloves
 Used puncture resistance containers for used needles and other sharp items.
Breaking the Chain of Infection
 Patient’s scenario:
 A patient assigned for morning care has an open wound on her left lower leg. The wound is draining
and when last cultured, the micro organism MRSA was identified.
 In preparation of bed making, hands of the nurse were washed, clean linen and a bag for soiled linen
were gathered from the linen room and placed on the patient’s clean bed side stand.

49
 TA SLIDES BOOK SERIES
Breaking the Chain of Infection
 Infection Agent: MRSA
 Reservoir: (Patient’s infected wound)
 Portal of Exit: (Draining from the open wound)
 Mode of transmission: MRSA commonly transferred on hands of the nurse by indirect contact.
 BREAK IN THE CHAIN:
 Nurse used proper hand washing techniques, worn protective gloves and properly handled the linen.
 Proper hand washing, gloving and handling of linen.
 To remove the soiled linen from the bed, following procedure was
followed:
 Hands washed
 Gloves worn
 Each side of the soiled linen ends folded towards the middle of the bed
 Soiled linen held away from the nurse’s clean uniform
 Soiled linen placed in the linen bag for later discard
 Protective gloves removed.
 Hands washed
POINT TO BE NOTED:

50
 TA SLIDES BOOK SERIES
Normal Flora
Normal flora is the mixture of microorganisms (bacteria and fungi) that are regularly found at any
anatomical site of human body.
Normal flora are also called commensals (i.e. organism that dine together) or microflora.
Under normal condition in a healthy human they are harmless and may even be beneficial.
Normal flora is not present inside the vital organs of the body, because vital organs are sterile e.g. brain,
kidney, heart, urinary bladder.
There is no occurrence of normal flora inside the vital organs.
Symbiotic Relationship
Mutualistic: both organism benefit- “mutually beneficial” e.g. E coli Synthesis vitamin K & B complex
vitamins In return we provide a warm, moist nutrient rich environment for E. coli.
Commensalistic: one organism benefits, the other is neither helped nor harmed.
Opportunistic: under normal condition ,microbes does not cause disease, but if condition become
conducive, it can cause disease. E. coli normally in our digestive tract where it cause no problems, but if it
gets into the urinary tract it become pathogenic.

Origin of Normal Flora

A fetus, is sterile when born ,then new born start having the normal flora when passing through mothers
vaginal tract and then go to environment ( air, food,etc)
With in few hours, oral and nasopharyngeal flora of neonates establishes
With in one day, resident flora of lower intestinal tract establishes.
Factors Influencing the Normal Flora
Local Environment Diet Age
Sex Immune Condition Use of antibiotics
Types Of Normal Flora
Resident Normal flora: are microorganisms that are always present on or in a person. Resident flora are
life-long members present at certain anatomical sites.
Consists of relatively fixed types of microorganism regularly found in a given area at a given site, if
disturbed, it re-establishes itself.
When the number of resident flora is greatly reduced, opportunistic microbes can easily infection in the
area.
e.g. candida albican that cause candidiasis.
Transient Normal flora: are microorganisms that colonize people for hours to weeks but do not establish
themselves permanently. Transient microbes are unable to colonize the body for longer periods.
Microbes that live in or on your body for a period of time then move on or die off.
51
PAGE NO
 TA SLIDES BOOK SERIES
These are either pathogenic or non-pathogenic.
It varies from time to time/environment.
Transient flora inhibit the skin or mucus membrane for hours (or) days (or) weeks.
Does not re-establishes by itself.
Differences between Resident and Transient Normal Flora
Resident Organisms Transient Organisms
In deeper layer of skin In superficial layer of skin
Permanent flora Temporary flora
If disturbed reestablish themselves Usually does not reestablish themselves
Not removed by routine hand wash Easily removed by routine hand wash
Usually not associated with transmission Usually associated with transmission
of infection of infection
Beneficial Role of Normal Flora
Provide a first line of defense against microbial pathogens through “bacterial interference”.
Assist in digestion: They produce vitamin B and vitamin K in intestine.
Play a role in toxin degradation.
The oral flora contribute to immunity by inducing low levels of circulating and secretory antibodies that
may cross react with pathogens.
Protect the host from colonization with pathogenic microbes
The microbiota affect carcinogenesis in three broad ways:
(i) Altering the balance of tumor cell proliferation and death.
(ii) Regulating immune system function .
(iii) Influencing metabolism of host-produced factors, foods and pharmaceuticals such as : Lactobacillus
and Bifidobacteria, are known to prevent tumor formation.
Harmful Effects of Normal Flora
When host immune system decreased then become opportunistic pathogen and cause disease.
Some may cause diagnose confusion.
Some penicillinase producing microorganism can aggregate and develop antibiotic resistance (Drug
resistant).
Distribution and Occurrence of the Normal Flora
Normal flora of the skin Normal flora of the eye
Normal flora of the digestive tract Normal flora of the Gastro Intestinal Tract
Normal flora of the respiratory tract Normal flora of the urinary tract
Normal flora of the genital tract
Normal Flora of the Skin
The resident microflora of the skin primarily bacteria and fungi.
Resident bacteria of the skin can be in any layer of the skin.
Skin can acquire any transient bacteria from the environment but it either get washed off or die because
the skin is dry, has acidic pH and produce sweat and oil.
Resident flora of the skin (staphylococcus-epidermis, staphylococcus aureus, streptococcus, diptheriods,
propionibacterium acne, yeast-candida).
Transient flora of the skin ( micrococcus, bacteriods ).

52
PAGE NO
 TA SLIDES BOOK SERIES
Risk Factors of the skin Microbiome

Normal Flora of the eye

Low number present due to: High moisture
Blinking of the eye mechanically remove the bacteria Lachrymal secretions include lysozyme
The conjunctiva of the eye has primarily S.epidermidis, S.aureus, C. diphtheroid, and S.pneumoniae.
Neisseriae and gram-negative bacilli resembling haemophili (Moraxella species) are also frequently
present.
Normal Flora of the Digestive Tract
Microflora of oral cavity:
They have both aerobic and anaerobic bacteria
The most common ones are: C.diptheroids, S.aureus,
Epidermis.
Also, yeast, can be living in mouth.
Mouth/Oral cavity: lactobacilli, staphylococcus,
streptococcus, bacteroides, corynebacterium, etc.
Teeth: streptococcus mutants, hemolytic streptococci,
fusobacterium, etc.
Normal Flora of the Gastro Intestinal Tract
Stomach: e.g.: acid tolerant streptococci, staphylococci, lactobacilli, candida albican, and helicobacter.
Small Intestine:
Duodenum: Gram positive cocci and rods are more.
Jejunum: Gram positive, Gram negative bacterium can be seen, Lactobacilli diptheroids, enterococcus,
faecalis, yeast- candida, albicans.
Ileum: pH-alkaline, anaerobic gram positive bacteria are more mainly entero bacterial, shigellosis.
Large Intestine:
Largest microorganism in the body anaerobic gram positive, non sporing bacteria.
Bacteriods, lactobacillus, E.coli, proteus, yeast, candida, protozoans, entamoeba hystolitica, trichmonas.

53
PAGE NO
 TA SLIDES BOOK SERIES

Normal Flora of the Respiratory Tract

Upper respiratory Tract: Nasal passage,
Esophagus
The organism which are involved are
staphylococcus, streptococcus, diptheriod,
bacteroides, micrococci, and haemophilus.
Lower respiratory Tract: trachea, bronchi,
lungs
It is mostly sterile, because the mucous
membrane of the lungs remove any
microbes.
Normal Flora of the Genitourinary Tract
Normal flora of Urinary tract:
Urethra and urinary bladder are sterile.
Non pathogenic gram positive cocci and
gram negative enterobacteriaceae members
may be extremely found as normal flora.
Mycobacterium smegmatis is a commonly
present in both male and female genital tract.
Male Genital tract:
Lactobacillus bacteroids, staphylococci, and
corynebacterium.
Female Genital tract:
The adult female genital tract has complex
normal flora from puberty to menopause.
The pH is acidic 4.2-4.6, acidophilic colonizing
bacteria e.g. Lactobacilli known as doderline
bacilli, staphylococcus epidermis, streptococci,
peptostreptococci, clostridium species,
54
PAGE NO
 TA SLIDES BOOK SERIES
diptheriods,candida.
Nosocomial Infections
Nosocomial also Called Hospital acquired infection.
DEFINITION (WHO): - An infection acquired in hospital by a patient who was admitted for a reason other
than that infection.
An infection occurring in a patient in a hospital or other health care facility in whom the infection was
not present or incubating at the time of admission.
This includes infections acquired in the hospital but appearing after discharge & also occupational
infections among staff of the facility.
Around 9% of all hospital patients develop hospital infection (nosocomial).
Common Nosocomial Infections
Urinary tract infections (UTI). Surgical wound infections
(SWI).
Lower respiratory infections. Traumatic wounds and
burns infections.
Primary bacteremia. Gastrointestinal tract.
MODE OF TRANSMISSION:
Contact/hand borne (most common) Aerial route or air borne
Oral route Parenteral route
Vector borne
Sources of Nosocomial infection
Lower defenses Antibiotic treatment
Contaminated hospital Environment, e.g. fluid, blood, food, air.
Patients flora (cutaneous, GIT flora, Respiratory flora)
Medical or surgical procedures used: catheters medical equipment Medical personals.
Measures to prevent and control Nosocomial infections
Treat all patients as a potential biohazard.
Isolation: Designed to prevent transmission of microorganisms by common routes in hospitals.
Sterilization: of all reusable equipment's such as ventilator, humidifier and any device that come in
contact with the respiratory tract.
Adopt universal routine safe infection control practices to protect patients, self and colleagues from
infection.

55
PAGE NO
 TA SLIDES BOOK SERIES
Standard Precautions
I. Hand hygiene.
II. Use of personal protective equipment (e.g., gloves, gowns, masks).
III. Safe handling of potentially contaminated equipment/surfaces/sharp disposal in the patient
environment.
IV. Respiratory hygiene. V. Safe injection practices. VI. Use of mask.

Universal precautions
Hand washing Personal protective equipment [PPE]
Preventing/managing sharps injuries Aseptic technique
Isolation Staff health
Linen handling and disposal Waste disposal
Spillages of body fluids Environmental cleaning
Risk management/assessment
Personal protective Equipment's
 PPE when contamination or splashing with blood or body fluids is anticipated:
Disposable gloves Plastic aprons
Face masks Safety glasses, goggles, visors
Head protection Foot protection
Fluid repellent gowns
REMEMBER:
EVERYTHING YOU TOUCH HAS BEEN TOUCHED BY SOMEONE ELSE.
Hand hygiene is the simplest, most effective measure for preventing hospital-acquired infections.

56
PAGE NO
 TA SLIDES BOOKSERIES

Unite 06 PART 2 DISEASE

 In this unit we will discuss the following Diseases:
 Tetanus  Tuberculosis  Typhoid
 Cholera  Diphtheria  Pertusis
 Mumps  Measles  Polio
 Influenza  Ascariasis  Teaniasis
 Dermatomycosis
Tetanus
 Tetanus is an acute, often fatal, disease caused by an exotoxin produced by the bacterium Clostridium
tetani.
 It is characterized by generalized rigidity and convulsive spasms of skeletal muscles.
 The muscle stiffness usually begins in the jaw (lockjaw) and neck and then becomes generalized.
 Tetanus infection can be life threating without treatment. Approximately 10 to 20 percent of tetanus
infection are fatal, according to the centers of disease control and prevention(CDC).
 Nicolaier isolated the bacterium from soil in 1884.
 In 1889, Kitasato-Shibasab isolated the organism from human, show that it produced disease when
injecting into animals.
 In 1897 Edmond Nocard demonstrated protective effect of positively
transferred antitoxin during word war I.
 However first effective tetanus vaccine was develop by Gaston Romon in 1920.
 Tetanus toxoid developed in 1924 and widely used during World War II.
 It is important to note that tetanus can be prevented by getting vaccinated and seeking medical attention
immediately if there is an injury or wound that may be contaminated with tetanus spores.
 Combine 1948 name(DTP).
Reservoir of infection
 The natural habitat is found in the soil and dust. The bacilli are also found in the intestine of animals
such as cattle, hourse, goats, sheep, and excreted with their faces. The spore survive for year in soil.
Mode of Transmission
 By contact of Spore with wound  wound contamination
 Infected formites  Soiled or infected dressing on wound
 Umbilical cord sepsis  Surgical incision
 Middle ear infection
 Incubation Period: 4 to 21 days  Rout of Entry: Wound
 Occurrence: Worldwide
 Prevention & Control: Active immunization, public education.
 Treatment:
 Antitoxin  TIG ( TETANUS IMMUNOGLOBULINE)
 Debridement of wound  Benzylepenicillin 600 mg 6 hourly I/V
 Diazepam
Clinical Features
 Risus sardonicus (devil smile)- due to facial muscle
spasm
 Lock jaw  Difficulty in swallowing
 Restlessness  Irritability
 Headache  Fever
 Tachycardia
Pathophysiology
 Tetanus toxin is a potent neurotoxin produced by the bacterium Clostridium tetani. The mechanism of
action of tetanus toxin involves binding to nerve cells and blocking the release of neurotransmitters, which
are chemical messengers that transmit signals between nerve cells and muscles.
 The tetanus toxin is composed of two chains, the light chain and the heavy chain. The heavy chain is
responsible for binding to nerve cells, while the light chain contains the enzymatic activity that causes the
toxic effects.
 After binding to nerve cells, the tetanus toxin is taken up into the cell and the light chain is released.
The light chain then cleaves a protein called synaptobrevin, which is required for the release of
57
PAGE NO
 TA SLIDES BOOKSERIES

neurotransmitters from nerve cells. This prevents the release of the neurotransmitter called GABA
(gamma-amino butyric acid), which is responsible for inhibiting muscle contraction.
 Without the inhibition provided by GABA, muscles become hyperactive and uncontrollable, leading to
the characteristic muscle stiffness and spasms associated with tetanus.
 The severity of tetanus depends on the amount of toxin produced and the host's immune response. In
severe cases, the toxin can spread throughout the body and affect multiple organ systems, leading to
respiratory failure, cardiac arrest, and death.
Nursing Management
 Isolation of patient
 Keep patient in a darkened room to prevent photophobia, over stimulation and spasms.
 Ensure a quiet environment to prevent spasms
 Respiratory support  Pain Management  Employ barrier nursing techniques
What is Tuberculosis
 TB is an infectious disease caused by the bacillus Mycobacterium tuberculosis.
 The bacteria usually attack the lungs, but they can also damage other part of the body .
 TB spread through the air when a person with TB of the lung or throat coughs.
 Robert Koch discovered mycobacterium tuberculosis in 1885.
 In 2016 worldwide 10.6 million people became sick with TB and 1.7 million TB-related deaths.
 Over 10 million in the US are infected and they have a lifelong risk of developing TB.
 Without treatment ,approximately 5-10% of patients with latent TB will progress to active TB diseases
at some point in their lifetime.
 Isoniazid can reduce the incidence of active TB about 90%.
Types
 Types:
1. Pulmonary TB - typically affects the lungs
2. Extra-pulmonary TB - can affect other sites
1. Pulmonary TB - typically affects the lungs.
 Persistent cough for 3 weeks or more
 Productive cough with or without blood stained sputum
 Shortness of breath and chest pain
 Intermittent fever, night sweats, loss of weight, fatigue.
2. Extra-pulmonary TB - can affect other sites
 TB of bones and joints:
 Localized pain and /or swelling, pus discharge, muscle weakness, paralysis and stiffness of joints.
 Intestinal TB:
 Loss of weight and appetite  Abdominal pain, diarrhea, constipation
 Mass in abdomen  Ascites (fluid in the abdominal cavity)
 Tuberculosis meningitis:
 Headache  Fever
 Vomiting  Neck stiffness and mental confusion of insidious onset.
Pathophysiology
 Entry: TB bacteria enter the body through the respiratory tract, usually by inhaling contaminated air
droplets from an infected person who is coughing or sneezing.
 Phagocytosis: Once inhaled, the bacteria are engulfed by immune cells called macrophages, which
normally engulf and destroy invading pathogens. However, TB bacteria have the ability to evade destruction
by the macrophages.
 Granuloma formation: The TB bacteria are able to survive and multiply inside the macrophages, which
then migrate to the lymph nodes and form granulomas, which are small nodules composed of immune cells
that surround the bacteria to prevent them from spreading.
 Latency: In some cases, the immune system is able to control the infection, and the bacteria become
dormant inside the granulomas, a condition known as latent TB infection.
 Reactivation: In some cases, the bacteria can break out of the granulomas and start multiplying, leading
to the development of active TB disease, which can cause symptoms such as cough, fever, weight loss, and
night sweats.

58
PAGE NO
 TA SLIDES BOOKSERIES

 Spread: Active TB disease can spread to other parts of the body, including the lungs, bones, and brain.
The bacteria can also be spread to other people through the air when an infected person coughs or sneezes.
Evaluation for TB
 Medical history  Physical examination
 Mantoux tuberculin skin test  Chest x-ray
 Bacteriologic exam (smear and culture)
Mode Of Transmission
 Tuberculosis is a droplet (Air born) infection; when people with lung TB cough, sneeze or spit, they
propel the TB germs into the air.
 Through quick, casual contact.
 By sharing utensils or food
 By sharing cigarettes or drinking containers
 By shaking hands
 Using public telephones
 Nose and throat (by inhalation) are commonest point of entry.
 Mouth (by ingestion) is another point of entry for
Mycobacterium tuberculosis.
 Incubation Period: 4-12 weeks  Occurrence: Pandemic
Source Of Infection
 Most common source of infection is a case of tuberculosis whose sputum is positive for Mycobacterium
tuberculosis, who has received no treatment or incomplete/irregular treatment.
 Other sources of infection are excreta of such patient, milk obtained from cows suffering from TB and
material from slaughtered tuberculosis animal.
Clinical manifestations of TB

Drug-resistant TB
 Drug-resistant TB is caused by:
 Inconsistent or partial treatment, when patients do not take all their medicines regularly for the required
period because they start to feel better.
 Doctors and health workers prescribe the wrong treatment regimens.
 The drug supply is unreliable.
Multidrug-Resistant TB (MDR-TB)
 Multidrug-resistant tuberculosis (MDR-TB) is a form of TB caused by bacteria that do not respond to,
at least, Isoniazid and Rifampicin, the two most powerful, first-line (or standard) anti-TB drugs.
 The primary cause of MDR-TB is inappropriate treatment.
 MDR-TB is treatable and curable by using second-line drugs.
 The extensive chemotherapy required (up to two years of treatment) is more costly and can produce
severe adverse drug reactions in patients.
 In some cases more severe drug resistance can develop.
Extensively drug -resistant TB (MDR-TB)
 Extensively drug-resistant TB (XDR-TB), is a form of multi-drug resistant tuberculosis that responds
to even fewer available medicines, including the most effective second-line anti-TB drugs.
 About 450, 000 people developed MDR-TB in the world in 2012.
 It is estimated that about 9.6% of MDR-TB cases had XDR-TB.

59
PAGE NO
 TA SLIDES BOOKSERIES

Prevention and control

 Chemotherapy of cases
 Chemoprophylaxis for contacts
 Immunization of infants with BCG ( Bacillus Calmette Guerine)
 Educate patients with TB about the mode of disease transmission and how to dispose their sputum and
cover their mouth while coughing, sneezing, etc.
 Vaccination - BCG Public education, adequate diagnostic and curative facilities, Isolation,
Environmental sanitation.
 Public health education about the modes of disease transmission and methods of control:
 Improved standard of living  Adequate nutrition
 Environmental sanitation  Personal hygiene; etc.
 Active case finding and treatment  Avoid public places
 Limit visitors
Nursing care
 Educate the patient how and when to take the prescribed medication.
 Tell the patient not to stop the medication unless he/she is told to do so.
 Tell the patient to come to the health institution if he/she develops drug side effects.
 Advice the patient on the importance of taking adequate and balanced diet and to eat what is available at
home.
 Involve the family in teaching on medication.
 Tell the patient that Rifampicin colors the urine orange.
Remember
“A decision to test is a decision to treat.”
Typhoid Fever
 A systemic bacterial infectious disease characterized by high prolonged fever, malaise, caused by the
bacterium salmonella tophi.
 Typhoid is bacterial infection that can lead to a high fever, diarrhea and vomiting, Fatigue, Abdominal
pain.
 It can be fatal.
 Causative Organism: Salmonella Typhi
 Salmonella typhi is a gram negative rod, shaped flagellated bacterium whose only reservoir is the
human body.
 According to WHO, an estimated 11-20 million people get sick from typhoid and between 128,000 and
161,000 people die from it every year.
Reservoir
 The mode of transmission of typhoid fever is through the consumption of food or water that has been
contaminated with the feces of an infected person. The bacteria then enter the body through the digestive
system and invade the bloodstream, leading to the development of symptoms.
 Man is also known reservoir of infection-( carriers)
Pathophysiology
 Incubation period: After the bacteria enter the body, there is an incubation period of 1-3 weeks before
symptoms start to develop.
 Bacterial invasion: The bacteria invade the lining of the small intestine, where they multiply and spread
into the bloodstream.
 Septicemia: The bacteria multiply rapidly in the bloodstream, causing septicemia (blood poisoning).
This leads to the development of symptoms such as fever, chills, and weakness.
 Spread to other organs: The bacteria can spread to other organs, including the liver, spleen, and bone
marrow, leading to the development of symptoms such as abdominal pain, diarrhea, and headache.
 Recovery: Most people with typhoid fever recover within a few weeks with appropriate treatment, but
some may become chronic carriers of the bacteria, meaning they continue to harbor the bacteria in their
intestine without showing any symptoms. These carriers can continue to shed the bacteria in their feces and
and infect others.
 Incubation period : 1-3 weeks
 Period of communicability: A case is infectious as long as the bacilli appear in stool .
 Occurrence: Pandemic (Worldwide) Salmonellae are gram -ve rods, facultatively aerobic
60
PAGE NO
 TA SLIDES BOOKSERIES

Predisposing Factors
 Age group: At any age but mainly in children and young adults.
 Occupation: Working environment
 Socio-economic factors: Inadequate sanitation facilities and unsafe water supplies.
 Nutritional status: Mal-nutrition
Clinical manifestations
 First week Fever, headache, myalgia, relative bradycardia, constipation, Leucopoenia—White cell
count is less than 4000/mm
 Second week Rose spots on chest and trunk, splenomegaly, abdominal distention and tenderness,
diarrhea
Rose spots
 Diagnosis:
• Clinical presentation • ƒWidal test (but not as much reliable)
• Typhidot (IgM-recent infection , IgG-remote infection) • Blood culture
 Treatment:
• ciprofloxacin 500 mg B.D or ceftriaxone for 14 days. • Chronic carriers should be
treated for 4 weeks
Prevention and Control
 MEASURES DIRECTED TO RESERVOIR:
a) Case detection and treatment b) Isolation from handling food etc.
c) Education on hand washing c) Disinfection of stools and urine
d) Detection & treatment of carriers
 MEASURES AT ROUTES OF TRANSMISSION:
a) Water & Food sanitation b) Excreta disposal
 MEASURES FOR SUSCEPTIBLES:
a) Immunization b) Health education
Cholera
 Cholera is an acute diarrheal infection caused by ingestion
of food or water contaminated with the bacterium Vibrio
cholera. Cholera remains a global threat to public health and an
indicator of inequity and lack of social development.
 Definition: A severe acute gastrointestinal infection which
is characterized by rice watery stool.
 Cholera is a diarrheal disease caused by the bacterium
Vibrio cholera.
 10-20% of cholera patients develop severe watery diarrhea
with vomiting.
 Outbreaks can occur where water supply, sanitation, food
safety and hygiene are inadequate.
 Greater risks occur in over-populated communities and refugee camps, with Poor sanitation, Unsafe
drinking water, Increased person to-person transmission.
 Because incubation period is very short, the number of cases can rise extremely rapidly.
 Treatment is straight forward (basically rehydration), and if applied correctly and promptly, case fatality
rate is <1%.
 In untreated cases, case fatality rate may reach 30-50%.
Transmission of Cholera
 Transmitted by the fecal-oral route.
 Contaminated food (especially seafood) is a more common cause of cholera in developed countries,
whereas contaminated water is more common in developing countries.
 88% of all diarrheal disease in the world can be attributed to unsafe water, sanitation and hygiene.
Pathophysiology
 Infection: Cholera is transmitted through the ingestion of contaminated food or water that contains the
bacterium. Once inside the body, the bacteria begin to multiply in the small intestine.

61
PAGE NO
 TA SLIDES BOOKSERIES

 Toxin production: The bacteria produce a toxin called cholera toxin, which affects the cells lining the
small intestine. Cholera toxin binds to the surface of the intestinal cells, causing them to release large
amounts of water and electrolytes.
 Fluid loss: The release of water and electrolytes leads to the characteristic symptoms of cholera,
including severe diarrhea and dehydration. The diarrhea is often described as "rice water" because it is
watery and contains flecks of mucus.
 Spread: The bacteria can be shed in the stool of infected individuals and can contaminate water sources,
leading to the spread of the infection to others.
 Treatment: Cholera is treated with rehydration therapy, which involves replacing the fluids and
electrolytes lost due to diarrhea and dehydration. Antibiotics can also be used to shorten the duration of
diarrhea and reduce the spread of the bacteria.
 S/S: Fever, vomiting, rice watery stool, malaise, dehydration
 Causative organism: Vibrio cholera A comma shaped Gram -ve bacillus
 Incubation Period: Hours to 5 days
 Source and reservoir of Infection: Faeces and vomitus of infected and carrier person
 Rout of Entry: Oro - faecal
 Mode of transmission: Ingestion of contaminated food, water & 5 Fs ( fluids, fingers, flies, foods, and
fields.)
 Occurrence: Endemic in Asian countries where sanitation is poor.
 Prevention & Control: Active immunization, pure food and water supply, sanitary disposal of human
excreta, fly control, public education.
 Management: Fluid and electrolyte replacement, Tetracycline, cotrimaxazole.
Control and Prevention
Food Protection & Safe Food Social gatherings, market places.
Key Messages:
 Cook foods thoroughly  Store foods carefully
 Reheat foods thoroughly  Avoid contacts between foods that is raw and uncooked
 Wash hands repeatedly during food preparation
Safe Water
 Protection of water sources
 Treatment of water sources:
o Chlorination, filtration
 Safe Water collection and storage practices:
o Narrow-mouthed vessels
o Chlorination
 Household Water Treatment:
o Boiling
o Chlorination
Diphtheria
 Definition: An acute bacterial infection of the upper respiratory
tract (throat, nose & larynx) characterized by the formation of grayish
membrane in throat.
 S/S: Inflammation of throat, pseudomembrane, dry cough
 Causative organism: Corynebacterium diphtheria or Klebs
Loeffler bacillus. It is Gram +ve bacillus. It releases exotoxin.
 Incubation Period: 2 to 5 days
 Reservoir & Source : Reservoir is infected man and source is
nasopharyngeal secretion.
 Rout of Entry: Inhalation
 Mode of transmission: by contact with patient or carrier and infected fomites.
 Occurrence: Worldwide
 Prevention & Control: Active immunization, Isolation, concurrent and
terminal disinfection of infected fomites.
 Treatment: Benzylpenicillin 600 mg 6 hourly I/V for 7 days.

62
PAGE NO
 TA SLIDES BOOKSERIES

Pathophysiology
 Infection: Diphtheria is transmitted through respiratory droplets or contact with the skin lesions of
infected individuals. The bacteria then multiply in the respiratory tract or on the skin.
 Toxin production: The bacteria produce a toxin called diphtheria toxin, which affects the cells in the
body. The toxin is carried by the bloodstream to the tissues and organs.
 Cell damage: The toxin binds to the surface of human cells and is taken up by the cells. Once inside the
cells, the toxin inactivates a protein called elongation factor 2 (EF-2), which is needed for protein synthesis.
This leads to the death of the cells in the affected tissues
 Pseudomembrane formation: In severe cases of diphtheria, the dead cells, fibrin, and other materials can
accumulate in the affected tissues and form a thick, grayish-white membrane. This membrane can block the
airways and cause breathing difficulties.
 Spread: The bacteria can be shed in the respiratory secretions or skin lesions of infected individuals and
can be spread to others through close contact.
 Treatment: Diphtheria is treated with antitoxin, which neutralizes the diphtheria toxin and prevents
further damage to the cells. Antibiotics can also be used to kill the bacteria and prevent further spread of the
infection.
Pertussis (History)
 The earliest clear account of whooping cough was described in 1640 by Baillow, an epidemiologist
 The name „pertussis‟ means “violent cough”, and was first used to describe the disease in 1679.
 In China, the disease is known as “Hundred Day Cough”
PERTUSSIS (WHOOPING COUGH)
Whooping cough: whooping sound made when gasping for air after a fit of coughing Cough of 100
days
Introduction
 A highly contagious acute bacterial infection caused by the
bacilli Bordetella pertussis
 Currently worldwide prevalence is diminished due to active
immunization
 However it remains a public health problem among older
children and adults
 It continues to be an important respiratory disease afflicting
unvaccinated infants and previously vaccinated children and adults (waning immunity).
Definition
 An acute bacterial infection of respiratory tract involving trachea, bronchi and bronchioles characterized
by a typical cough.
 S/S: Crowing like Whooping sound cough, paroxysmal cough, tenacious mucous, vomiting, catarrhal
S/S
 Causative organism: Bordetella pertussis It is Gram -ve bacillus.
 Incubation Period: 7 to 21 days
 Reservoir & Source: Reservoir is infected man and source is branchopharyngeal secretion.
 Rout of Entry: Inhalation
 Mode of transmission: By droplet through direct contact with infected patient and indirect through
infected fomites.
 Occurrence: Worldwide. Children below five years are more susceptible.
 Prevention & Control: Active immunization, Isolation, concurrent and terminal disinfection of
infected fomites
 Treatment: Erythromycin 250 mg 6 hourly Antitussive drug
Pathophysiology
 Infection: Whooping cough is transmitted through respiratory droplets when an infected person coughs
or sneezes. The bacteria then attach to the cilia (hair-like structures) that line the respiratory tract.
 Toxin production: The bacteria produce a toxin called pertussis toxin, which affects the cells in the
respiratory tract. The toxin is carried by the bloodstream to other tissues and organs in the body.
 Inflammation and coughing: The bacteria and their toxins cause

63
PAGE NO
 TA SLIDES BOOKSERIES

inflammation in the respiratory tract, leading to a persistent cough that can last for several weeks. The
coughing fits can be severe and may be accompanied by a "whooping" sound when the person tries to take a
breath.
 Complications: In severe cases of whooping cough, the persistent coughing fits can lead to
complications such as pneumonia, seizures, and even brain damage.
 Spread: The bacteria can be shed in the respiratory secretions of infected individuals and can be spread
to others through close contact.
 Treatment: Whooping cough is treated with antibiotics, which can kill the bacteria and prevent further
spread of the infection. Vaccines are also available for the prevention of whooping cough.
Treatment
 Avoidance of irritants, smoke, noise and other cough promoting factors
 Antibiotics: effective only if started early in the course of illness. Erythromycin (40-50 mg/kg/day 6
hrly orally for 2 weeks or Azithromycin 10 mg/kg for 5 days in children<6 months and for children>6
months 10 mg/kg on day 1, followed by 5mg/kg from day2-5 or Clarithromycin 15 mg/kg 12 hrly for 7 days
 Supplemental oxygen, hydration, cough mixtures and bronchodilators (in individual cases)
Prevention
 All household contacts should be given erythromycin for 2 weeks
 Children <7 years of age not completed the four primary dose should complete the same at the earliest
 Children <7 years of age completed primary vaccination but not received the booster in the last 3 years
have to be given a single booster dose
 VACCINE
Mumps
 Definition
• An acute viral infection primarily of salivary glands characterized by
fever and swelling of usually parotid glands.
 S/S: Malaise, anorexia, fever, sore throat, parotitis and swelling,
tenderness at angle jaw
 Causative organism: Mumps Virus
 Incubation Period: 12 to 26 days
 Reservoir and Source of Infection: Infected person and source is infected saliva.
 Rout of Entry: Respiratory tract
 Mode of transmission: direct through droplet and indirect via infected fomites.
 Occurrence: World wide especially younger people are more affected
 Prevention & Control: Active immunization-MMR (Measles, Mumps, Rubella) vaccine pure food and
water supply, public education.
Measles
 Definition:
• An acute highly communicable viral disease characterized
by abrupt onset catarrhal signs symptoms followed by fever
 S/S: Catarrhal signs symptoms (running nose,
lacremation), fever, cough, photophobia, Koplics spot , and
On day 3-4 maculo-papular rashes
 Causative organism: Measles virus
 Incubation Period: 10 to 14 days
 Reservoir & Source : Infected person and droplets.
 Rout of Entry: Respiratory tract
 Mode of transmission: By direct contact with infected person and indirect through infected fomites.
 Occurrence: World wide and Common in children Complications: stomatitis, keratitis, Enteritis,
pneumonia, otitis media.
 Prevention & Control: Active immunization-MMR vaccine. Isolation, concurrent and terminal
disinfection of infected fomites.
Influenza
 Definition: • An acute highly communicable viral disease of respiratory tract.
 S/S: chills, fever, headache, and general muscular aches
 Causative organism: Influenza virus (RNA virus)
64
PAGE NO
 TA SLIDES BOOKSERIES

 Incubation Period: 24 to 72 hours

 Reservoir & Source : Reservoir is infected man and source is nosopharyngeal secretion.
 Rout of Entry: Respiratory tract

 Mode of transmission: By droplet through direct contact with infected person and indirect through
infected fomites.
 Occurrence: Worldwide, Children younger are more common
 Prevention & Control: Active immunization, Isolation, concurrent and terminal disinfection ofinfected
fomites.
Poliomyelitis
 Definition: • An acute viral disease involving the nerve cells of spinal cord and brain stem.

 S/S: Fever, headache, stiffness of neck and spine, weakness and flaccid paralysis.
 Causative organism: Polio virus
 Incubation Period: 7-21 days
 Source: Oro-pharyngeal secretion and faeces of infected person.
 Rout of Entry: Oro - faecal
 Mode of transmission : By contact with oro-pharyngeal secretion or faeces of infected person. It
reaches nervous tissues via lymphatic tissues and blood.
 Occurrence: Worldwide. Children from 1-12 years are common victims
 Prevention & Control: Active immunization, Isolation, public education.

65
PAGE NO
 TA SLIDES BOOKSERIES

Ascariasis
 Infectious Agent:
Ascariasis is caused by the soil transmitted
helminthes Ascaris lumbricoides and Ascaris
suum. Both are roundworm intestinal nematodes.
Ascaris lumbricoides is found in humans and
dogs, while Ascaris suum is most commonly
found in pigs, but can infect humans via
consumption of contaminated meat.
 Occurrence: Ascaris lumbricoides is the
most prevalent of all human intestinal
nematodes worldwide
 Causative agent:  Ascaris lumbricoides
 Prevalence:  Over 1 billion people infected worldwide (most prevalent of all intestinal nematodes).
 Transmission:
o Transmission is primarily via ingestion of fecal contaminated soil. Eggs are shed in an infected person‟s
feces but do not become infectious until they have incubated in soil for 2-3 weeks. Once they become
infectious they can be transmitted via contaminated water, agriculture products, fingers, or other fomites.
 Incubation Period:
o Eggs must incubate in soil for 2-3 weeks before they become infectious to humans. Once ingested it takes
approximately 8 weeks for the eggs to develop into an egg-laying adult female worm although symptoms
may manifest earlier.
Communicability/Transmission:
 Human to human transmission of Ascaris spp does not occur because part of the worm‟s life cycle must
be completed in soil before becoming infectious. Soil contamination is perpetuated by fecal contamination
from infected humans or dogs for Ascaris lumbricoides and humans (rarely) or pigs for Ascaris suum. An
infected person may shed eggs for as long as they are infected with an egg-laying adult which may be
several years.
Signs & Symptoms
 85% of cases are asymptomatic
 Fever during migratory phase
 Intestinal obstruction due to a bolus of worms
 Inflammation of internal organs including pancreatitis and
appendicitis
In mild or moderate ascariasis, the intestinal infestation can cause:
Vague abdominal pain. Nausea and vomiting. Diarrhea or bloody stools.
If you have a large number of worms in the intestine, you might
have:
 Severe abdominal pain.  Fatigue.  Vomiting.
 Weight loss or malnutrition.  A worm in your vomit or stool
Diagnosis
 Laboratory Criteria:
 Microscopic identification of Ascaris eggs in feces OR
 Microscopic identification of ascrid larvae in sputum or gastric washings OR
 Identification of adult worms passed from the anus, mouth or nose.
Treatment
 Anthelminthic such as albendazole and mebendazole, are the drugs of choice for treatment of Ascaris
infections.
 Infections are generally treated for 1-3 days.
Prevention and Control Measures
 Routine hand washing with soap and warm water.
 Proper disposal of human waste products such as feces is necessary to prevent contamination of soil.
 Avoid areas where human waste contamination of soil or water is likely.
 Proper removal and disposal of pet waste from outdoor areas
 Thoroughly wash fruits and vegetables to remove soil/fertilizer residue.
66
PAGE NO
 TA SLIDES BOOKSERIES

 Thoroughly cook all fruits and vegetables that may have been in contact with soil produced from human
and animal waste.
 Cook all pork products to the appropriate temperature prior to consumption and wash hands thoroughly
before and after handling raw meat.
Taeniasis
 Taeniasis is an infection caused by the tapeworm, a type of parasite.
Parasites are small organisms that attach themselves to other living
things in order to survive. The living things the parasites attach to are
called hosts.
 Parasites can be found in contaminated food and water. If you
consume contaminated food or drinks, you may contract a parasite that
can live and sometimes grow and reproduce inside your body.
 Taeniasis is an intestinal tapeworm infection caused by eating
contaminated beef or pork. It‟s also known by the following names:
– Taenia saginata (beef tapeworm) – Taenia solium (pork tapeworm)
What are the symptoms of Taeniasis?
 Most people who have taeniasis don‟t have any symptoms. If signs and symptoms are present, they may
include:
– pain – unexplained weight loss – blockage of the intestine
– digestive problems
 Some people with taeniasis may also experience irritation in the perianal area, which is the area around
the anus. Worm segments or eggs being expelled in the stool cause this irritation.
 People often become aware that they have a tapeworm when they see worm segments or eggs in their
stool.
 Infections can take between 8 and 14 weeks to develop.
What causes Taeniasis?
 You can develop taeniasis by eating raw or undercooked beef or pork. Contaminated food can contain
tapeworm eggs or larvae that grow in your intestines when eaten.
 Fully cooking beef or pork will destroy the larvae so that they can‟t live in your body.
 The tapeworm can grow up to 12 feet in length. It can live in the intestines for years without being
discovered. Tapeworms have segments along their bodies. Each of these segments can produce eggs. As the
tapeworm matures, these eggs will be passed out of the body in the stool.
 Poor hygiene can also cause the spread of taeniasis. Once tapeworm larvae are in human stool, they can
be spread through contact with the stool. You should wash your hands properly to help prevent the spread of
the infection.
What are the risk factors for Taeniasis?
 Taeniasis is more commonTrusted Source in areas where raw beef or pork is consumed and where
sanitation is poor. These areas may include:
– Eastern Europe and Russia – East Africa – sub-Saharan Africa
– Latin America – parts of Asia, including China, Indonesia, and South Korea
 According to the Centers for Disease Control and Prevention (CDC)Trusted Source, there are probably
fewer than 1,000 new cases in the United States each year. However, people who travel to areas where
taeniasis is more common are at risk of contracting the disease.
 Taeniasis is more likely to develop in people who have weakened immune systems and aren‟t able to
fight off infections. Your immune system can weaken due to:
– AIDS – an organ transplant – Diabetes/chemotherapy
How is Taeniasis diagnosed?
 See your doctor if you see worm segments or eggs in your stool. Your doctor will ask you about your
health history and recent travel outside of the United States. Doctors will often be able to make a diagnosis
of taeniasis based on the symptoms.
 To confirm the diagnosis, your doctor may order blood tests including a complete blood count (CBC).
They may also order a stool exam to see if eggs or worm segments are present.
Treatment
 Taeniasis is typically treated with medications prescribed by your doctor. Medications for the treatment
of taeniasis include praziquantel (Biltricide) and albendazole (Albenza).
67
PAGE NO
 TA SLIDES BOOKSERIES

 Both drugs are antihelmintics, which means that they kill parasitic worms and their eggs. In most cases,
these medications are provided in a single dose. They can take a few weeks to fully clear an infection. The
tapeworm will be excreted as waste.
 Common side effects associated with these medications include dizziness and upset stomach
What complications are associated with Taeniasis?
 In rare cases, serious complications from the infection can occur. Tapeworms may block your intestines.
This may require surgery to correct.
 In other cases, a pork tapeworm may travel to other parts of your body such as the heart, eye, or brain.
This condition is called cysticercosis can cause other health problems such as seizures or infections in the
nervous system.

How can Taeniasis be prevented?

 The most effective way to prevent taeniasis is to cook food thoroughly. This means cooking meat to a
temperature above 140°F (60°F) for five minutes or more. Measure the meat temperature with a cooking
thermometer.
 After cooking meat, allow it to stand for three minutes before cutting it. This can help destroy any
parasites that may be in the meat.
 Proper hand hygiene is also important for preventing the spread of this disease. Always wash your
hands after using the bathroom and teach your children to do the same.
 Also, drink bottled water if you live in or travel to an area where water must be treated
Dermatomycoses
[Ringworm (Dermatophytosis, Tinea, Trichophytosis, Microsporosis, Jock
Itch, Athlete's Foot)]
Ringworm - also known as dermatophytosis, dermatophyte infection, or
tinea is a fungal infection of the skin.
 “Ringworm” is a misnomer, since a fungus, not a worm, causes the
infection. The lesion caused by this infection resembles a worm in the
shape of a ring, which is why it got its name.
 Ringworm is usually specifically used to describe tinea corporis
(ringworm of the body) or tinea capitis (ringworm of the scalp). It‟s
sometimes used to describe tinea infection in other locations, such as tinea
cruris (ringworm of the groin).
 Ringworm infection can affect both humans and animals. The infection
initially appears as discolored,
often scaly patches on affected areas. These patches typically appear red on
lighter skin or brown-gray on darker skin.
 Ringworm may spread from an affected area to other parts of the body, such as the:
– scalp – feet – hands – nails – groin
Recognizing ringworm symptoms
 Symptoms vary depending on where the infection occurs with a skin infection, you may experience the
following:
– itchiness
– itchy or scaly patches that are red, brown, or gray, or raised areas of skin called plaques
– a round, flat patch of itchy skin
– patches that develop blisters or pustules
– patches that resemble a ring with deeper color on the outside
– patches with edges that are defined and raised
– overlapping rings
– hair loss
 Ringworm can look different depending on which part of the body is affected.
Doctors call ringworm different names depending on where it appears on the body.
 Body  The term “ringworm” is most commonly used to refer to tinea corporis, or ringworm of the
body. This form often appears as patches with the characteristic round ring shape on your torso or limbs.

68
PAGE NO
 TA SLIDES BOOKSERIES

 Scalp  Ringworm of the scalp, or tinea capitis, often starts as isolated scaling in the scalp that
develops into itchy, scaly bald patches. It‟s most common among children. Hair around the affected area
may break or fall off, and bald patches may develop.
 Beard  Ringworm of the beard, also called tinea barbae, affects your cheeks, chin, and upper neck
and can cause bald patches. This may look like acne, folliculitis, or another skin condition. Some people
experience fatigue or swollen lymph nodes.
 Hands  Ringworm of the hand, or tinea manuum, is
usually caused by touching another affected area, such as your
groin or foot. Infection of the hand may look like very dry skin
with deep cracks on the palm.
 If the infection spreads, you may see ring-shaped patches on
the back ofyour hand.
 Groin  Jock itch, known as tinea cruris, refers to ringworm infection of the
skin around the groin, inner thighs, and buttocks. It‟s most common in men and
adolescent boys.
 This usually starts as an itchy red, brown, or gray rash where your leg and body
meet. The itching may intensify after exercise and may not improve after using
anti-itch cream.
 Feet  Athlete‟s foot, or tinea pedis, is the common name for ringworm
infection of the foot. It‟s frequently seen in people who walk barefoot in public
places where the infection can spread, such as locker rooms, showers, and swimming
pools.
 This starts as dry scaly skin between your toes that may spread to your sole and
heel. Symptoms may include:
– an itching, stinging, or burning sensation – blistering ]
– peeling – a foul odor
 Nails  Onychomycosis, also called tinea unguium, is a ringworm
infection of the nails. It affects toenails more than fingernails, as footwear often
provides a moist, warm environment that fungi prefer.
 Affected nails may become thicker or discolored. They may even begin to
crack or lift away from your nail bed.
Causes of ringworm
 About 40 different species of fungus can cause ringworm. They are typically of the Trichophyton,
Microsporum, and Epidermophyton types.
 These fungi can live on your skin and other surfaces, particularly damp areas. They may also live for an
extended period of time as spores in soil.
The fungi can spread to humans in four ways:
 Human to human. You can get the infection if you come in contact with a person who has ringworm
or if you share personal items, such as combs or towels. The infection is commonly spread among children
and by sharing items harboring the fungus.
 Animal to human. You can get ringworm after touching an affected animal or even items the animal
has come in contact with. Cats and dogs are common sources, but other animals, such as farm animals, can
spread the fungi as well.
 Object to human. You may get the infection if you come in contact with an object or surface that has it,
such as a telephone or the floor of a public shower. These fungi thrive in damp environments.
 Soil to human. Humans and animals can get ringworm after direct contact with soil that is carrying the
fungi.
Ringworm risk factors
 Anyone can get ringworm, but you may be more at risk if you:
– live in a warm, humid environment or climate – participate in contact sports, like wrestling or football
– use public showers or locker rooms – come in close contact with animals
– wear tight shoes or clothes that chafe your skin – have diabetes
– have obesity or are overweight – sweat excessively
– have a weakened immune system
Getting a ringworm diagnosis
69
PAGE NO
 TA SLIDES BOOKSERIES

 Your doctor will diagnose ringworm by examining your skin and possibly using a black light to view
the affected area. Depending on the type of fungus, it may sometimes fluoresce (glow) under black light.
 Your doctor may confirm a diagnosis of ringworm by requesting certain
tests: – If you‟re getting either a skin biopsy or fungal culture, your doctor will take a sample of your skin or
discharge from a blister and send it to a lab to test for the presence of fungus.
– If you‟re getting a KOH exam, your doctor will scrape off a small area of affected skin onto a slide and
place drops of a liquid called potassium hydroxide (KOH) on it. The KOH breaks apart typical skin cells,
making the fungal elements easier to see under a microscope.

Ringworm treatment
 Your doctor may recommend both medications and lifestyle adjustments to treat ringworm.
 Medications
 Your doctor may prescribe various medications depending on the severity of your ringworm infection.
 Jock itch, athlete‟s foot, and ringworm of the body can all typically be treated with topical medications,
such as antifungal:
– creams – ointments – gels – sprays
 Some severe cases may be treated with oral medications.
 Ringworm of the scalp or nails may require prescription-strength oral medications, such as griseofulvin
(Gris-PEG) or terbinafine.
 Your doctor may recommend over-the-counter (OTC) medications and antifungal skin creams as well.
These products may contain clotrimazole, miconazole, terbinafine, or other related ingredients.
Lifestyle adjustments
 In addition to prescription and OTC medication, your doctor may recommend that you care for the
infection at home by:
 Washing bedding and clothing daily to help disinfect your surroundings
 Drying your skin thoroughly after bathing  Wearing loose clothing in affected areas
 Treating all affected areas, as not treating tinea pedis can lead to the recurrence of tinea cruris

NOTE: IF YOU FOUND ANY MISTAKE CORRECT IT AND REPORT AT sta1372002@gmail.com

70
PAGE NO
 TA SLIDES BOOK SERIES
Unite 07 Microbiology in Everyday Life
What is Airborne Microorganism
Definition of Aero-microbiology:
‘Study of living microbes suspended in air’
 Number of microorganism in atmosphere but air is not
a natural environment for microorganism because it does
not contain enough moisture and nutrient.
 Airborne microbes are biological airborne
contaminants (also known as bioaerosois) like bacteria,
virus or fungi as well as airborne toxins passed from one
victim to the next through the air, without physical contact.
 Bacteria are emitted from the human respiratory
airways through talking,coughing,or sneezing. They also
get into the air with skin scales.
Water-borne Infections
 Infections in which the enteric microorganism enter the water source through faecal contamination and
transmission occur by the ingestion of contaminated water.
 Route of transmission and infection depend on:
 Amount of faecal contamination in water
 Concentration of pathogens in the faecal contamination
 Individual health status
 Control : Improvement of microbiological water quality(water treatment or Source protection).
 Example : Typhoid and cholera
How Do Infectious Microorganism Contaminate Drinking Water
 Many infectious microorganism found in the environment ,bacteria (such as shigella,E-coli,vibrio,and
Salmonella),Viruses (such as Norwalk virus and rotaviruses),and protozoans(such as
Entamoeba,Giardia)may be found in water.
 Infectious microorganism may be present in human or animal waste. Wells and other drinking water
sources can be contaminated by storm water run off from roadways,farms,and livestock operation,
discharge from sewage treatment plants, or septic system discharges.
Drink properly Treated water
 Do not drink untreated water from lakes,rivers,springs,ponds, Streams, or shallow wells.
 Do not drink tap water or use ice while travelling to a high –risk destination unless the water sources
has been properly treated.
 Drink only filtered/boiled water.
 Store water in a clean container.
 Water jars/containers should be washed daily.
Common Airborne, Waterborne, and Foodborne Diseases
Airborne: Water-Borne Food Borne
Covid-19 Polio Botulism
Common cold Malaria E-Coli infection
Influenza Cholera Salmonellosis
Chickenpox Dengue Shigellosis
Mumps Scabies Hepatitis A
Measles Typhoid listeriosis
TB /pertussis Hepatitis/Diarrhea Norwalk/Rota virus
How Microorganism Affects
 The primary harmful effects of microbes upon our existence and civilization is that they are an
important cause of disease in animal and crop plants, and they are also agents of spoilage and decomposition
of our food ,textile and dwellings.
 Microbes cause infectious diseases in human such as flu and measles. There is also strong evidence that
microbes may contribute to many non infectious chronic diseases such as some form of cancer and Coronary
heart diseases.
 Different disease are caused by different microorganism.

71
PAGE NO
 TA SLIDES BOOK SERIES
Disease-Causing Microbes in Animals
 Several microorganism not only cause diseases in humans and plants,
but also in other animals. For example ,anthrax is a dangerous human and
cattle disease caused by bacterium. Foot and mouth disease of cattle is
caused by virus.
 Robert Koch (1876) discovered the bacterium (bacillus anthracis)
which cause anthrax disease.
Disease Causing Microbes in Plants
 Several microbes cause disease in plants
like,wheat,rice,potato,sugarcane,orange,apple,and others. This
diseases reduce the yield of crops. They can be controlled by the
use of certain chemicals which kill the microbes.
Harmful Microbes
 Microbes are harmful in many ways. Some of the microbes
causing diseases in human beings,plant,and animals. Such
disease-causing microorganism are called pathogen.
 Pathogen enter our body through the air we breath, the water
we drink or the food we eat. They can also get transmitted by
direct contact with an infected person or carried through an animals. Microbial disease that can spread from
an infected person to a healthy person through air ,water, physical contact are called communicable diseases.
 Example of such diseases include cholera, common cold,chiken pox, and TB.
 When a person suffering from common cold sneezes, fine droplets of moisture, carrying thousands of
viruses are spread in the air. The virus may enter the body of healthy person while breathing.
 There are some insects and animals which act as carrier of disease-causing microbes.
 Housefly is one such carrier.
 The fly sit on the garbage and animals excreta. Pathogen stick to their bodies. When these flies sit on
uncovered food they may transfer the pathogen.
 Whoever eats the contaminated food is likely to get sick.
 Example of carrier are: the female anopheles mosquito which carries the parasite of malaria and female
aedes mosquito acts as carrier of dengue virus.
Some Common Plant Diseases Caused by Microbes
Plant disease Microbe Mode of transmission
Citrus canker Bacteria (xanthomonas sp.) Air
Rust of wheat fungi (Puccinia triticina) Air, seeds
Yellow vein mosaic of bhindi Virus (begomovirus) Insect
(okra)
Human Diseases Causative microbes Mode of transmission
TB Bacteria Air
chicken pox Virus Air/Contact
Measles Virus Air/Contact
Polio Virus Air/Water
Cholera Bacteria Water/Food
Typhoid Bacteria Water
Hepatitis A Virus Water
Malaria Protozoa Mosquito
Preventive Measures:
 Keep the patient in complete isolation.
 Keep the personal belonging of the patient away from those of others. Vaccinations to be given at
suitable age.
 Maintain personal hygiene and good sanitary habits. Consume properly cooked food and boiled
drinking water. Vaccination
 Drink boiled drinking water. Vaccination
 Use mosquito net and repellents. Spray insecticides and control breeding of mosquito by not allowing
water to collect in the surrounding.
 Health education programs across the country.  Careful disposal of all waste products.
72
PAGE NO
 TA SLIDES BOOK SERIES
Food Spoilage
 Food spoilage is the state of food where it is no longer safe to consume.
 As soon as food is harvested, slaughtered or manufactured into a product it starts to
change. This is caused by two main processes:
 Autolysis – self destruction, caused by enzymes present in the food;
 microbial spoilage – caused by the growth of bacteria, yeasts and moulds.
Enzymes & Chemical Reactions
 Enzymes help facilitate chemical reactions that cause foods to spoil.
 As Chemicals in food are altered, the smell, appearance, texture and
flavor can change.
 ex. Oxidation of apples and potatoes when sliced; In apples, the specific
enzyme that causes the brownish color to appear is called polyphenol oxidase
(PPO).
Food Spoilage
 Any sensory change (visual,olfactory,or flavor) which the consumers considers to be unacceptable.
 The main single cause of food spoilage is invasion by microorganism such as moulds,yeast and bacteria.
 The presence of highly dangerous toxins and bacterial spores is often not detected until after an
outbreak of food poisoning,labortory examination uncovers the infecting agent.
How Do Microorganisms Spoil food
 Microbial spoilage is caused by microorganism like fungi,(moulds,yeast) and bacteria.
 They spoil food by growing in it and producing substances that change the
color,texture,and odor of the food.
 Eventually the food will be unfit for human consumption.
SPOILAGE !!!! Remember:
When in doubt, throw it out.
Factors that Promote Enzymes and Microbial Activity
Micro-organisms and enzymes need certain conditions to survive and reproduce.
These include:
• Temperature. • Oxygen. • moisture.
• pH level.
Food Poisoning
 Food Poisoning is an illness caused by eating contaminated food.
 The food is contaminated by bacteria, such E-coli, salmonella, or virus, such
as norovirus.
 Microbes that grow in our food sometime produce toxic substances. These make the food poisonous
causing serious illness and even death.so it is very important that we preserve food to prevent it from being
spoilt.
Staphylococcal Food Poisoning
 Staphylococcus aureus-gram positive microorganism.
 Human harbor these organism ,present in boils, skin lesions.
 Food poisoning is caused when staph is transferred from person to food (could come from an open
boil,lesions,sneeze,etc.)
 Exotoxins are heat stable protein known as enterotoxins (affecting the intestine).
 Inhibit water absorption from the intestine and induce diarrhea.
 Demage the intestinal lining-colitis.  Nausea,vomitting abdominal cramps ,diarrhea.
 Onset-1-6 hours after eating the contaminated food.
Clostridium Botulinum& Botulism
 C.Botulinum gram positive, endospore forming organism, anaerobic.
 Present in soil, ponds and lakes.  Infrequent, but fatal.
 Caused by exotoxins produced by C.botulinum, or neurotoxin and affects the CNS.
Clostridium Perfringes
 Anaerobic spore forming microorganism.
 Present in soil and intestine of humans and animals hence also in sewage.
 Typical illness- Diarrhea, abdominal cramps, nausea, vomiting.
 Incubation period is usually 6-24 hours.
73
PAGE NO
 TA SLIDES BOOK SERIES
 Prevention-refrigerate food and cook adequately.
Foodborne Diseases
 Usually called ‘food poisoning, caused by eating the food contaminated with toxin.
 Two main types:
 Toxin is produced in the food (exotoxin) caused the diseases called food poisoning.
 Food, being a mechanical carrier, carriers pathogens and cause disease in ingestion.
Foodborne Pathogens
 Gram negative bacteria:
 Salmonella  Shigella  Vibrio Cholera
 Amoeba:
 Giardia lamblia, Entamoeba histolytica, cryptosporidium.
 Fungi:
 Aspergillus-aflatoxins, Claviceps-halluciogens.
 Algae:
 Certain marine algae produce neurotoxins contaminate shell fish.
Food Borne Infection
 Diseases caused by microorganism through food.
 Food borne infections are caused by the entrance of pathogenic, microorganism contaminating food into
the body, and the reaction of the body tissue to their presence.
 These can either be fungi,bacteria,viral,parasite,or protozoa.
 Microorganism can cause food spoilage, diseases.
Bacterial foodborne infection:Include,salmonellosis,typhoid fever, cholera, E-coli
infection,shigellosis,yersiniosis, campylobacteriosis.
Mycotic foodborne infection: Include candida spp,sporothrix spp,wangiella spp,etc.
Viral foodborne infection: Include hepatitis A&E, Norwalk, and poliomyelitis virus.
Parasite foodborne infection: Include anisakiasis, trichinosis, taeniasis, hydatidosis.
Protozoal foodborne infection: Include,crptosporidiosis,taxoplasmosis,sarcocystosis,and cyclosporiasis.
Different Between Food Infection and Food Poisoning
Infections Intoxications
Bacterial /viral/parasite/protozoa Toxins(natural/ preformed bacterial/ chemical
Invade and /or multiply in lining of Intestine No invasion or multiplication
Incubation period-hours to days Incubation period minutes to hours
S/S- Diarrhea, nausea, vomitting Abdominal S/S- vomitting, nausea, diarrhea, weakness,
cramps, fever sensory/motor/dysfunction
Communicable spread from Person to person Not communicable
Cause infection Toxin is the direct cause cause of disease
Transmission of Foodborne Diseases
 Four main ways , 4Fs
 Food  Feces  Finger  Flies
 Food is the immediate source of the pathogen /toxin.
 May be acquired by close contact with an infected person or a carrier
 Involves the fecal –oral route.
Prevention of Food and water Borne illness
 Wash and/or peel all raw vegetable and fruits before eating.
 Drink and eat only pasteurized dairy products (milk,cheese,yogurt,and ice cream) and juices.
 Wash hands,kichen work surface and utensils after contact with raw meat or poultry.
 Wash hand before handling food and between handling different food items.
 Prevent contact of cooked foods with raw foods (i.e. raw meat and poultry)
 Clean and sanitize all utensils, equipment and surface (cutting boards,
work counters,etc.) before and after each use.
 Be sure to use hot water and detergent to clean ,then rinse with hot
water .Sanitize food contact surfaces with a sanitizing solution.
 Avoid preparing food for other while you have symptoms and for 48 hours
after you recovers.

74
PAGE NO
 TA SLIDES BOOK SERIES

Food Preservation
 Food preservation is the process of treating and handling food in such a way as to stop or greatly slow
down spoilage to prevent foodborne illness and extend its shelf-life while keeping as much of its nutritional
quality.
 We add salt or acid preservative to pickles to prevent the attack of
microbes.
 Sodium benzoate and sodium metabisulphite are common preservatives.
These are also used in jams and squashes to check their spoilage.
 Common salt has been used to preserve meat and fish for ages. Meat
and fish are covered with dry salt to check the growth of bacteria. Salting is
also used to preserve amla,raw mangoes etc.
 Jams ,jellies, and squashes are preserved by sugar. Sugar reduces the moisture content which inhibits
the growth of bacteria which spoil food.
 Use of oil and vinegar prevent spoilage of pickles because bacteria can not live in such an
envirnment.vegetable ,fruits,fish,and meat are often preserved by this method.
Freezing
 Reducing the temperature of the food to below – 18ºC reduces the activity
of the micro-organisms and enzymes.
 Freezing also reduces the availability of water because ice crystals are
formed.
 In China, freezing has been a method of preservation for hundreds of
years, dating back to 1800 BC.
Dehydration
 The term ‘dehydration’ means the progressive reduction of water
content inside foods with the aim of inhibiting microbial spoilage;
essentially, this process has micro biostatic effects.
 Dehydration/drying stops/slows : respiration, enzyme activity and
microbial activity
 Sun drying, heat drying, freeze drying.
Pasteurization
 Boiling kills many microbes in milk. Low temperature inhibits the growth of microbes.
 Pasteurized milk can be consumed without boiling as it is free from harmful microbes.
The milk is heated to about 70 degree Celsius for 15 to 30 seconds and then suddenly chilled
and stored.
 By doing so, it prevent the growth of microbes.it is called pasteurization.
Methods of food preservation - Pickling
 The initial boiling of the ingredients will destroy enzymes and micro organisms (but not spores),
preventing spoilage later on.
 Vegetables and fruits are covered in vinegar and other ingredients, often including spices. The high
concentration of acid inhibits bacterial growth and multiplication. The acidic nature of the solution prevents
growth of micro-organisms.
 Pickle/or chutney jars are normally heated before the product is added to destroy micro-organisms
found in the jars.

75
PAGE NO
 TA SLIDES BOOK SERIES
USE OFMICROSCOPE
What is a Microscope?
 MICROSCOPE (Greek: mikron = small and scopeos = to look): Is an instrument for
viewing objects that are too small to be seen by the naked or unaided eye.
 Microscope is an optical instrument that uses a lens or combination of lenses to produce
magnified images of small objects, especially objects too small to be seen by the unaided
eye.
 MICROSCOPY: The science of investigating small objects using such an instrument is
called microscopy
History of Microscope
 Light microscope is one of the most basic and essential pieces of equipment used in any
laboratory.
 It is used for visualizing very small objects like cells, bacteria, parasite, Ova/cyst etc.
 1590 - Hans Janssen and his son Zacharias Janssen, developed first microscope.
 1667: Robert Hooke's famous "Micrographia" is published, which outlines Hooke's
various studies using the microscope
Basic Terminology
 MAGNIFICATION-Degree of enlargement: No
of times the length, breadth or diameter, of an object
is multiplied.
 TOTAL MAGNIFICATION: magnification of
the eyepiece x magnification of the objective.
 RESOLUTION: Ability to reveal closely
adjacent structural details as separate and distinct.
Microscope

How to Operate
• 1.The microscope should be placed on a level bench, which should be free of vibration.
• 2.The power socket to which the microscope is plugged should not be loose or sparking.

PAGE NO
76
 TA SLIDES BOOK SERIES
• 3.The height of the microscope or chair should be adjusted in such a way that the users eyes
are directly on the eyepieces.
• 4.Place the specimen on the stage ,switch to the x10 objective and focus.
• 5.Turn on the power at the wall and the power switch on the microscope.
• 6.Ensure the light beam diaphragm is opened.
How to Operate
• 7.Start with the light setting at low to medium.
• 8.Depending on the sample being examined, the condenser may need to be adjusted.
• 9.Secure the slide using the arms on the microscope stages.
• 10.When starting to examine a slide, select the low powered(X4) object lens first.
• 11.Your eye level should be just above the eye pieces then look down the eye pieces and
gently slide them together until you see a single object.
• 12.Locate an area of the slide that includes part of the sample .
• 13.To bring the slide into focus rotate the coarse focus slowly until the sample can be
visualized clearly.
• 14.Once an area of interest has been identified, rotate through the objective lenses to a
higher power.
• 15.The oil immersion lens(and oil) is required to look at cell detail or bacteria.
• 16.Rotate the objective lenses slightly leaving a space to add a drop of immersion oil to the
slide, directly over the spot of light.
• 17.Look down the eyepieces and use the fine focus to bring the image into focus.
• 18.Once the examination is completed, remove the slide and clean the oil from the
immersion lens. Only use lens tissue on microscope objectives.
Application of Light Microscopy
 Light microscopes play a large part in today’s biology.
 Microscopes are used in biology to study diseases like cancer and AIDS to help
diagnose the disease in patients and to help find a cure for them.
 This device is used for observing microorganisms and their features. In this field,
microscopes are used to study bacteria, cells and many more. This device helps biologists in
their study of living organisms and their cell structures.
 The main application of microscopes is scientific research. Mineralogists also use
light microscopy, typically with a special preparation of a sample called thin section.
 As the name implies, thin section are very thin slices of a rock. The sample needs to be
thin enough for light to travel through from the light source to the user’s eye.
 The thin section will allow the shape of different crystal grains to be seen

PAGE NO
77
 TA SLIDES BOOK SERIES
Gram Staining procedure
Introduction
Stained preparations are needed to examine micro-organisms microscopically in order to study their
morphology and
observe their cellular constituents.
Smears (or tissue sections) are made and stained by any one of the required staining methods.
Smears can be made from liquid or solid cultures or from the clinical specimen.their
morphology and observe their cellular constituents.
Staining and Stain
Staining: procedure that applies colored chemicals called dyes to the specimen in order to facilitate
identification.
Stains: salt composed of a positive and negative ions, one of which is colored (chromophore color being
ions), which impart a color to cell or cell parts and fixed to them through a chemical reaction.
GRAM POSITIVE VS GRAM NEGATIVE BACTERIA
Most bacteria can be broadly classified as Gram +ve or Gram -ve.
Gram +ve bacteria have cell walls composed of thick layers of peptidoglycan.
Gram +ve cells stain purple when subjected to a Gram stain procedure.
Gram -ve bacteria have cell walls with a thin layer of peptidoglycan. The cell wall also includes an outer
membrane with lipopolysaccharide (LPS) molecules attached.
Gram -ve bacteria stain pink when subjected to a Gram stain procedure.
GRAM POSITIVE VS GRAM NEGATIVE CELL WALL

Types Of Staining

Gram Staining
The name come from the Danish bacteriologist Hans Christian gram, who developed this technique.
Gram staining is used to distinguish gram positive and gram negative bacteria.

78
 TA SLIDES BOOK SERIES
Smear Preparation
To stain material from a culture growing on solid media, place a loop full of tap water on a slide, using a
sterile cool loop transfer a small sample of the colony to the drop, and emulsify.
Allow the film to air dry.
(morphology and
observe their cellular
constituents.
Smears (or tissue
sections) are made
and stained by any
one of the required
staining methods.
Smears can be made from liquid or solid cultures or from
the clinical specimen Stained preparations are
needed to examine micro-organisms microscopically in
order to study their morphology and observe
their cellular constituents.
Smears (or tissue sections) are made and stained by any
one of the required staining methods.
Smears can be made from liquid or solid cultures or from
the clinical specimen)
NOTE ABOVE TOPIC HIDE BEHIND PICTURE IN SLIDES

POINT TO BE NOTED :

Staining Steps

79
 TA SLIDES BOOK SERIES
Flood the slide with crystal violet solution for up to one minute. Wash off briefly with tap water. (Not
over 5 second).
Flood slide with gram iodine solution, and allow to act for one minute. Wash off with tap water.
Flood slide with 95% alcohol for 10 second and wash off with tap water.
Drain the slide.
Flood slide with Safranine(counterstain) solution and allow for 30second.wash up with tap water.
All slides of bacteria must be examined under the oil immersion lens.
G+ G-

80
 TA SLIDES BOOK SERIES
SAFE USE OF LAB
Why is Lab Safety Important?
• Lab safety rules and symbols are needed so that students do not injure themselves or their classmates.
Main Laboratory Hazards and Accidents
• Infections • Burns • Harmful effects of toxic chemicals
• Injury from explosions • Electric shock

INFECTIONS
• Laboratory-acquired infections (LAIs) are defined as all infections acquired through laboratory or
laboratory-related activities regardless of whether they are symptomatic or asymptomatic in nature.
INHALATION
 Aerosol
• Centrifugation in open buckets
• Breakage in centrifuge
INGESTION
• Contaminated fingers
• Food stored in Lab. Refrigerator
• Mouth pipetting
ENTRY INTO THE BLOOD
 Pathogens entering the skin through:
• Needle punctures (Needle Stick Injury) • Cuts
• Scratches • Insect Bites
• Sores or other open skin lesions
BURNS
 Burn Injuries can occur through:
• Flammable chemicals and stains
• Fire from spirit lamp, (spirit burner should not be used in direct sunlight because in direct light flame can
be difficult to see).
• Corrosive chemicals being spilt on the skin or ingested when mouth pipetting.
CUTS
• Intact skin is the first line of defense.
• Skin breakage can occur while using a sharp blade.
• Using glassware that is cracked or has damaged Edges
.HARMFUL EFFECTS OF TOXIC CHEMICALS
• Inhaling fumes from toxic chemicals. • Ingesting toxic chemicals by mouth pipetting.
• Skin contact with toxic chemicals.
INJURY FROM EXPLOSIONS
• Incompatible chemical exploding. • Leak gas exploding.
ELECTRIC SHOCKS
• Faulty electric circuits. • Incorrect installation of equipment.
• Touching exposed live wires.
FACTORS CONTRIBUTING TO LAB. ACCIDENTS
• Poor training • Lack of concentration • Noisy environment
• Untidy working and not using racks to hold containers • Allowing the working bench to become cluttered
• Carelessness and negligence • Hurry to finish workLab Safety Symbols
81
 TA SLIDES BOOK SERIES
• They alert about the possible dangers in the lab.

.Lab Safety Rules

1. Wear protective clothing
Protective clothes
o Gloves are essential.
o Lab coats are required.
o Safety glasses ( goggles) may be required to avoid splashes
2. Laboratory personnel should not wear sandals
• Do NOT Wear:

• Jewelry
• • Loose or Baggy clothing
3. Avoid touching objects (e.g., pencils, cell phones, door handles) while wearing gloves.
4. Pencils, labels, or any other materials should never be placed in your mouth.
5. Caution must be taken when using gas burners. Be sure gas burners are turned off when finished.
6. Long hair must be tied back or covered to minimize fire hazard or contamination of experiments.
7. Do not eat food or drink water in the lab. do not use lab glassware as food or water containers.
8. Protect your hands:
 Wash hands after every lab.  Handle glassware, sharp tools and heated containers carefully.
9. Electrical safety:
 Unplug electrical equipment after use.  keep all electrical cords and wires away from water.
10. Chemical safety:
 Never touch, taste or smell a chemical unless instructed to do so.
 Never mix chemicals unless instructed to do so.
 Keep lids on chemical containers when not in use.
11. Do not take any cultures out of the lab for any reason.
 All cultures should be handled as potentially pathogenic.
 Liquid cultures must always be kept in a test tube rack.
12. Do not engage in practical jokes or horseplay in the lab.
13. Keep nonessential books and clothing far away from your work area.
14. Wipe the bench tops down with disinfectant both before you begin your work and after you have completed
your work.
15. Dispose of waste products according to instructions.
82
 TA SLIDES BOOK SERIES
16. Report all accidents, no matter how
minor, to your supervisor!!
Lab Safety Equipment
• Safety Shower
• Shower Eye washer
• Fire Blanket • Fire Extinguisher

Biological Safety Levels

• 1. BSL1 • 2. BSL2 • 3. BSL3 • 4. BSL4

BSL-1
• Not known to cause disease in healthy adult humans
 Safety equipment  Minimal requirements  Facilities  Open bench top
83
 TA SLIDES BOOK SERIES
BSL-2
• Associated with mild to moderate disease in humans
o Safety equipment
 Biological Safety Cabinet and personal protective equipment as needed.
o Facilities
 BSL-1 plus the availability of a mechanism for decontamination.
BSL-3
• Associated with serious or potentially lethal disease in humans.
 Safety equipment  Biological Safety Cabinet and personal protective equipment required.
 Facilities  BSL-2 with self-closing double door access and single-pass negative directional airflow.
BSL-4
• Associated with high risk of life-threatening disease in humans and/or animals.
 Safety equipment
 Biological Safety Cabinet Full-body air-supplied, positive pressure personnel suit
 Facilities
 BSL-3 plus dedicated air and exhaust, decontamination procedures for exit, separate building, etc.

84
 TA SLIDES BOOK SERIES
CULTUREMEDIA
Culture/Medium Defined
• CULTURE : Is the term given to microorganisms that are cultivated in the lab for the purpose of
identifying and studying them.
• MEDIUM: Is the term given to the combination of ingredients that will support the growth and
cultivation of microorganisms by providing all the essential nutrients required for the growth
(i.e.multiplication) in order to cultivate these microorganisms in large number to study them.
• Inoculation: Introduction of microbes into the medium.
Reasons for bacterial cultural
• Subsequent clinical diagnosis.
• Studying its morphology and its identification.
• Bacteria have to be cultured in order to obtain antigens from developing serological assay for vaccines.
• Certain genetic studies and manipulations of the cells also need that bacteria be cultured in vitro.
The Requirements for Bacterial Growth
 Physical Requirements:
• Temperature • pH • Osmotic Pressure
 Chemical Requirements:
• Carbon • Nitrogen • Phosphorous
• Oxygen • Hydrogen • Trace Elements
Types of Media
 Chemically Defined Media: Exact chemical composition is known.
 Complex Media: Extracts and digests of yeasts, meat, or plants.
Types of Media
A Chemically Defined Medium for Growing a Typical Chemoheterotropc, such as E.coli
Constituent Amount
Glucose 5.0g
Ammonium phosphate, (NH4H2PO4) 1.0g
Sodium Chloride (NaCl) 5.0g
Magnesium Sulfate (MgSO4 - 7H20) 0.2g
Potassium Phosphate, (K2HPO4) 1.0g
Water 1 liter
Composition of Nutrients, a Complex Medium for the Growth of Heterotrophic Bacteria
Constituent Amount
Peptone (Partially digested protein) 5.0g
Beef extract 3.0g
Sodium Chloride 8.0g
Agar 15.0g
Water 1 Liter
Types of Culture Media
 Selective Media:
• Suppress unwanted microbes and encourage desired microbes.
 Differential Media:
• Make it easy to distinguish colonies of different microbes.
 Enrichment Media:
• Encourages growth of desired microbes.
Examples of Selective Media
• Blood agar • MacConkey agar
• Hektoen enteric agar (HE) • Mannitol salt agar (MSA)
Examples of Differential Media
• Eosin methylene blue (EMB) • MacConkey agar
• Mannitol salt agar (MSA)
1
85
 TA SLIDES BOOK SERIES
Examples of Enriched Media
• Chocolate agar • Blood agar
Blood Agar
• Contains 5-10% mammalian blood (usually sheep or
horse)
• Contains meat extract, NaCl, and agar
• Used to detect hemolytic activity
• Alpha-hemolysis- partially lysis of RBC
• B-hemolysis- Complete lysis of RBC
• Y-hemolysis- no lysis
MacConkey Agar
• It contains bile salts (to inhibit Gram-positive bacteria)
• Crystal violet dye
• Neutral red dye (which stains microbes fermenting lactose)
• Lactose
• Peptones
Mannitol Salt Agar (MSA)
• It contains a high concentration (7.5- 10%) of salt (NaCl), making it selective for
Staphylococci and Micrococcaceae
Eosin Methylene Blue
• Is a selective stain for Gram-negative bacteria
• Inhibits the growth of Gram-positive bacteria
and provides a color indicator distinguishing
between organisms that ferment lactose (e.g.,
E.coli) and those that do not (e.g., Salmonella,
Shigella).
Chocolate Agar
• Is a non-selective, enriched growth medium
• Contains red blood cells, which have been lysed by
heating
Lowenstein-Jensen medium
• For the cultivation and differentiation of
Mycobacterium species
Mueller-Hinton Agar (MHA)
• For antimicrobial susceptibility testing

2
86