LABORATORY ACTIVITY #3

Angelica M. Revil July 22, 2020

BSN-101

Laboratory Work 3. Differential staining technique: Gram’s Staining

A. Make a summary and step-by-step discussion of the procedures on making a Bacterial

Smear and Gram Staining:

---> Summary and Step-by-step discussion of the procedure on making a Bacterial Smear

What is a Bacterial Smear?

-Is a thin layer of bacteria placed on a slide for staining. Preparing the smear requires
attention to a number of details that help prevent contamination of the culture and ensure
safety to the preparer. Each step in this procedure has an important reason, and each should be
followed carefully as a prerequisite to successful work.
-The purpose of making a smear is to fix the bacteria onto the slide and to prevent the
sample from being lost during a staining procedure. A smear can be prepared from a solid or
broth medium. Below are some guidelines for preparing a smear for a Gram-stain. of liquid
bacterial growth in the center of a clean slide.

Equipments and materials:

 Buns and burner
 Gloves
 Goggles
 Slides
 Bacteria
 Inoculating loop
 Striker
 Water
 Marker
Procedures on making a Bacterial Smear:
 Begin the exercise by disinfecting The laboratory desk with the available
disinfectant as directed by the instructor. Prepare a clean sheet of paper towel or
absorbent material to be used if there is a spill.
 Wash the glass slide with soap, rinse and dry thoroughly. Use the marker to mark
off areas of the slide where smears will be placed.
 Obtain cultures of the bacteria, and place the tubes in a suitable rack on the bench.
Sterilize the inoculating loop.
 Light the Bunsen burner. The smear is now ready to be prepared.
 Firmly hold the loop as you would hold a pencil,flame for a few seconds or until it
glows red hot. Allow the loop to cool several seconds.
 Take the bacterial culture in the opposite hand, remove the cap.

 Place the tip of the tube in the flame for few seconds. This will burn off any dust or
lint and kill any airborne organisms that might happen to fall into the tube.

 Insert the loop into the tube, and touch it very gently to the main portion of the
slant.

Be careful not to dig into the medium or scrape it. Remove the loop and reflame the tip
of the tube briefly twice,

then replace the cap,

and return the culture to the rack on the desk.

 Mix the loopful of bacteria with the loopful of water on the side.

 And swirl the liquid out of the dime. For broth cultures, simply swirl the loopful of
bacteria on the marked slide area.
 Complete procedure by reflamming the loop. You now have a bacteriual smear.

 Prepare the smears in the remaining spaces with different bacteria as directed by
the instructor.
 Air -dry the slide
 Heat-fix twice the smears by quickly passing the slide through the bunsen burner.
 Done.
---> Summary and Step-by-step discussion of the procedure on making a Gram Staining

What is 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.

-The purpose of gram staining is to help doctors to prescribed specific antibiotic for that
specific infection and identify the bacteria type.

Equipment

Bunsen burner, alcohol-cleaned microscope slide, water

Reagents

Crystal violet, Gram's iodine solution, acetone/ethanol 0.1% basic fuchsin solution

Procedures
1. Prepare a Slide Smear:

A. Transfer a drop of the suspended culture to be examined on a slide with an

inoculation loop. If the culture is to be taken from a Petri dish or a slant culture tube,
first add a drop or a few loopful of water on the slide and aseptically transfer a minute
amount of a colony from the Petri dish. Note that only a very small amount of culture
is needed; a visual detection of the culture on an inoculation loop already indicates
that too much is taken.

If staining a clinical specimen, smear a very thin layer onto the slide, using a wooden
stick. Do not use a cotton swab, if at all possible, as the cotton fibers may appear as
artefacts. The smear should be thin enough to dry completely within a few seconds.
Stain does not penetrate thickly applied specimens, making interpretation very
difficult.

B. Spread the culture with an inoculation loop to an even thin film over a circle of 1.5
cm in diameter, approximately the size of a dime. Thus, a typical slide can
simultaneously accommodate 3 to 4 small smears if more than one culture is to be
examined.

C. Air-dry the culture and fix it or over a gentle flame, while moving the slide in a
circular fashion to avoid localized overheating. The applied heat helps the cell
adhesion on the glass slide to make possible the subsequent rinsing of the smear with
water without a significant loss of the culture. Heat can also be applied to facilitate
drying the the smear. However, ring patterns can form if heating is not uniform, e.g.
taking the slide in and out of the flame.

2. Gram Staining:
 A. Add crystal violet stain over the fixed culture. Let stand for 10 to 60 seconds; for
thinly prepared slides, it is usually acceptable to pour the stain on and off
immediately. Pour off the stain and gently rinse the excess stain with a stream of
water from a faucet or a plastic water bottle. Note that the objective of this step is to
wash off the stain, not the fixed culture.

B. Add the iodine solution on the smear, enough to cover the fixed culture. Let stand
for 10 to 60 seconds. Pour off the iodine solution and rinse the slide with running
water. Shake off the excess water from the surface.

C. Add a few drops of decolorizer so the solution trickles down the slide. Rinse it off
with water after 5 seconds. The exact time to stop is when the solvent is no longer
colored as it flows over the slide. Further delay will cause excess decolorization in the
gram-positive cells, and the purpose of staining will be defeated.

D. Counterstain with basic fuchsin solution for 40 to 60 seconds. Wash off the
solution with water. Blot with bibulous paper to remove the excess water.
Alternatively, the slide may shaken to remove most of the water and air-dried.

3. Quality control:

It is a simple matter to prepare a control slide by breadking a clean wooden applicator

stick and picking a small amount of material from the interproximal space of one's
teeth. This should be smeared into a drop of clean tap water on a clean glass slide.
The slide may be stained as above. This material will consistently display a few
neutrophils and a mixture of Gram (+) and (-) organisms. Neutrophil nuclei should be
pink.

3. Examine the finished slide under a microscope.

A caveat in the examination of the Gram smears is the distortion in morphology that
can be caused by antimicrobial therapy. This is especially likely to occur in urine
speciments. Filamentous and pleomorphic forms may be observed among the Gram
(-) rod species. Gram reaction of the organism may also change after antimicrobial
therapy, Gram (+) bacterial may become gram variable. Look at areas that are one
cell thick only; observation of thick areas will give variable and often incorrect
results. White blood cells and macrophages should stain Gram-negative, whereas
sqamous epithelial cells are Gram-positive.

What happened to the slide?

- if Gram + - then the iodine will complex with the crystal violet forming a crystal when
dries out, it sticks within the wall and remain with a purple cell when decolorized.

-if Gram - = Lipid layer keep the iodine from getting in there and forming the complex
so its gonna wash off and counterstain with the safranin(a red stain) it makes cells pink.
Answer the following questions:

1. If your organisms were all blue, what is the Gram reaction?

- Gram’s method involves staining the sample cells dark blue,

decolorizing those cells with a thin cell wall by rinsing the sample, then
counterstaining with a red dye. The cells with a thick cell wall appear
blue (gram positive) as crystal violet is retained within the cells, and so
the red dye cannot be seen. 
2. If your organisms were all pink, what is the Gram reaction?
-A Gram negative bacteria should give a pink stain. This is becaue
it does not retain the crystal violet because the peptidoglycan layer is in
the periplasm. It’s Gram negative because lipid layer keep the iodine
from getting in there and forming the complex so its gonna wash off
and counterstain with the safranin a red stain which makes thhe cell
pink.
3. If your organisms were a mixture of blue and pink, what is
the Gram reaction? Why? How can that be proven?
-Initially, both Gram-positive AND Gram-negative cells are stained
by the Primary Stain, the Crystal Violet.  In the second step of the
procedure, Gram's Iodine is added to the smear as a mordant to
complex with the crystal violet and forms an insoluble complex in
Gram-positive cells.  At this point, the cell types will both appear
purple.  The dye-mordant complex will not be removed from
Gram-positive bacteria but is leached from Gram-negative cells
during the alcohol or acetone  (95% ethyl alcohol) in
the Decolorization step.  Decolorization is the most important step! 
Decolorization serves as a protein-dehydrating reagent and a lipid
solvent.  The alcohol increases the porosity of the cell wall by
dissolving the lipids in the outer layers of the bacterium.  The
crystal violet complex can then be more easily removed from the
thinner area and less cross-linked area of the peptidoglycan layer. 
After decolorization, Gram-positive cells still remain purple, but
Gram-negative cells are colorless.  In the final step, a counterstain,
safranin or carbol fuschin, is added to counterstain the Gram-
negative cells.

4. What would be the reason(s) for not finding any organisms

on the slide?
-
There are many reason this could have happened. The first
is that the researcher may not have been able to scrape any
of the actual cultures off the agar. Another could be that
the smear was not properly heat fixed to the slide and may
 have been washed away with the dye. Another reason is
that there are organisms on the slide but the researcher has
a difficult time focusing the microscope.
5. Give five examples of Gram positive bacteria and their
corresponding diseases.

Examples of high G+C gram-positive bacteria that are human pathogens include:
 Mycobacterium tuberculosis, which causes tuberculosis
  M. leprae, which causes leprosy (Hansen’s disease)
 Corynebacterium diphtheriae, which causes diphtheria.
 Clostridia spp. are low G+C gram-positive bacteria that are generally obligate
anaerobes and can form endospores. Pathogens in this genus
include C. perfringens (gas gangrene), C. tetani (tetanus), and C.
botulinum (botulism).
 Lactobacillales include the genera Enterococcus, Lactobacillus, Leuconostoc,
and Streptococcus. Streptococcus is responsible for many human diseases,
including pharyngitis (strep throat), scarlet fever, rheumatic fever,
glomerulonephritis, pneumonia, and other respiratory infections.

6. Give five examples of Gram negative bacteria and their

corresponding diseases.
Common gram-negative bacteria and the infections and diseases they
cause include:

 Escherichia coli (E. coli)—food poisoning, urinary tract infections,

gastroenteritis, and newborn meningitis
 Pseudomonas aeruginosa—lung and urinary tract infections
 Klebsiella—meningitis, and lung, urinary tract, and bloodstream
infections
 Acinetobacter baumannii—several types of infections in wounded
soldiers
 Neisseria gonorrhoeae— gonorrhea , a sexually transmitted disease
 Enterobacteriaceae—urinary tract, lung, and bloodstream
infections, and food poisoning (includes carbapenem-resistant
Enterobacteriaceae, which are very resistant to antibiotics)
 LABORATORY ACTIVITY #4

Angelica M. Revil

BSN-101

ACID-FAST

Acid-fast stain is a differential stain used to identify acid-fast organisms such as

members of the genus Mycobacterium. Acid-fast organisms are characterized by wax-
like, nearly impermeable cell walls; they contain mycolic acid and large amounts of
fatty acids, waxes, and complex lipids. This type of cell wall is resistant to most
compounds, therefore acid-fast organisms require a special staining technique. The
primary stain used in acid-fast staining, carbol fuchsin, is lipid-soluble and contains
phenol, which helps the stain penetrate the cell wall. This is further assisted by the
addition of heat in the form of heat (steam). Steam helps to loosen up the waxy layer
and promotes entry of the primary stain inside the cell. The smear is then rinsed with
a very strong decolorizer, which strips the stain from all non-acid-fast cells but does
not permeate the cell wall of acid-fast organisms. The decolorized non-acid-fast cells
then take up the counterstain, which in our case is methylene blue.

PREPARASYON PARA SA MICROSCOPYO NGA SLIDE:

 Hinloe ang mga slide gamit ang Kimpwe og alkohol para mawala ang mga
fingerprints

 Pag drawing og duha ka bilog gamit imo marker ubos sa imo slide

 Gamit ang inoculation loop, butangi og duha ka gagmay na patak sa tubig kada
bilog

 Gamit ang limpyo na pa-maagi, kuha e ra ng gamay ra na bakteria sa culture tube,

siguraduha lang na tagpabaga nimo ang culture tube bag o ka musulod og
pagsulod nimo dakan preperado

 Mansahan ang bakteria sa pagtulo sa tubig sa imong slide.

 Pasagdi na mumala ang slide, pamalha sa hangin.

 E init nga pag-ayo ang slide sa nagdagan pinaagi sa siga sa basya ka tulo sa ka-
pat ka beses lamang uban ang smear tupad na pamaagi. Ayaw bagahi ang kilid
nga naay bakteria.

 Tapos, pasagdi na mo mala para andam na mag mantsa.

UNSAON PAGMANTSA:

 Tabuni ang mga smears gamit ang taulya sa papel sulod sa utlanan sa slide.
 Ibutang ang slide sa ibabaw sa usa ka beaker na nagapaagay nga tubig. Ayaw
pasagdi na mag dry ang tubig sa beaker.

 Pagbaha sa papel na talya nga adunay carbonfuchsin. Og pakulo e ang slide ang
slide tulo sa ka lima ka munito

 Kuha a ang tualya na papel pagkatapos tapos ilabog na sa basurahan

 Hugasi ang slide gamit ang tubig para mahinlo og mawala ang mga salin salin sa
tualya nga papel tapos og taptapi na pagpamaya

 But-ngi dayon og methylene blue na magtagal ng 1.5 minuto.

 Hugasi napud og tubig dayon taptapi na pagpamaya

 Pag-blot sa kalumo gamit ang bibulous nga papel

 pamalha nga e dry ang utlanan sa slide bag-o ibutang sa entablado sa

mikroskopyo tapos tan -awa dayon gamit ang oil immersion lens.

ANSWER THE FOLLOWING QUESTIONS:

1. What is the primary and counter stain for acid fast stain?
-The primary stain used in acid-fast staining, carbol fuchsin, is lipid-
soluble and contains phenol, which helps the stain penetrate the cell
wall. This is further assisted by the addition of heat in the form of heat
(steam). Steam helps to loosen up the waxy layer and promotes entry of
the primary stain inside the cell. The smear is then rinsed with a very
strong decolorizer, which strips the stain from all non-acid-fast cells but
does not permeate the cell wall of acid-fast organisms. The decolorized
non-acid-fast cells then take up the counterstain, which in our case is
methylene blue. While the Counter stain is the smear is stained with
counterstain, methylene blue. Only decolorized cells absorb the counter stain
and take its color and appears blue while acid-fast cells retain the red color
2. Describe the structure that is responsible for allowing certain species to be
acid fast. What species should be positive for the acid fast stain?
-Mycobacterium smegmatis is positive for acid fast stain; The structure that
allows certain species to be acid fast is the waxy mycolic acid in the cell wall.

3. What is the decolorizer used for acid fast stain?

-Th decolorizer for acid fast stain is the metheylene blue,The smear is then
rinsed with a very strong decolorizer, which strips the stain from all non-acid-
fast cells but does not permeate the cell wall of acid-fast organisms. The
decolorized non-acid-fast cells then take up the counterstain, which in our case
is methylene blue.
4. Give two examples of acid fast negative bacteria and the corresponding
diseases.

-klebsiella pneumonia- Cause of pneumonia

 -Staphylococcus auerus- Common cause of skin infections including
abscesses, respiratory infections such as sinusitis, and food poisoning.

5. Give two examples of acid fast positive bacteria and the corresponding
diseases.
-Mycobacterium tuberculosis causes the
respiratory disease tuberculosis

-Mycobacterium leprae causes the disfiguring disease leprosy.

Both are treatable with long-term multi-drug therapies.

Laboratory Work #5
Endosphore Staining
Angelica M. Revil
BSN-101

“endospore staining, and reading the procedure, make a summary and step-by-
step discussion of the procedures on making Endosphore Staining..

-The endospore stain is a differential stain used to visualize

bacterial endospores. Endospores are formed by a few genera of bacteria,
such as Bacillus . By forming spores, bacteria can survive in hostile
conditions. Spores are resistant to heat, dessication, chemicals, and
radiation. Endospore staining is a differential stain that aims at detecting,
identifying and differentiating an endospore from the vegetative cell (an
underdeveloped endospore). The principle of the role is to detect the
presence or absence of the endospore, but some procedures have
modified the technique by increasing the concentrations of the dyes,
increasing the duration of heat fixing, application of ultraviolet radiation.
With the improved technology in microscopy, some use phase-contrast
microscopy which is fast and it produces more detailed morphologies of
the bacterial endospore.

Procedure:
 Pag andam sa mga smear ng mga microorganismo para pag test sa
mga presensya ng mga endosphores sa hinlo na slide sa microskopyo
tapos e pamala sa hangin or aird dry.
 E pag ayo sa kainit sa lkalayo ang smear.
 Butangi nan gamay na piraso ng musuhop na papel sa ibabaw sa
smear tapos ibutang ang slide sa wire gauze gamit ang ring stand.
 Paso a ang mga slide hangtod sa mag sugod na kini pag alisbo( pwede

pinaagi sa pagbutang sa slide sa usa ka staining rack na gibutang

ubabaw sa bunsen burner o sa gakulo na water bath.
 Pagnga ang kalayo o e paso napud ang slide kung kinahanglanon para
mapadayon ang pagpa init o singaw sa slide sa 3-5 munotos. Kung
ang papel nagsugod nag ka uga butangi og malachite green para
magpabilin nga umog, ayaw pud tawon pasobrahi
 Pagkatapos nan 5 minutos palihog kuhaa na ang slide, ignayi gajud
pagkuha sa rack gamit ang clothespin.
 Kuha a ang blotting paper og pasagdi na mabugnaw ang slide sa 2
minutos.
 Banwasi ang slide og ayo gamit ang tubig para ma wala ang malachite
green sa kada slide nan mikoskopyo na slide
 Mantsaha butngi color ang slide ng Safranin mga 2 minutos.
 Banwasi ang tagkakilid nan slide para ma remove ang ikaduhang
mantsa og e pamala dayon sa hangin o e air dry.

- Endospore staining uses two stains to differentiate endospores from the rest

of the cell.

1. Schaeffer Fulton Stain- used Malachite Green dye and

safranin
2. Dorner method of endospore staining –uses Carbolfuchsin
stain, acid alcohol, and Nigrosin solution)

1. The Schaeffer-Fulton method (the most commonly used endospore-

staining technique) uses heat to push the primary stain (malachite green) into
the endospore. Washing with water decolorizes the cell, but the endospore
retains the green stain. The cell is then counterstained pink with safranin. The
resulting image reveals the shape and location of endospores, if they are
present. The green endospores will appear either within the pink vegetative
cells or as separate from the pink cells altogether. If no endospores are present,
then only the pink vegetative cells will be visible.
 2. Dorner's Method staining technique- It is an old method of endospore
staining, which makes the use of three reagents, namely primary stain,
decolourizer and counterstain. To make the process faster The
endospore stain is a differential stain which selectively stains bacterial
endospores.

The results are Vegetative cells appear colorless, while endospores are red.

Laboratory Work 6. Special staining: Negative staining

Angelica M. Revil
BSN-101

CAPSULE STAINING
Summary and step-by-step discussion of the procedures on performing capsule
staining in the vernacular or Surigaonon.

-Capsule Staining- is a type of differential stain which uses acidic and basic dyes

to stain background & bacterial cells respectively so that presence of capsule is easily
visualized. Capsule is synthesized in the cytoplasm and secreted to the outside of the
cell where it surrounds the bacterium.
-Certain bacteria and yeasts have a protective outer structure called a capsule. Since
the presence of a capsule is directly related to a microbe’s virulence (its ability to
cause disease), the ability to determine whether cells in a sample have capsules is an
important diagnostic tool. Capsules do not absorb most basic dyes, therefore, a
negative staining technique (staining around the cells) is typically used for capsule
staining. The dye stains the background but does not penetrate the capsules, which
appear like halos around the borders of the cell. The specimen does not need to be
heat-fixed prior to negative staining.

Materials and reagents required

 Test bacteria: 36-48 hour culture of capsulated bacteria e.g. Klebsiella
pneumoniae growing on a slant of EMB Agar or culture of other capsulated
bacteria and non-capsulated bacteria [Note: Growing Klebsiella pneumoniae in
milk-based media (e.g. Skim milk) increase its capsule size, making it easier to
visualize.]
 Stain solutions: Depending on the type of method used (Crystal violet, India ink,
Nigrosin, Copper Sulfate, Basic carbol fuschin solution, Methylene blue solution,
etc).
  Microscopic slides
 Inoculating loop
 Microscope with 100x objective lens (oil immersion)
 Immersion oil
 Gas burner
 Tissue paper

Capsule Stain procedure

A. India Ink Method

 Butangi og isa ka patak ng Idia ink sa limpyo na microscope slide. Diha sa kilid
sa nagbagtok nga ngilit.
 Gamit ang Flamed loop og limpyo na pamaagi, butngi og tulo ka lopho nga
gisulayang bakterium gikan sa broth culture. Og dugangan nimo og bakteria
gikan sa culture plate siguraduha nga walay dagko nga organismo pero tinguhaa
nga malikayan ang pagkaylap.
 Ibutang ang tumoy sa usa ka limpyo nga slide sa mikroskopyo sa usa ka anggulo
hangtod sa katapusan sa slide nga adunay micoorganismo. Mikaylap ang
pagbutang sa usa ka pilmahi. Kini buhaton pinaagi sa pagkontra sa tinulo sa
biyolet na crystal nga adunay limpyo nga slide sa mikroskopyo og gigamit ang
aksyon nga capillary sa dye/slide para pagkatag sa kristal nga violet sa tibuok nga
smear.
 Itugot ang pilmahi nga mo maya sa hangin mga 5-7 minutos
 Ikiling ang slide og waswasi mga 20 porsyento nga copper sulfate solusyon.
Ayaw waswasig tubig kay mawala ang mga capsula gikan sa selula.
 Pamalha ang slide sa hangin nga pamaagi or air dry bisag dali lang nga minuto
 Og pagkatapos obserbahi ang slide sulod sa oil immersyon.

Resulta:
Lantawa ang mga purpura nga selula nga gilibutan sa usa ka tin-aw o mahuyang nga
asul nga halo sa transparent nga background. Ang halo mao ang kapsula.

B. Anthony’s stain method

 Ibutang ang usa ka tinulo nga kristal nga violet sa usa ka limpyo nga slide sa

mikroskopyo, nga kasikbit sa frosted edge

 Using a flamed loop and sterile technique, add three loopful of test
bacterium (any capsulated bacteria such as Klebsiella pneumoniae,
Streptococcus pneumoniae) from broth culture. If you are adding bacteria from a
 culture plate make sure that there are no large clumps of the organism, but try to
avoid spreading the drop.

Ibutang ang tumoy sa lain nga limpyo nga mikroskopyo nga slide sa usa ka
anggulo ngadto sa katapusan sa slide nga adunay sulod nga organismo. Ipakaylap
ang paghulog sa usa ka pilmahi.
 Ibutang ang tumoy sa usa ka limpyo nga slide sa mikroskopyo sa usa ka anggulo
hangtod sa katapusan sa slide nga adunay micoorganismo. Mikaylap ang
pagbutang sa usa ka pilmahi. Kini buhaton pinaagi sa pagkontra sa tinulo sa
biyolet na crystal nga adunay limpyo nga slide sa mikroskopyo og gigamit ang
aksyon nga capillary sa dye/slide para pagkatag sa kristal nga violet sa tibuok nga
smear.
 Itugot ang pilmahi nga mo maya sa hangin mga 5-7 minutos
 Ikiling ang slide og waswasi mga 20 porsyento nga copper sulfate solusyon.
Ayaw waswasig tubig kay mawala ang mga capsula gikan sa selula.
 Pamalha ang slide sa hangin nga pamaagi or air dry bisag dali lang nga minuto
 Og pagkatapos obserbahi ang slide sulod sa oil immersyon.

QUESTIONS:

1. Enumerate the different dyes/stains used in capsule staining and mention

their purpose

-Bacterial capsules are non-ionic, so neither acidic nor basic stains will adhere to their

surfaces. Therefore, the best way to visualize them is to stain the background using an
acidic stain and to stain the cell itself using a basic stain. We use India ink and
Gram crystal violet method with two different kinds of stains primary stain and
the counterstain.

1. India Ink - India ink is often used as the negative stain.

However, some of the other stains that can also be used include:

 Congo red
 Nigrosin
 Eosin

This is a negative staining technique that is essentially used to identify the presence of
capsules. Because of its acidic nature, India ink (or Congo red, nigrosin) stains the
background dark.

2. crystal violet is used for number of reasons including:

 To act as a fixative
 Increase penetration power
 Stain the cells (being a basic dye)
 Decrease pH of smear
 When viewed under the microscope, the background will be dark as a result of India
ink, bacteria cells will be purple having taken the crystal violet dye while the capsule
will be clear against a dark background given that it takes no stain.

2. Why is capsule stain considered differential stain?

-Because it's going to differentiate bacteria that has capsule and that dose not
have.
3. Explain how a capsule increases the pathogenicity of bacteria with
capsules.
-The capsule is considered a virulence factor because it enhances the ability
of bacteria to cause disease (e.g. prevents phagocytosis). The capsule can
protect cells from engulfment by eukaryotic cells, such as macrophages.
A capsule-specific antibody may be required for phagocytosis to occur.
Capsules also contain water which protects the bacteria against desiccation.
They also exclude bacterial viruses and most hydrophobic toxic materials such
as detergents. Immunity to one capsule type does not result in immunity to the
other types. Capsules also help cells adhere to surfaces. As a group where the
capsule is present they are known as polysaccharide encapsulated bacteria or
encapsulated bacteria. A bacterial capsule has a semi-ridged border that
follows the contour of the cell. The capsule excludes India Ink when dyed. A
slime layer is a non-ridged matrix that is easily deformed and is not able to
exclude India Ink. Biofilms are composed of many cells and their outer
barriers. The primary functions of both capsules and slime layers are for
protection and adhesion.
4. Give two examples of capsulated bacteria and their corresponding
diseases.
1. Salmonella typhi-are bacteria that infect the intestinal tract and the blood. The
disease is referred to as typhoid fever. S.
...
Typhi may cause:

 Constipation, more common than diarrhea.

 High fever.
 Headache.
 Fatigue.
 Loss of appetite.
 Dizziness.
 Cough.
 A rash on the trunk.

 2. Streptococcus pneumoniae and Streptococcus pyogenes- infections may

take the form of pharyngitis, scarlet fever (rash), impetigo, cellulitis, or erysipelas.
Invasive infections can result in necrotizing fasciitis, myositis and streptococcal toxic
shock syndrome

5. Give two examples of bacteria without capsules and corresponding

diseases.
1. Haemophilus influenzae type b -causes pneumonia,
septicaemia, meningitis, epiglottitis, septic arthritis, cellulitis, otitis
media, and purulent pericarditis, as well as less common
invasive infections such as endocarditis, osteomyelitis, and peritonitis.

2.Pneumococcal disease is caused by bacteria called Streptococcus

pneumoniae (pneumococcus). People with pneumococcal disease can
spread the bacteria to others when they cough or sneeze.

Pneumococcus bacteria can cause infections in many parts of the body,

including

 Lungs (pneumonia) 
 Ears (otitis)
 Sinuses (sinusitis)
 Brain and spinal cord tissue (meningitis)
 Blood (bacteremia)