NAME : JOSPHAT NZUVA

REG : B140/20598/2020
UNIT NAME : PRINCIPLES OF PARASITOLOGY
UNIT CODE : SBC 210

MICROSCOPY

INTRODUCTION
Microscope is used to study small organisms that cannot be seen with our naked eye there are three types of
microscopes simple compound and dissecting microscopes. Dissecting microscope uses visible light to view
large and thick objects whereas compound microscope are used to view small specimen

OBJECTIVE
I. To study the care and uses of a microscope

Procedure
1. Position the microscope correctly and identify the essential part
2. Turn the lamp brightness to its lowest setting then switch on the microscope
3. Turn up brightness to about ¾ of its full power. Turn the rotating nose piece until the objective (lower)
clicks into position
4. Place the specimen slide, cover glass uppermost on the front of the stage. Gently holding back the spring
arm of the mechanical stage
5. While looking (not down the eye piece) turn the coarse focusing knob to bring the specimen closer to the
objective that is approximately 5mm from the objective.
6. Looking down through the eye piece slowly turn the coarse focusing knob in the opposite direction to
increase the distance between the specimen and the objective
7. The specimen will come into focus but blurred
8. Use the fine focusing knob to obtain a sharp image

Result
Discussion
Microscope is an instrument that produces enlarged images of small objects allowing the observer un-
exceedingly close view of minute structures at a scale convenient for examination and analysis. The magnifying
power of a microscope is an expression of the number of times the object being examined appears to be enlarged
and is a dimensionless ratio in the form X10
The familiar type of a microscope in which glass lenses are used to form the image.it can be simple consisting if
single lens or compound consisting of several optical components in line. The specimen is placed on a slide and
placed on the stage of the microscope
It is positioned over the light using the x-y knob which move the slide on the surface of the stage. Coarse
focusing knob is used for large scale movement. Condenser lens concentrates all the light rays on the specimen
to minimize illumination. Diaphragm controls the density of illuminations

Conclusion
Microscope functioned to help view the parts of microorganisms and cells that are invisible to the naked eyes in more
details, moreover, proper cleaning of lenses should be done as well to avoid accumulation of dust particles that will lead to
discomfort usage of the microscope when viewing the specimen.

References
1. Celis JE (ed) (1998) Cell Biology: A Laboratory Handbook, 2nd edn. San Diego: Academic Press.
2. Lacey AJ (ed) (1999) Light Microscopy in Biology: A Practical Approach, 2nd edn. Oxford: Oxford
University Press.
 ASSOCIATION IN ANIMALS

Introduction
Association is an intimate association between specifically distinct organism’s in which one organism live on or
with another (usually the larger organism) in order to obtain nourishment at the same time causes harm to the
host.
Different species of animals are associated with one another in different ways. I.e. Parasitism, phoresis.

Objectives
To provide for both physical and physiological needs of the animals
To maintain a healthy balance with the host immune system that allows parasites to survive and reproduce

Materials
i. Leaf with aphids
ii. Microscope (dissecting and compound)
iii. Plasmodium slide

Method/procedure
[Activity 1]: to observe a kale plant under a dissecting microscope

Procedure
i. Place the leaf on the stage of a dissecting microscope
ii. Make observation under the microscope

[Activity 2]: examine stained mammalian blood smear showing malaria parasites

Procedure
i. Place plasmodium slide on the stage of the microscope
ii. Observe under the lower objective lens and center the paramecium in the microscopic field
iii. Once you have located rotate to the high power objective lens and refocus to observe the paramecium in a
greater detail

Results
Plasmodium slide.

Discussion
1. Parasitism
It is an intimate association between specifically distanced organisms in which one partner usually
smaller lives on or within the other 0rganism usually larger in order to obtain substance.
One organism benefits (parasite) and the other organism (host) is harmed
Examples:
Tapeworms in intestines of some animals, get nutrients from the host and the host animal becomes sick
2. Mutualism
Refers to a relationship between organisms of different species that show an intimate association with
each other. Both of the participants acquire a nutritional advantage.
Example;
 An anemone and a down fish, the anemone provides shelter while the down fish provides the anemone
with nutrient in form of waste.
3. Phoresis
This is where the phorant (usually the smaller organism) is mechanically carried by the larger organism
(usually the host). There is no dependency involved.
Example;
Plasmodium parasite being carried by female anopheles mosquito.
It is an interaction between individuals of different species that result in positive effects 0n the interacting
population.
Example;
4. Commensalism
It’s a relationship where neither of the animals involved loses nor gains.
Example;
Bacteria it the mouth, it lives and does not cause any harm.
Conclusion
Different organisms of different species associate directly depending on the host

Reference
 Ames, B. N., J. McCann, and E. Yamasaki. 1975. Methods for detecting carcinogens and mutagens with
the Salmonella/mammalian microsome mutagenicity test. Mutat. Res. 31:347–364. [PubMed]
 Anderson, D., R. E. Willingham, G. H. Lampkin, and P. B. Medawar. 1961. The use of skin grafting to
distinguish between monozygotic and dizygotic twins in cattle. Heredity 5:379.
 Arnsten, A. F. T., and P. S. Goldman-Rakic. 1985. Alpha-adrenergic mechanisms in pre-frontal cortex
associated with cognitive decline in aged non-human primates. Science 230:1272–1276. [PubMed]

HELMINTHES
Introduction
Helmith are large macro parasites; adult can be seen with naked eyes
Many are intestinal worms that are soil transmitted and they affect the gastro-intestinal tract.
Other parasites e.g. schistosomes resides in blood vessels examples of this parasites are class cestodes
which comprise of the tapeworms trematodes(flukes)and nematodes which are the roundworms
Helminthes causes infections such as onchoncarsiasis, schistosomiasis and food borne trematodiasis

Objectives
a) To classify parasitic worms into major groups
b) Intensify primary causes of infection dues to this parasites

Materials
a) Liver flukes
b) Round worms
c) Tape worms
d) Dissecting microscope
Method
i. Place the sample of tapeworm liver fluke and round worm on the stae of the dissecting
microscope
ii. Make observations
Result
Discussion
Nematodes
Have a long, thin, unsegment tube-like bodies
They have longitudinal digestive tracts, psuedoceolom which acts as an hydrostatic skeleton to provide
rigidity

Cestodes
They are segmented flatworms with a scolex
They lack body cavity and are flattened to facilitate perfusion to all tissues
Have a long flat ribbon-like bodies with numerous segments

Trematodes/flukes
Have small flat leaf-like bodies with oral and ventral suckers
They do not have body cavity, they are also dorsal-ventrally flattened with bilateral symmetry

Diseases
Enchinococcosis caused by echinococcus tapeworm which affects the liver
Hook worm disease caused by ancylostoma duodenale . it causes anemia and mulnutrion

Conclusion
In conclusion study of helminthes provides information on the value of alternative control methods that can be used to
reduce worm burdens. ReguJar fuecal removal from camps and a single pre-winter treatment with moxidectin proved to
be cost-effective methods to control helminth parasites

Reference
1. Ash L, Orihel TC: Parasites: A Guide to Laboratory Procedures and Identification. American Society of
Clinical Pathologists, Chicago, 1987 .
2. Bogitsh BJ and Cheng TC: Human Parasitology. WB Saunders, Philadelphia, 1990 .
3. Castro GA: Trematodes: schistosomiasis. p 1710. In Kelly WN (ed): Textbook of Internal Medicine. JB
Lippincott, Philadelphia, 1989 .
4. Hunter GW, Swartzwelder JC, Clyde DF: A Manual of Tropical Medicine. 5th Ed. WB Saunders,
Philadelphia, 1976 .

Identification of parasites
 Introduction
Parasites infections are caused by intestinal helminthes and protozoans.faeces are the most frequent
specimens collected and examined for demonstration of parasites of the gastro intestinal tract. To
identify intestinal parasites, there are three techniques used; direct, floatation and segmentation
methods.

Objectives
I. To detect parasites when they are not found in a direct method and the symptoms, which suggest
intestinal parasite infection.
II. Increase the probability of isolating eggs which are few in number.
III. To evaluate the success of treatment.

Materials
i. Methylated blue glycerol solution.
ii. Formalin.
iii. Lugol’s iodine.
iv. Glass slide.
v. Cover slips.
vi. Physiological saline.
vii. Beaker.
viii. Microscope.
ix. Fecal matter.
x. Test tube.
xi. Applicator stick.
xii. Collecting tubes.
xiii. Surgical gauge.
xiv. Zinc sulphate solution.
xv. Centrifuge tube.
xvi. Ether.

Activity 1. Direct method

Procedure
I. Emulsify a small quantity of fresh feces in a few drops o physiological saline.
II. Mount the specimen on a clean glass slide and place a cover slip on top of it.
III. You may add iodine on the specimen for staing.
IV. Examine under the microscope using the 10X objective.

Results
 Activity 2. Floatation method
Procedure
I. Fill a test-tube (without a lip but with a completely smooth rim.) about one quarter full with zinc
sulphate solution.
II. Add approximately 1g (or 2ml if fluid) of feces into the tube.
III. Using a stick, emulsify the specimen in the solution.
IV. Fill the tube with the zinc sulphate solution and mix well.
V. Strain the mixture through gauze or sieve to remove large faecal particles.
VI. Return the suspension into the tube.
VII. Stand the tube in a completely vertical position in a rack.
VIII. Using a plastic bulb pipette add more zinc sulphate solution to fill glass tube to the brim.
IX. Carefully place a clean (grease free) coverslip on top of the tube. Avoid trapping an air bubbles.
X. Leave undisturbed for 30 – 45 minutes to give time for the eggs and cysts to float on the surface
of the mixture.
XI. Carefully lift the coverslip from the tube by a strait upward pull.
XII. Mount the coverslip facing downwards on a clean glass slide in a drop of lugol’s iodine solution.
XIII. Examine under the microscope using the 10X objective .use the 40X objective to examine cysts
and eggs.

Results
 Activity 3 sedimentation method
Procedure
I. Using a stick, emulsify approximately 1g (pea size) of faeces in about 4ml of formalin contained
in a tube.
II. Add a further 4ml formalin and mix well.
III. Stain emulsion through gauze or sieve into a beaker.
IV. Transfer the suspension into a centrifuge tube.
V. Add 3 – 4 ml of ether to the suspension.
VI. Shake vigorously with thumb covering the centrifuge tube opening.
VII. Centrifuge at 750 – 1000 (approximately 3000 rmp) for 1 minute.
VIII. Using a stick, loosen the layer of foecal debris from the side of the tube and invert the tube to
discard the supernatants (ether, faecal, and formalin )
IX. Holding the tube in an upright position add a drop of iodine into the pellets deposited at the
bottom of the centrifuge tube.
X. Stir.
XI. Transfer all the contents in the centrifuge tube into a glass slide.
XII. Carefully place a coverslip on top of the preparation without tapping air bubbles.
XIII. Examine the preparation under the microscope at 100X.examine small cysts and eggs at 400X.

Results
Discussion
In sedimentation parasites are sedimented by gravity or centrifugal force. One example sedimentation
method is the formal ether concentration method. This method is the most frequent used technique
because it concentrates a wide range of parasites with minimum damage to their morphology.
Floatation are techniques with which parasites are concentrated by being floated in solution of specific
density, that is, solutions that are denser than the parasite being concentrated. Unlike the formal ether
sedimentation technique, a single floatation technique cannot be used to concentrate a wide range of
parasites due to differences in the densities of the parasites and the damage that can be caused by
floatation fluids to some parasites. Example of the floatation method include zinc sulphate and saturated
Nacl floatation techniques
Conclusion
Fecal sedimentation, the majority of trematode eggs are too large and heavy to float reliably in the
floatation fluids normally used for nematode egg, they however sink rapidly to the bottom of the fecal or
water suspensions.
Centrifugal floatation effectively concentrates fertilized eggs of ascaris specie, trichuris species, hook
worm, hymenolepis spiecies and enterobius vermicularis.

References
a) Sakamoto Y., ishguro M, kitagawa G.akaike information criterion statistics
boston:D. reidel publishing company 1986
b) Wang R, LiJ, chen Y, zhang L, xiao L. widespread occurrence of crypyosporidium
infections in patients with HIV/AIDS epidemiology, clinical feature diagnosis and
therapy. Acta trop. 2018; 187:256-263. [Pub med][google scholar]