Assignment –Micribiology & Medical care

Q 1:- Malaria parasites can be identified by examining under the microscope a drop of the
patient's blood, spread out as a “blood smear” on a microscope slide. Prior to examination,
the specimen is stained (most often with the Giemsa stain) to give the parasites a distinctive
appearance.

Microscopic examination remains the “gold standard” for laboratory confirmation of malaria.
These tests should be performed immediately when ordered by a health-care provider. They
should not be saved for the most qualified staff to perform or batched for convenience. In
addition, these tests should not be sent out to reference laboratories with results available
only days to weeks later. It is vital that health-care providers receive results from these tests
within hours in order to appropriately treat their patients infected with malaria.

Technique
(See DPDx specimen preparation) A blood specimen collected from the patient is spread as a
thick or thin blood smear, stained with a Romanovsky stain (most often Giemsa), and
examined with a 100X oil immersion objective. Visual criteria are used to detect malaria
parasites and to differentiate (when possible) the various species.
(See DPDx Plasmodium species comparison chart) Wright’s stain, which is commonly used in
hospital laboratories for examining blood (called a CBC with manual differential), can be used
if Giemsa stain is not available. However, species determination might be more difficult.

Blood smear from a patient with malaria; microscopic examination shows Plasmodium
falciparum parasites (arrows) infecting some of the patient’s red blood cells. (CDC photo)

Advantages
Microscopy is an established, relatively simple technique that is familiar to most
laboratorians. Any laboratory that can perform routine hematology tests is equipped to
perform a thin and thick malaria smear. Within a few hours of collecting the blood, the
microscopy test can provide valuable information. First and foremost it can determine that
malaria parasites are present in the patient’s blood. Once the diagnosis is established –
usually by detecting parasites in the thick smear – the laboratorian can examine the thin
smear to determine the malaria species and the parasitemia, or the percentage of the
patient’s red blood cells that are infected with malaria parasites. The thin and thick smears
are able to provide all 3 of these vital pieces of information to the doctor to guide the initial
treatment decisions that need to be made acutely.

Disadvantages
Microscopy results are only as reliable as the laboratories performing the tests. In the United
States, there are, on average, 2000 cases of malaria diagnosed and reported each year. Thus,
the average laboratorian does not perform this test regularly, and may not be maintaining
optimal proficiency.

Q 2:- Basal media also called general-purpose media are basically simple media that
support the growth of most non-fastidious bacteria. Peptone water, nutrient broth, and
nutrient agar (NA) are considered basal media. These media are generally used for the
primary isolation of microorganisms. BASAL MEDIA. Basal media are those that may be
used for growth (culture) of bacteria that do not need enrichment of the media.
Examples: Nutrient broth, nutrient agar and peptone water. Staphylococcus and
Enterobacteriaceae grow in these media.
Simple Media is a multi-disciplined creative agency with a focus on website development,
social media strategy and management. The basal medium is prepared by anaerobic
incubation of all ingredients in RGCSA medium except the carbohydrates, Na2CO3, and
cysteine for 7 days at 38 degrees C. After incubation, substrate(s), Na2CO3 and cysteine are
added and the medium is tubed and sterilized as in normal medium preparation. Fetal bovine
serum (FBS) is a byproduct of harvesting cattle for the meatpacking industry—it's used
extensively by both academic and industrial researchers as a supplement to basal growth
medium in cell culture applications.

Q3:- An elevated titer of antibody (positive ASO) or an ASO titer that is rising means that it
is likely that the person tested has had a recent strep infection. ASO titers that are initially
high and then decline suggest that an infection has occurred and may be resolving.
You'll need to give a blood sample for an ASO titer test. A nurse or lab technician will take a
blood sample from a vein in your inner arm or hand. They'll use a needle to enter your vein
and draw your blood into a tube. Then they'll send this tube to a lab for analysis. An
antistreptolysin O titer (ASO) is a blood test used to determine if you've had a recent
infection caused by group A streptococcus bacteria. Principle. The detection of anti-
streptolysin O antibodies is based upon the neutralization of the streptolysin O hemolytic
activity by antibodies present in the test serum. The antigen-antibody immunological
complexes that are formed are revealed by the addition of a suspension of sheep
erythrocytes.
Generally, an ASO test value below 200 is considered normal. In children under the age of
5, the test value should be less than 100. Results will vary by laboratory. If your results show
that you have an elevated ASO value, you may have a post-streptococcal complication.

Q4:- The basic principle of gram staining involves the ability of the bacterial cell wall to
retain the crystal violet dye during solvent treatment. Gram-positive microorganisms have
higher peptidoglycan content, whereas gram-negative organisms have higher lipid content.
Simple staining involves directly staining the bacterial cell with a positively charged dye in
order to see bacterial detail, in contrast to negative staining where the bacteria remain
unstained against a dark background.
Principle of Negative Staining

Since the surface of most bacterial cells is negatively charged, the cell surface repels the
stain. The glass of the slide will stain, but the bacterial cells will not. The bacteria will show
up as clear spots against a dark background.
Procedure of Gram Staining
Flood the gram's iodine for 1 minute and wash with water. Then ,wash with 95% alcohol or
acetone for about 10-20 seconds and rinse with water. Add safranin for about 1 minute and
wash with water. Air dry, Blot dry and Observe under Microscope. Gram staining is a
common technique used to differentiate two large groups of bacteria based on their different
cell wall constituents. The Gram stain procedure distinguishes between Gram positive and
Gram negative groups by coloring these cells red or violet.
Differential staining techniques commonly used in clinical settings include Gram staining,
acid-fast staining, endospore staining, flagella staining, and capsule staining.
The performance of the Gram Stain on any sample requires 4 basic steps that include
applying a primary stain (crystal violet) to a heat-fixed smear, followed by the addition of a
mordant (Gram's Iodine), rapid decolorization with alcohol, acetone, or a mixture of alcohol
and acetone and lastly, counterstaining with

Q 5:- Principle of Moist Heat sterilization

Moist heat destroys microorganisms by the irreversible denaturation of enzymes and
structural proteins. The temperature at which denaturation occurs varies inversely with the
amount of water present.
The basic principle of steam sterilization, as accomplished in an autoclave, is to expose each
item to direct steam contact at the required temperature and pressure for the specified time.
Different methods are used to achieve sterilization. One of the most common is applying
moist heat which includes autoclaving (pressure cooking), boiling, and Tyndallisation. Dry
heat sterilization is accomplished by conduction and is used widely for instruments. Other
heat methods include flaming and incineration.
Moist heat sterilization is the sterilization technique using high-pressure steam. Based on the
temperature of sterilization ,moist heat sterilization is classified as : Temperature below 100
C Example: Pasteurisation ; Pasteurised milk. Moist heat sterilization is a procedure in
which heated, high-pressure steam is used to sterilize an object, killing any bacteria, viruses,
or spores that may have contaminated the object. ... Moist heat sterilization is performed in a
pressurized chamber called an autoclave.
The steam inside the autoclave is the agent by which the moist heat attacks the
microorganisms, thus causing them to coagulate and die.
 Q6:- Two types of immunity exist — active and passive: Active immunity occurs when our
own immune system is responsible for protecting us from a pathogen. Passive immunity
occurs when we are protected from a pathogen by immunity gained from someone else.
Innate immunity: Everyone is born with innate (or natural) immunity, a type of general
protection. ...
 Adaptive immunity: Adaptive (or active) immunity develops throughout our lives. ...
 Passive immunity: Passive immunity is "borrowed" from another source and it lasts for a
short time.
Explore the Different Types of Immunity
 Active Immunity. Active immunity is a type of immunity that is created by our own immune
system when we come in contact with a harmful pathogen. ...
 Passive Immunity. ...
 Innate Immunity. ...
 Adaptive Immunity.
There are two types of immunity: active and passive.
There are two types: passive and active. Passive immunity occurs when antibodies are
passed from one person to another, as through transfusion for example. The active immunity
involves two types of white blood cells - T-cells and B-cells.
Immunity refers to the body's ability to prevent the invasion of pathogens. Pathogens are
foreign disease-causing substances, such as bacteria and viruses, and people are exposed
to them every day. Antigens are attached to the surface of pathogens and stimulate an
immune response in the body.
Q7:- To diagnose ascariasis, your doctor will examine your stool for the tiny (microscopic)
eggs and larvae. But eggs won't appear in stool until at least 40 days after you're infected.
And if you're infected with only male worms, you won't have eggs. The standard method for
diagnosing ascariasis is by identifying Ascaris eggs in a stool sample using a microscope.
Because eggs may be difficult to find in light infections, a concentration procedure is
recommended. Ascariasis should be considered in the differential diagnosis when a patient
presents with pneumonia, peripheral eosinophilia, and/or intestinal obstruction in returned
traveler or resident from an endemic area. Ascariasis is an infection of the small intestine
caused by the parasitic roundworm Ascaris lumbricoides. Serologic test used for diagnosis
Ascaris infection.
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.
What causes an ascariasis infection? You can become infected with ascariasis
after accidentally ingesting the eggs of the A. lumbricoides roundworm. The eggs can be
found in soil contaminated by human feces or uncooked food contaminated by soil that
contains roundworm eggs.

Q8:- Bacteria are unicellular organisms belonging to the prokaryotic group where the
organisms lack a few organelles and a true nucleus”.

Bacteria Diagram
The bacteria diagram given below represents the structure of bacteria with its different parts.
The cell wall, plasmid, cytoplasm and flagella are clearly marked in the diagram.

Bacteria Diagram representing the Structure of Bacteria

Structure of Bacteria
The structure of bacteria is known for its simple body design. Bacteria are single-celled
microorganisms with the absence of the nucleus and other cell organelles; hence, they are
classified as prokaryotic organisms.
They are also very versatile organisms, surviving in extremely inhospitable conditions. Such
organisms are called extremophiles. Extremophiles are further categorized into various types
based on the types of environments they inhabit:

1. Thermophiles
2. Acidophiles
 3. Alkaliphiles
4. Osmophiles
5. Barophiles
6. Cryophiles

Another fascinating feature of bacteria is their protective cell wall, which is made up of a
special protein called peptidoglycan. This particular protein isn’t found anywhere else in
nature except in the cell walls of bacteria.
But few of them are devoid of this cell wall, and others have a third protection layer called
capsule. On the outer layer, one or more flagella or pili is attached, and it functions as a
locomotory organ. Pili can also help certain bacteria to attach themselves to the host’s cells.
They do not contain any cell organelle as in animal or plant cell except for ribosomes.
Ribosomes are the sites of protein synthesis. In addition to this DNA, they have an extra
circular DNA called plasmid. These plasmids make some strains of bacteria resistant to
antibiotics.
Also Read: Gram Positive Bacteria

Classification of Bacteria
Bacteria can be classified into various categories based on their features and characteristics.
The classification of bacteria is mainly based on the following:

 Shape
 Composition of the cell wall
 Mode of respiration
 Mode of nutrition

Q9:- A differential media contains specific ingredients or chemicals that allow the observer to
visually distinguish which species possess and which species lack a specific biochemical
process. Blood agar is a dExamples of differential media include: Blood agar (used in strep
tests), which contains bovine heart blood that becomes transparent in the presence of
hemolytic. Streptococcuseosin methylene blue (EMB), which is differential for lactose and
sucrose fermentation.ifferential medium that distinguishes bacterial species by their ability to
break down red blood cells. Differential media are used to differentiate closely related
organisms or groups of organisms. Owing to the presence of certain dyes or chemicals in the
media, the organisms will produce characteristic changes or growth patterns that are used
for identification or differentiation.
Blood agar is an enriched medium that provides an extra rich nutrient environment for
microbes. Therefore, BAP is not a selective growth medium, since it supports the growth of a
wide range of organisms.
Selective media are used to select for the growth of a particular "selected" microorganism.
For example, if a certain microbe is resistant to aparticular antibiotic (e.g., novobiocin), then
that antibiotic can be added to the medium in order to prevent other organisms, which are
not resistant, from growing.
Selective medium contain particular ingredients that inhibit the growth of certain microbes.
An example of a selective medium is MacConkey agar. It contains bile salts and crystal
violet, which interfere with the growth of many gram-positive bacteria and favor the growth of
gram-negative bacteria. A selective medium is a medium that allows the selection of one or
more types of microorganisms. These microorganisms will be the only ones able to grow on
or in the medium while all the others will be inhibited. Selectivity is achieved in several ways.
MacConkey agar is a selective and differentiating agar that only grows gram-negative
bacterial species; it can further differentiate the gram-negative organisms based on their
lactose metabolism.

Q 10:- An incubator is a device used to grow and maintain microbiological cultures or cell
cultures. The incubator maintains optimal temperature, humidity and other conditions such
as the CO2 and oxygen content of the atmosphere inside.
incubator, an insulated enclosure in which temperature, humidity, and other environmental
conditions can be regulated at levels optimal for growth, hatching, or reproduction. There are
three principal kinds of incubators: poultry incubators, infant incubators, and bacteriological
incubators.
incubator depends on the principle of thermo-electricity. The incubator has a thermostat
which maintains a constant temperature by creating a thermal gradient. When any conductor
is subjected to a thermal gradient, it generates voltage called as thermo-electric effect. There
are two types of incubators in relation to airflow: circulated air incubators and still-air
incubators. Circulated air incubators, also known as forced air incubators, have built-in fans
that continually circulate air to maintain sufficient oxygen and keep the temperature even.
An incubator is a device used to grow and maintain microbiological cultures or cell cultures.
The incubator maintains optimal temperature, humidity and other conditions such as the
CO2 and oxygen content of the atmosphere inside. An incubator is designed to provide a
safe, controlled space for infants to live while their vital organs develop. Unlike a simple
bassinet, an incubator provides an environment that can be adjusted to provide the ideal
temperature as well as the perfect amount of oxygen, humidity, and light.

Q 11:- Virology is the scientific discipline concerned with the study of the biology of viruses
and viral diseases, including the distribution, biochemistry, physiology, molecular biology,
ecology, evolution and clinical aspects of viruses. xamples include the invention of Influenza
model by Donald E. Ingber group, the invention of Ebola virus disease model by Alireza
Mashaghi group, and the invention of viral hepatitis model by Marcus Dorner group. The
organ chip approach will likely replace animal models for human virology.
Firstly, the increasing importance of virology is clearly linked to the fact that we know more
and more viruses, understand their links to certain diseases better and that epidemiology
 looks at certain viral infections in new ways: all of a sudden we recognise viruses where we
did not see them before.
Immunofluorescence or immunoperoxidase assays are commonly used to detect whether a
virus is present in a tissue sample. These tests are based on the principle that if the tissue is
infected with a virus, an antibody specific to that virus will be able to bind to it.
If it's a viral illness, typically symptoms are shorter lasting and classically the symptoms
include fever, chills, sore throat, nasal congestion, runny nose, cough, and a lot of times you
can have some body aches. A lot of times the symptoms last for maybe three days to a week
and then slowly get better over time.

1. Q12:- a toxin or other foreign substance which induces an immune response in the body,
especially the production of antibodies.
Listen to pronunciation. (AN-tih-jen) 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. In general, antigens are composed
of proteins, peptides, and polysaccharides. Any portion of bacteria or viruses, such as
surface protein, coat, capsule, toxins, and cell wall, can serve as antigens.
There are three main types of antigen

The three broad ways to define antigen include exogenous (foreign to the host immune
system), endogenous (produced by intracellular bacteria and virus replicating inside a host
cell), and autoantigens (produced by the host).
Listen to pronunciation. (AN-tee-BAH-dee) A protein made by plasma cells (a type of white
blood cell) in response to an antigen (a substance that causes the body to make a specific
immune response). Each antibody can bind to only one specific antigen. What are
Antibodies? Antibody (Ab) is also known as an immunoglobulin(Ig). These are large, Y-
shaped blood proteins produced by plasma cells. They bind to foreign particles and invade
them. Antibodies are proteins produced by the immune system in response to infection. They
are an important part of the body's defence system as they work to destroy disease-causing
organisms (such as viruses or bacteria) and block them from infecting human cells.

Q 13:- What happens during a CRP test? A health care professional will take a blood sample
from a vein in your arm, using a small needle. After the needle is inserted, a small amount of
blood will be collected into a test tube or vial. You may feel a little sting when the needle
goes in or out.
How is hs-CRP tested? CRP is measured with a simple blood test, called a high-sensitivity
CRP (hs-CRP), that can be done in the lab using a blood test. To save time, hs-CRP testing
can be done at the same time as cholesterol testing. However, it can also be done
separately.
A CRP test measures the levels of CRP in the blood. This can help detect inflammation due
to acute health conditions or monitor the disease severity in chronic conditions, including:
Bacterial infections, such as sepsis. Fungal infections. The C Reactive protein test has no
special requirement like fasting or anything. Only a blood sample is drawn out by means of
an injection from the veins of the forearm. People under medication should inform their
doctor before undergoing this test as certain medications can cause disturbances on test
efficacy.

Q14:- The main principle of widal test is that if homologous antibody is present in patients
serum, it will react with respective antigen in the reagent and gives visible clumping on the
test card and agglutination in the tube. The antigens used in the test are “H” and “O”
antigens of Salmonella Typhi and “H” antigen of S.
Widal test (slide method) helps to detect typhoid fever and paratyphoid fever which are
collectively known as enteric fever. Enteric fever is caused by the ingestion of food or water
which is contaminated by bacteria. This test is pIf the Widal test range is more than or equal
to 1:160 titre for antigen O and antigen H, then it indicates typhoid infection. For the
diagnosis of a Widal blood test, 1:20, 1:40, 1:60, 1:80, 1:160, and 1:200 titres need to be
included in the diagnosis to obtain the typhoid test report.erformed through the slide method.
Widal test recorded 81.5% sensitivity, 18.3% specificity, 10.1% positive predictive value and
89.7% negative predictive value. Stool culture showed 31.3% sensitivity, 91.5% specificity,
29% positive predictive value and 91.5% negative predictive value. The Widal test is one
method that may be used to help make a presumptive diagnosis of enteric fever, also known
as typhoid fever.

Q 16:- Life Cycle (intestinal hookworm infection):

The released rhabditiform larvae grow in the feces and/or the soil , and after 5 to 10 days
(and two molts) they become filariform (third-stage) larvae that are infective . These infective
larvae can survive 3 to 4 weeks in favorable environmental conditions.
An adult hookworm can live for 4 to 24 months in the hosts intestine. The only way to
determine if a dog or cat has hookworms is to check a fecal sample for intestinal parasites.
The life cycle of the hookworm has three stages: egg, larvae, and adult. The microscopic
eggs pass through the feces, where they hatch into larvae and contaminate the environment.
The larvae can survive for weeks or even months before infecting your dog.
Adult hookworms in the intestines lay eggs that are passed out of the body through
feces. Once in the environment, the eggs hatch into larvae. Larvae enter a new host's body
by penetrating the skin. Preferred hosts can also be infected by swallowing hookworm
larvae, such as by eating contaminated dirt.
Eggs are passed in the stool , and under favorable conditions (moisture, warmth, shade),
larvae hatch in 1 to 2 days. The released rhabditiform larvae grow in the feces and/or the soil
, and after 5 to 10 days (and two molts) they become filariform (third-stage) larvae that are
infective .