Clinical Parasitology Module: For The Use of University of Baguio School of Natural Sciences Students Only

Clinical
Parasitology Module
For the Use of University of Baguio School of
Natural Sciences students only
Compiled and Prepared by

Jaleh V. Gacayan, RMT, LPT, MPH
Jessa Felix, RMT
Erlinda P. Sanchez, RMT, MPA,LIB
A Self-regulated Learning Module 2
INTRODUCTION
Clinical parasitology is an area of biology concerned specifically to the
study of important parasites which causes diseases to man. It covers the
classifications of parasites, symptoms, diseases caused by the parasites,
lifecycle, transmission, and treatment. The diseases caused by these parasites
constitute major human health problems throughout the world.
Parasitology is an essential component of clinical laboratory medicine.

The results obtained through proper specimen examination of parasites will
provide crucial information with regards to the diagnosis and treatment of
human disease. Tracking the epidemiology of such organisms as well as
establishing preventive mechanisms may be accomplished with the assistance
of this information.
In spite of the many advances in technology that has been developed in

recent years, the conventional technique of manually processing and
examining the samples both macroscopically and microscopically still occurs in
select clinical settings. It is essential that well-educated and extremely trained
individuals perform these procedures as well as read and interpret the results.
Thus, the goal of this module is to provide the needed information for students
preparing for a career in medical laboratory science. The module is designed to
help the learners in both the lecture and laboratory components related with
clinical parasitology. Students using this module will have the opportunity to
advance their skills necessary to become proficient laboratory scientists.
COURSE TITLE AND DESCRIPTION

This module is a learning package for the CLPRST 1-Clinical Parasitology
offered as a professional course in Bachelor of Science in Medical Laboratory
Science. It comprises of three (3) units lecture and one (1) unit laboratory.
This course deals with the study of human parasites which are of medical
importance especially those commonly found in the Philippines. Emphasis is
given on the epidemiology, pathogenecity, distribution, life cycle, and
laboratory identification of each parasite. Preventive measures against infection
and control are also given emphasis.
ABOUT THE MODULE

This module was designed to adopt the blended learning approach in
teaching Clinical Parasitology to the BSMLS students via online and distance
learning.
Learning Engagement1 of the module deals with the overview of what
Clinical Parasitology is. It includes an introduction which covers the terminologies
commonly used in the course, the different ways of specimen processing to
isolate a parasite, Techniques Used in the Identification of Parasites as well as
prevention and control. It also covers the morphology, pathophysiology, life
cycle, specimens used for the identification, diagnostic features, prevention and
control of Nematodes (Roundworms).
Learning Engagement 2 of the module is about the Morphology,
Pathophysiology, Life cycle, Specimens used for identification, Diagnostic
features, Prevention and Control of Cestodes (Tapeworms). It also deals with
the Morphology, Pathophysiology, Life cycle, Specimens used for identification,
Diagnostic features, Prevention and Control of Trematodes (Flukes).
Learning Engagement 3 covers the Morphology, Pathophysiology, Life
cycle, Specimens used for identification, Diagnostic features, Prevention and
Control of Protozoa. It also deals with the characteristics and role of

ectoparasites affecting man.
COURSE REQUIREMENTS
The pre-requisite assigned for the course is Human Anatomy and
Physiology.
Student’s attendance in the online learning sessions is a basic requirement

in which university policy on tardiness and absences are applied. Various
assessment tools are employed in the course to gauge the student’s level of
understanding and comprehension within the duration of the online learning
sessions throughout the term.
It is hoped that by the end of the course, students have instilled the
necessary and essential skills in clinical parasitology that would equip them in
their practice of the profession.
Requirements of the Course

1. Regular Attendance to classes: You must attend online classes and live quizzes
regularly by logging in to our scheduled online activities. Online lectures will
be done through Google meet and/or facebook live. Assessments shall be
given through Quizziz, Pear Deck, Canvas and/or Google forms. For offline
students, your attendance will be monitored through your responses to text
information and through timely correspondence.
2. Submission of required activities: All required activities (assignments, research
work, laboratory illustrations) should be submitted on or before the given
deadline. Deadlines will be posted by the teacher in the google classroom and
messenger group chat. It will also be texted to offline students. For online
students, requirements must be submitted to the teacher’s email address which
will be provided during the class orientation. For offline students, requirements
must be submitted via mail or express courier (e.g. LBC, JRS) addressed to:
Instructor’s name, School of Natural Sciences, University of Baguio, Baguio City.
3. Seventy percent (70%) passing score in all required activities: Quizzes, exams,
assignments, research work, laboratory illustrations.
Computation of grades:
▪ The Course Grade is obtained by combining the lecture and laboratory
grades (50%:50%) for the subject.
▪ Laboratory grade shall be computed as 30% enhancement activities
(illustrations; research work; case study; experiments – when possible) plus
70% class standing (quizzes and exams).
▪ The cumulative system of computing grades shall be followed. Grades
computed for midterms and finals are considered tentative. The final
midterm grade is calculated by getting 1/3 of the first grading grade plus 2/3
of the tentative midterm grade and the final grade is computed by getting
1/3 of the midterm grade plus 2/3 of the tentative final grade.
4. Study/Learning Guidelines:
a. Manage your time properly. As students of higher education (College), you
are expected to be more responsible in paying attention to course
schedules, requirements, and deadlines. Schedule how you will accomplish
all the requirements in all your enrolled courses (reading the modules, reading
on research/ enhancement questions, doing assignments and laboratory
illustrations) and focus your attention when doing your tasks.
b. Observe proper conduct. Despite this online mode of learning, you must still
maintain appropriate behavior at all times. All standards of student conduct
outlined in the University of Baguio Student Handbook remain in full effect
during this time of distance learning. Be honest in answering your quizzes and
exams. Work independently when accomplishing tasks and assignments.
c. Stay motivated. Your future depends on what you do today. Maintain a

positive attitude towards learning and enjoy a fun-learning environment
despite the current circumstances.
d. Maintain a performance of high standard. Give your best in accomplishing all
the assigned tasks. Do not be complacent with just a 70% passing cut-off
score. Remember that this is a board subject, and the best preparation for the
board/licensure examination should be during these formative years. The
board review is but supplementary to the knowledge you have already
learned during your Med Tech education.
e. Communicate properly. Promptly respond to notifications by regularly visiting

our google classroom and messenger group chat. If you have confusions or
queries in any part of this module, I am here to guide you through. Send your
academic concerns using the same online platforms. For offline students, text
messages and mobile calls are welcome during scheduled hours of the day
and week. Be guided by this schedule when communicating:
▪ Respect private hours. I do not always open my laptop/email/messenger

24/7. Send your queries and/or concerns during regular office hours. For
concerns that need immediate attention, send through mobile text.
▪ Be patient. Messages received between 8 AM to 8 PM will be responded

to within the same day. Messages received after 8 PM will be answered
starting 8 AM the next day.
▪ Before calling my mobile number, text first for permission for I might be
giving an online lecture or in a meeting or on private moment at that
very instance.
▪ Saturdays and Sundays are for my family and home chores. I shall
respond to queries/messages received during these days within the first
office hour of Monday.
f. Show mutual support. Support one another. Let us all be responsible and
supportive in making this new learning process more effective.
g. Live lecture/Video conferencing guidelines:

g.1Be punctual. Live lectures/Video conferences will be scheduled during the
official class period/time of this course. Log in to the platform at least 5-10
minutes before the class period. Prepare your learning materials such as
this module, pens, papers, etc. Attendance will be checked during the
lecture/video conference.
g.2Maintain professionalism.
- Wear appropriate clothing and set your gadget in an appropriate

area. You may be asked to turn on your video/camera at any time
during the lecture.
- Log in using your UB gmail account. Unidentified names like nicknames,

phone models, etc. will not be allowed in the video conference.
- Mute your microphone as soon as you log in to the platform to avoid

any excess background noise. Unmute your microphone when
instructed to do so.
- Be courteous. Do not interrupt your teacher or a classmate who is
speaking. You may type your question in the Chat area, or use the
“raise hand” feature if available, and wait until you are allowed to
speak.
- Respect privacy. Do not take a screenshot, picture, snapshott, etc. of

your teacher or fellow students, nor make any unnecessary audio or
video recordings.
g.3Remain focused and engaged. Do not be distracted by your gadget.
Keep your videoconference platform open and do not navigate other
tabs or webpages unless directed by your teacher.
Endorsed by:
Teresa N. Villanueva, RMT, MACT

Dean, School of Natural Sciences
STUDY SCHEDULE
WEEK TOPIC ACTIVITY
1 Section 1 Introduction to Online Lecture
Parasitology Laboratory : Experiment 1 & 2
2 Continue Section 1 Introduction to Online Lecture, Assessment & Quiz

Parasitology Laboratory : Experiment 3 and 4
3 Section 2: Nematose Intro Online Lecture: Assessment &

Section 2A Quiz Laboratory: Experiment 5
Adenophorea
4 Section 2B: Phasmidea Online Lecture: Assessment & Quiz Laboratory:
Experiment 6 and 7
5 Section 2B: Phasmidea Blood and Quiz Laboratory: Experiment 8
Tissue Nematodes and 9
Online Lecture: Assessment &
6 FIRST GRADING EXAM Coverage Section 1 and 2
7 Section 3: Cestodes - Introduction Laboratory: Experiment 10
Online Lecture: Assessment & Quiz
8 Section 3: Cestodes Online Lecture: Assessment & Quiz Laboratory:
PRACTICALS
9 Section 4: Trematodes - Quiz Laboratory: Experiment 11
Introduction
Online Lecture: Assessment &
10 Section 4A: Dioecious Flukes Online Lecture: Assessment & Quiz Laboratory:
Experiment 12
11 Section 4B: Monoecious Flukes Online Lecture: Assessment & Quiz Laboratory:
PRACTICALS
12 MIDTERM Coverage Lesson 1-10 13 Section 5A: Sarcodina (Amoeba) Online
Lecture: Assessment & Quiz Laboratory: Experiment 13
14 Section 5B: Mastigophora Online Lecture: Assessment &
(Flagellates) Quiz Laboratory: PRACTCALS
15 Section 5C: Mastigophora Online Lecture: Assessment &
(Hemoflagellates) Quiz Laboratory: Experiment 14
16 Section 5D: Ciliophora (Ciliates) Online Lecture: Assessment & Quiz
Laboratory: Experiment 15
17 Section 5E: Apicomplexa (Sporozoa) Online Lecture: Assessment & Quiz
Laboratory: Experiment 16
18 Section 6: Ectoparasites Online Lecture: Assessment & Quiz Laboratory:
EXAMINATION
19 FINALS Coverage Lesson 1-17
Table of Contents
Section 1: Introduction to Parasitology.............................................................................................11
Experiment no. 1: Instrument and Glasswares ............................................................................ 37
Experiment no. 2: Physiology of Stool Formation and Specimen Collection ............................... 40
Experiment no. 3: Routine Stool Examination (Physical and Chemical Phase)............................. 43
Experiment no. 4: Routine Stool Examination (Microscopic) ...................................................... 46
Section 2: Nematodes .....................................................................................................................52
Section 2A: Adenophorea................................................................................................................... 56
Experiment no. 5: Phylum Nematoda (Aphasmidea) ................................................................. 67 Section
2B: Phasmidea ....................................................................................................................... 72 Experiment
no. 6: Phylum Nematoda (Phasmidea) .................................................................. 103 Experiment no. 7:
Phylum Nematoda (Secernentia; Phasmidea) ............................................. 109 Experiment no. 8: Phylum
Nematoda (Blood and Tissue Nematodes) ..................................... 117 Experiment no. 9: Phylum
Nematoda (Other Members) ......................................................... 122 Section 3: Cestodes
.......................................................................................................................125 Experiment no. 10:
Phylum Platyhelminthes (Class Cestoda) .................................................. 145 Section 4: Trematodes
...................................................................................................................150 Section 4A: Dioecious
Flukes ........................................................................................................... 154 Experiment no. 11:
Phylum Platyhelminthes (Class Trematoda – Dioecious flukes).................. 163 Section 4B: Monoecious
Flukes ....................................................................................................... 167 Experiment no. 12: Phylum
Platyhelminthes (Class Trematoda – Monoecious flukes).............. 199 Section 5: Protozoa
.......................................................................................................................204 Section 5A: Sarcodina
(Amoeba) ...................................................................................................... 209 Experiment no. 13:
Phylum Sarcomastigophora (Subphylum Sarcodina) .................................. 233 Section 5B: Mastigophora
(Flagellates) ............................................................................................ 237 Section 5C: Mastigophora
(Hemoflagellates) ................................................................................... 246 Experiment no. 14: Phylum
Sarcomastigophora (Subphylum Mastigophora) ........................... 256 Section 5D: Ciliophora (Ciliates)
....................................................................................................... 262 Experiment no. 15: Phylum
Ciliophora....................................................................................... 265 Section 5E: Apicomplexa (Sporozoa)
................................................................................................ 268 Experiment no. 16: Phylum
Apicomplexa (Class Sporozoa) ....................................................... 284 Section 6: Ectoparasites
......................................................................................................................... 289
LEARNING ENGAGEMENT 1 CLINICAL PARASITOLOGY: OVERVIEW
INTRODUCTION:
Parasitology is the area of biology concerned with the phenomenon of
dependence of one living organism to another. Medical Parasitology is
concerned primarily with the animal parasites of humans and their medical
significance, as well as their importance in human communities. It includes the
study of three major groups of animals: parasitic protozoa, parasitic helminths
(worms), and the arthropods that directly cause disease or act as vectors of
different pathogens. A parasite is a pathogen that simultaneously injures and
derives sustenance from its host. Some organisms called parasites are actually
commensals, in that they neither benefit nor harm their host (for example,
Entamoeba coli). Although parasitology had its origins in the zoologic sciences,
it is today an interdisciplinary field, greatly influenced by microbiology,
immunology, biochemistry, and other life sciences. The basic biology of the
pathogens, their host-parasite relationships and descriptions of the basic
properties of the pathogens, the pathogenesis of the diseases they cause, host
defenses, and epidemiology are highlighted in this module.
SECTION 1: INTRODUCTION TO PARASITOLOGY
COURSE LEARNING OUTCOMES:
At the end of this session, the students must have:
1. used the basic terminologies and theories related to Parasitology 2.

enumerated the precautions and guidelines to be considered in proper
specimen collection
3. appreciated the significance of laboratory waste management, prevention and
control of parasitic infections, and the maintenance of a safe laboratory
environment
4. established guidelines on acceptability of stool samples in the laboratory 5.
interpreted and decided on the negativity or positivity of the sample studied and
follow the correct manner of reporting
LESSON PROPER
I. TERMINOLOGIES
PRE-ACTIVITY: Find the meaning of the following terminologies and indicate the
reference used for each.
1. Parasitology 4. Commensalism 5. Mutualism
2. Parasitism 6. Parasite
3. Symbiosis a) Ectoparasite
15. Coprozoic/spurious parasite 16.
b) Endoparasite
Host
7. Facultative parasite 8. Obligate
parasite 9. Erratic parasite 10. 17. Final/definitive host
Incidental parasite 18. Intermediate host
11. Temporary parasite 19. Accidental host
12. Permanent parasite 20. Paratenic host
13. Pathogenic parasite 14. 21. Dead-end host
Pseudoparasite 22. Reservoir host
A. PARASITE-HOST RELATIONSHIPS
23. Parasitic infection
24. Parasitic infestation
27. Oviparous female parasite 28.
25. Parasitic disease
Parthenogenic female parasite
26. Larviparous/viviparous
29. Protandry
female parasite
II. PARASITIC INFECTION
Organisms may develop unique relationships due to their habitual and long
associations with one another. These relationships are very important to their survival.
Symbiosis is the living together of unlike organisms. It may involve protection or other
advantages to one or both partners. Different forms of symbiosis may be distinguished
on the basis of whether or not the association is detrimental to one of the two
partners. Commensalism is a symbiotic relationship in which two species live together
and one species benefits from the relationship without harming or benefiting the other.
Mutualism is a symbiosis in which two organisms mutually benefit from each other.
Parasitism is a symbiotic relationship where one organism, the parasite, live in or on
another, depending on the latter for its survival and usually, at the expense of the host.
Table 1-1 lists the terms associated with parasite-host relationships, along with their
definitions.
A Self-regulated
Learning Module 13
There are several types of parasites that may be members of a parasite-host

relationship. An organism may be an obligatory parasite or a facultative parasite. It
may be an endoparasite or an ectoparasite. In the same manner, a number of
different hosts may be part of this parasite host relationship. These include accidental
or incidental hosts, definitive hosts, intermediate hosts, reservoir hosts, transport hosts,
and carriers.
When a parasite infects a host, symbiosis results. The primary function of the host
is to carry on the parasite’s life cycle. This newly formed relationship may develop into
commensalism, mutualism, or parasitism. Some of these associations exist as
commensal under certain circumstances and pathogenic under others. Parasites
have an amazing capability to adapt to their host surroundings. In addition to a
number of morphologic adaptations, parasites
are capable of protecting themselves from the host’s immune system. Parasites alter
their antigenic makeup so that the host will not recognize the modified parasites as
foreign, and thus the initiation of an immune response does not
occur.
B. EXPOSURE and INFECTION
Majority of animal parasites are pathogens which are harmful and which
frequently cause mechanical injury to their hosts. A carrier harbors a particular
pathogen without manifesting any signs and symptoms.
Exposure is the process of inoculating and infective agent, while

infection connotes the establishment of the infective agent in the host.
The incubation period is the period between infection and evidence of symptoms. It
is sometimes referred to as the clinical incubation period. The pre-patent period, also
known as the biologic incubation period, is the period between infection
or acquisition of the parasite and evidence of demonstration of infection.
Autoinfection results when an infected individual becomes his own direct source of
infection. On the other hand, superinfection or hyper infection happens when the
already infected individual is further infected with the same species leading to
massive infection with the parasite.
C. SOURCES OF INFECTION
There are various sources of parasitic infections. The most common sources are
contaminated soil and water. Lack of sanitary toilets and the use of night soil or
human excreta as fertilizer allow the eggs to get in contact with the soil. Another
possible source of infection is water and food, which contain the infective stage of the
parasite. Consumption of undercooked or raw fresh water fish can result in several
intestinal and liver fluke infections. Arthropods can also transmit infection. Mosquitoes
are vectors of malaria and filaria parasites. Other animals, whether wild or
domesticated, may also harbor the parasite. Other sources include another person,
his beddings and clothing, the immediate environment he has contaminated, or even
one’s self.
D. MODES OF TRANSMISSION
An infectious agent may be transmitted from its natural reservoir to a
susceptible host in different ways. There are different classifications for modes of
transmission:
1. Direct – contact w/ an infected person or animal, directly from the source
to the susceptible host without involving an intermediate object
a) Droplet spread
b) Sexual intercourse
c) Kissing
d) Holding hands
e) Transplacental / Vertical - mother to fetus
2. Indirect
a) Ingestion of contaminated food & drink
b) Contact w/ contaminated soil
c) Bite of an infected arthropod (vector)
d) Through fomites
Since the most common source of parasitic infection is contaminated food and
water, the most likely portal of entry is the mouth. Majority of infections among
cestodes, trematodes, and intestinal protozoans are foodborne: Taenia solium, Taenia
saginata, and Diphyllobothrium latum from eating food harboring the infective larval
stages; Entamoeba histolytica and Giradia lamblia from drinking water contaminated
with cysts; and Clonorchis, Opistorchis and Haplorchis through ingesting raw or
improperly cooked freshwater fish containing the parasitic larvae.
Skin penetration is another route of transmission. Hookworms and Strongyloides enter
via exposure of skin to soil, while Schistosoma species enter skin via water.
Arthropods also serve as vectors and transmit parasites through their bites. Examples
are agents of malaria, filariasis, leishmaniasis and trypanosomiasis. Another way of
acquiring infection is through congenital transmission.
Toxoplasma gondii trophozoites can cross the placental barrier during pregnancy. In
transmammary infection with Ancylostoma and Strongyloides, the parasites may be
transmitted through the mother’s milk.
Other ways of acquiring the infection include inhalation of airborne eggs of
Enterobius, and sexual intercourse as in the case of Trichomonas vaginalis.
E. LIFE CYCLE
Although parasitic life cycles range from simple to complex, they all have three
common components—a mode of transmission, a morphologic form that invades
humans, known as the infective stage, and one (or more) forms that
can be detected via laboratory retrieval methods, known as the diagnostic stage.
Some parasites require only a definitive host, whereas others also
require one or more intermediate hosts. A parasitic life cycle consists of two common
phases (Fig. 1). One phase involves the route a parasite follows when in or on the
human body. This information provides an understanding of
the symptomatology and pathology of the parasite. Insights about the best the
method of diagnosis and selection of appropriate antiparasitic medication may also
be determined. The other phase, the route a parasite follows independently of the
human body, provides crucial information pertinent to epidemiology, prevention,
and control.

Figure 1. Generic Parasite life cycle
F. DISEASE PROCESSES and SYMPTOMS

A parasitic disease may affect the entire body or any of its parts. The major
body areas associated with such processes include the following: (1) the
gastrointestinal (GI) and urogenital (UG) tracts; (2) blood and tissue; (3) liver, lung, and
other major organs; and (4) miscellaneous locations, such as cerebrospinal fluid (CSF),
eye, skin, and extremities.
A wide variety of representative symptoms, may occur when a parasite
infects a human host. Some persons remain asymptomatic, whereas other parasites
produce severe symptoms and may result in death. The most commonly observed
symptoms include diarrhea, fever, chills, abdominal pain, and abdominal cramping.
Other symptoms, such as elephantiasis (an enlargement of areas such as the breast,
leg, and scrotum caused by a parasite’s presence), anemia, vitamin deficiency,
bowel obstruction, edema, enlargement of major organs, skin lesions, and blindness,
may develop.
G. PREVENTION and CONTROL

Prevention and control measures may be taken against every parasite infective
to humans. Preventive measures designed to break the transmission
cycle are crucial for successful parasite eradication. Examples of such measures
include the following: education programs, use of insecticides and other chemicals,
protective clothing, protective netting, proper water treatment, good personal
hygiene, proper sanitation practices, proper handling and preparation of food, and
avoidance of unprotected sexual relations. The vast capital expenditures required to
accomplish these measures are not available in many endemic countries in the world.
The problem of eradicating parasites is an
ongoing process and is a key goal of international health groups such as the World
Health Organization (WHO)
III. LABORATORY METHODS FOR DIAGNOSIS
A. SPECIMEN FOR DIAGNOSIS

The following are the specimens needed to properly diagnose the presence of
parasites inside the human body.
1. Stool- for intestinal protozoans, nematodes and helminthes
2. Urine- for the recovery of Trichomonas vaginalis and Schistosoma
haematobium
Collection: mid-stream catch
3. Sputum- Paragonimus westermani, larvae of nematodes
• Must be digested using 4-5% sodium hydroxide
4. Blood- for malarial parasites, filarial worms, Leishmania and Trypanosoma
5. Cerebrospinal fluid- Acanthamoeba species

• Collection: lumbar tap
6. Tissue aspirates:
a) Liver aspirate- hydatid cyst and liver amoebic abscess
b) Duodenal aspirate- Giardiasis and Strongyloidiasis infection
• Collection: endoscopy
• Duodenal drainage or “String test”: duodenal contents collected for
Giardia and Strongylodes
• Sigmoidoscopy: Schistosomiasis, Amoebiasis, Balantidiasis and Shigellosis
(Large intestines)
c) Broncho-alveolar lavage – Paragonimus westermani
7. Orifice swab
a) Vaginal swab- Trichomonas vaginalis
b) Perianal swab – Enterobius vermicularis and Taenia
8. Tissue Biopsy
a) Muscle- Trichinella spiralis
b) Rectal – granulomas secondary to Schistosomiasis
B. PROCESSING OF BLOOD SAMPLE
There are parasites where the method of isolation and identification is through
processing of blood.
❖ Blood Films
1. Fresh water smears- for diagnosis of Trypanosomes and microfilaria
2. Thin Dry smears – for the study of the morphology of the parasites and the blood
cells
3. Thick Dry smears – used for malaria survey among patients with chronic infections
or who are undergoing anti-malaria therapy.
❖ Stains
1. Giemsa – most preferred
Composition: Stock solution 1 ml
Buffered water (pH 7.0 – 7.2) 49 ml
Staining time: 30 minutes
- Too dark : acidic pH
- Too red: alkaline pH

2. Rapid stains:
a) Wright’s stain – It is used to stain blood smears in the detection of blood
parasites. The stain distinguishes easily between blood cells and became
widely used for performing differential white blood cell counts, which are
routinely ordered when infections are expected. The stain contains a fixative,
methanol, and the stain in one solution. Thin films of blood are fixed with
methanol to preserve the red cell morphology so that the relationship
between parasites to the red cells can be seen clearly.
✓ Fix with 1-2 drops of methanol
✓ Cover the film with 10% Giemsa stain: 5 minutes
✓ wash with distilled water, drain, dry and examine
b) Leishmann stain – It is a mixture of Methylene blue, and Eosin dye, prepared in
Alcohol medium and diluted with buffer or distilled water during staining
procedure. Leishman stain is a differential stain that is used to variably stain the
various components of the cells and it can be used to study the adherence of
pathogenic bacteria to the human cells. It differentially stains the human and
bacterial cells and appeared as purple and pink colored bodies respectively.
The Leishman stain is one of the best stains for routine blood stain to stain the
Peripheral blood smear for the examinations of blood film under the
microscope and is satisfactory for malaria and other blood parasites
✓ Add 7-8 drops of the stain: 1-2 minutes
✓ Add 12-15 drops of buffered distilled water
✓ Mix thoroughly, let stand (4-8 minutes)
✓ Rinse, drain, dry and examine
c) Field’s stain - It is a histological method for staining of blood smears. It is used for
staining thick blood films in order to discover malarial parasites. Field's stain
consists of two parts - Field's stain A is methylene blue and Azure 1 dissolved in
phosphate buffer solution; Field's stain B is Eosin Y in buffer solution.
d) Acridine orange - Acridine Orange Stain is used as a fluorescent staining agent
to detect the presence of malaria parasite in blood cultures and other bodily
fluids. Acridine orange is a fluorochrome dye that can interchalate into nucleic
acid.
e) Jaswant Singh Battacharya (JSB) Stain for thick and thin films - standard
method: laboratories under the National Malaria Eradication Programme in
India
3. Permanent stains & Other stains
a) Iron Hematoxylin Stain - used for most of the original morphological descriptions
of intestinal protozoa found in humans.
b) MIF Fixative Stain (Merthiolate iodine formaldehyde) - diagnosis of
Trichomonas
c) Chlorazol Black E - An acid dye, used as a fat and general tissue stain, and to
stain protozoa in fecal smears or in tissues.
d) Modified Kohn’s - modification of the chlorazol black E staining technique
e) Wheatley Trichrome - all-purpose (amoebae, flagellates)
f) Methenamine Silver - cyst
g) Fluorescent Staining- Microsporidium
PARASITEMIA
Thick Blood Film:
✓ Infected erythrocytes are counted in relation to a predetermined number of

WBCs
✓ Standard: average of 8000/µl
✓ All parasite species and forms (sexual and asexual) are counted
Formula:
# of parasites/µl = # of parasites x 8000

# of WBCs counted
• If parasites are ≥10, 200 leukocytes are counted:
# of parasites/µl =# of parasites counted x 40
• If parasites are <10, 500 WBCs should be counted:
# of parasites/µl = # of parasites counted x 16
✓ Earle and Perez method

• # of asexual parasites/ 5µl
• thick film is used
• used only in research studies
Thin Blood Film:
✓ % of parasitemia: P. falciparum
✓ # of infected red cells in 1000 RBCs
✓ smear is scanned carefully, one 'row' at a time
✓ total number of red cells and the number of parasitized red cells are tabulated
separately
✓ If 1000 red cells are counted:
• divide the number of parasitized red cells by 10 to get the percentage •
less than 1000 red cells counted
• # of parasitized cells X 100
# of RBCs
"plus system"
✓ less precise
✓ variation in the thickness of the film
✓ results in variation in parasite count
+ = 1–10 per 100 thick fields
++ = 11-100 per 100 thick fields
+++ = 1–10 per thick field
++++ = >10 per thick field
C. COLLECTION OF FECAL SAMPLE

Proper collection of fecal sample is essential for the isolation and proper
diagnosis of infection. The following should be considered:
1. Container: sterile, disposable, wide mouth with tight-fitting lid, transparent

(for physical examination of the specimen)
2. Avoid contamination with urine, water and soil

3. Label
4. Handle carefully because it is a potential source of infection
unsuitable samples from patients receiving:
a. Barium
b. Oil
c. Bismuth
d. Kaolin
e. Antibiotics
TYPES of FECAL SPECIMEN
1. Liquid – either diarrheic or saline-purged

2. Soft
3. Formed
PRESERVATION
a) Physical:
1. Room temperature
2. Refrigeration
b) Chemical
1. Polyvinyl Alcohol (PVA) – used for the preservation of stained fecal+
smears; preservation of trophozoites
2. 5-10% formalin
▪ For concentration techniques or direct fecal smear
▪ For cysts, helminthes and larvae
▪ Not sufficient for preparing permanent stained fecal smears
3. Merthiolate-Iodine-Formalin (MIF) or Thimerosal Iodine Formalin (TIF)
▪ Good for all stages
▪ For liquid stools
▪ For bulk feces, the solution is mixed in the proportion of 9.4 ml
MIF and 0.6 ml Lugol’s for a gram of fecal material
4. Schaudinn’s Fixative
▪ For fresh materials and those recovered from intestinal

mucosal linings
▪ For permanent staining
IMPORTANCE OF FECALYSIS
1. To detect the presence of intestinal parasites

2. For the detection of evidence of malfunction of some parts of the GIT, liver
and pancreas
3. For the detection of GIT bleeding
4. For the detection of excessive fats in stool (steatorrhea)
5. Used as a clue in medical and surgical diagnosis.
D. EXAMINATION OF FECAL SAMPLE

1. Physical Examination
❖ Form and Consistency:
Normal: soft to formed
Variations
a) Very soft and watery – seen in diarrhea and administration of saline
cathartic
b) Excessively hard and scybalous – constipation due to lack of mucus
c) Rice water stool – cholera
d) Pea soup stool – early typhoid
e) Flattened or ribbon like – syphilis, spastic colitis and obstruction at the lower
portion of the colon
f) Butter like- fibrocystic disease of the pancreas
g) Gaseous and fermentative – excessive carbohydrate fermentation
❖ Color
Normal: light brown to dark brown due to stercobilin
Variations
a) Yellow- due to administration of santonin and senna
b) Light clay or putty color – seen after ingestion of Barium meals
c) Reddish or bloody- bleeding in the lower GIT; undigested beets and
tomatoes
d) Dark red/chocolate brown- bleeding in the upper GIT

e) Black/tarry- associated with digestion of blood due to bleeding in the
upper GIT
f) Greenish – amoebiasis
❖ Odor
Normal: foul to offensive due to indole, skatole and butyric acid
Abnormal odors:
a) Putrid odor – found in ulcerative and malignant tumor of the lower bowel
b) Sour/rancid odor- indicates gas formation, fermentation of carbohydrate,
unabsorbed fatty acids
c) Extremely foul odor – usually in alkaline stools, putrefaction of undigested
protein.
2. Chemical Examination
Occult blood – blood which is not visible to the naked eye and can only be
detected by chemical means only
Laboratory Examination for Occult Blood:
a) Benzidine test
b) Guaiac;s test
c) Hematest
General Principle: The peroxidase-like activity of blood (due to peroxidase
contained in the heme portion of hemoglobin) decomposes H2O2 to produce
water and oxygen. The liberation of oxygen causes the oxidation of the
colorless chromogen into a colored compound (blue)
Patient Preparation: meat free diet for 3-5 days prior to the test
3. Microscopic Examination
The following structures may be seen microscopically:
a) Trophozoites and cysts of amoeba
b) Helminth eggs and larvae
c) RBC- due to hemorrhagic disorder, ulcers and contamination
d) Macrophages present in bacterial or parasitic infection
e) WBC- indicative of inflammation
f) Fungi
g) Plant cells, pollen grain and spores
h) Epithelial cells
i) Calcium oxalate and triple phosphate crystals
j) Bacteria
k) Plant fibers, root hairs and animal cells ( similar to helminth ova)
l) Charcot-layden crystals
TECHNIQUES
1. DIRECT FECAL SMEAR – simplest and most frequently used

a) 0.85% sodium chloride/NSS- for the observation of the motility of trophozoites
b) D’antonis solution/Lugol’s Iodine – for protozoan cysts and helminth ova 2. KATO
THICK SMEAR (KTS)
▪ Reagents:
Distilled water 100 ml
Glycerin 100 ml
3% malachite green 1 ml
▪ Use cellophane as cover slip
3. CONCENTRATION TECHNIQUES- used in the detection of a small number of
parasites which are not detected using DFS
a) Sedimentation Method – uses either specific gravity or centrifugation •
Formalin-Ether techniques – concentrates helminth eggs, larvae
and protozoan cysts
• Acid-Ether Sedimentation – used for formed stools; applicable for
helminth eggs and larvae
• Simple sedimentation – time-consuming
• Merthiolate-Iodine-Formalin Concentration
• Acid Sodium sulfate Tritan
• Knott Concentration Technique- for concentration of
microfilariae’ utilizes venous blood as specimen
b) Flotation method

✓ Allows the separation of helminth eggs, protozoan cysts and
larvae (1.05 – 1.15) using chemical solutions of higher specific
gravity (1.12-1.21)
✓ Eggs and cysts float to the surface of the solution while fecal
materials sink to the bottom
✓ Superior to sedimentation for concentrating cysts and eggs other
than operculated, schistosomal and infertile Ascaris eggs
• Zinc Sulfate centrifugal flotation technique
- valuable method for concentrating cysts and eggs
- specific gravity: 1.18 (hydrometer); 1:20
formalinized feces
• Sugar Flotation Technique/Sheather’s Sugar Flotation
- Cryptosporidium (rounded oocysts)
• Wilie’s Brine- uses sodium chloride solution
• Lane’s Direct Centrifugal Technique
• Magnesium Sulfate Techniques
4. STAINING METHODS
a) Temporary Staining
• D’Antonis
• 1% Isotonic Eosin Solution & 2% Brilliant Cresyl Blue
- Trophozoites- blue green
- Background- pink
• Eosine in saline
• Buffered Methylene Blue (ex. Nairs Methylene Blue)
b) Permanent Staining
• Gomori’s Trichrome Stain: The trichrome technique of Wheatley for
fecal specimens is a modification of Gomori's original staining
procedure for tissue. It is a rapid, simple procedure which produces
uniformly well stained smears of the intestinal protozoa, human cells,
yeast cells, and artifact material in about 45 min or less.
- Wheatley’s modification

- For intestinal protozoa
• Lawless’ Permanent Mount Stain: A rapid permanent-mount stain
technic for the diagnosis of the intestinal protozoa
- Rapid method of staining trophozoites and cysts
- Protozoa – blue to purplish color
• Modified Acid-Fast stain
- Cryptosporidium, Isospora and Cyclospora
- Two separate stains: Carbol fuchsin and methylene blue
- lacks specificity
- Use of the modified Ziehl-Neelsen stain for faecal smears has
already been established for coccidian protozoa, in particular,
oocysts of Cryptosporidium species, but it is also useful to confirm
the presence of oocysts of Isospora belli and Cyclospora
cayetanensis
• Iron Hematoxylin Stain
- used for most of the original morphological descriptions of
intestinal protozoa found in humans.
- On oil immersion power (1,000 x ), one can examine the
diagnostic features used to identify the protozoan parasite. Iron
hematoxylin staining methods can be used with either fresh, SAF
preserved, or PVA-preserved specimens.
• MIF Fixative stain ( Merthiolate iodine formaldehyde)
- diagnosis of Trichomonas
• Chlorazol Black E
- An acid dye, used as a fat and general tissue stain, and to stain
protozoa in fecal smears or in tissues.
• Modified Kohn’s
- modification of the chlorazol black E staining technique
5. SPECIAL RECOVERY METHODS
a) Cellulose Tape Preparation/Graham Scotch Tape Method
✓ Enterobius vermicularis and Taenia
✓ Collection: morning before the patient washes or defecates
✓ Commercial collection kits are available

b) Egg Count Technique – provides an estimate of worm burden
✓ Determines the degree of infection (light, moderate or heavy)
✓ For recovery of hookworms, Ascaris and Trichuris
✓ Methods:
1. Direct Smear Egg Count (Method by Beaver)
- 1.5 mg feces (667) = eggs per gram (epg)
- 2.0 mg feces (500) = epg
2. Dilution Egg Count – uses 0.1 N NaOH
- Stoll’s Egg Counting Technique
3. Thick Smear Egg Count – KTS for schistosomes
4. Dilution-Filtration Egg Count – Schistosomes
c) Egg Hatching Technique
✓ Egg hatch assay (EHA) is a laboratory tool used to determine a given
parasite's resistance to extant drug therapy.
✓ Fresh eggs are incubated from the parasite of interest and serial
dilutions of the drug of interest are applied. The percentage of eggs
that hatch or die is determined at each concentration and a drug
response curve may be plotted. The data can then be transformed
and analysed to give further statistics such as an ED50.
✓ This technique is labour intensive, expensive and can take some time,
however an egg hatch assay will give and accurate and reliable
result.
6. CULTURE METHODS
PURPOSES:
✓ For accurate detection of the organisms as a supplement to other
methods
✓ For obtaining a rich yield of organisms to be used as antigen in
immunologic diagnosis
✓ For in-vitro screening of drugs
✓ For investigating the physiology of organisms
✓ As a source for inoculating susceptible experimental animals
CULTURE METHODS for PROTOZOA

a) Balamuth’s Monophasic Medium
- All liquid medium commonly used for the isolation and maintenance
of Entamoeba histolytica and some other intestinal protozoa.
- It is satisfactory for Balantidium coli, although the quality and amount
of starch may need to be adjusted
- Use of egg yolk
b) Boeck & Drbohlav’s Diphasic as Modified by Dobell and Laidlaw
- Egg slant
- E. histolytica
- Concentrate from saline centrifugal sedimentation of feces rather
than the feces is recommended.
- Use of whole hen’s eggs
c) Cleveland & Collier’s
- This medium is highly specific for E. histolytica , which grows luxuriantly
on this medium. Other intestinal amoebae do not grow readily on this
medium. The technique of overlaying the medium with fresh sterile
horse serum-saline mixture, as reported by Cleveland and Collier, was
reported to be the best method of isolation of E. histolytica .
- Proteose peptone and infusion from liver provide amino acids and
other nitrogenous substances that support growth of E. histolytica .
- Sodium chloride maintains the osmotic balance of the medium and
sodium a-glycerophosphate acts as a phosphorous source.
d) Trussel and Johnson’s medium
✓ Isolation of Trichomonas vaginalis
e) Axenic Culture Method
- histolytica
- Axenic cultivation has been an essential component of numerous
immunological and biochemical studies
- Monophasic clear liquid for rapid growth of dense populations of
amebae
CULTURE METHOD for LEISHMANIA & TRYPANOSOMES
a) Novy, MacNeal and Nicole (NNN) Diphasic

- Best medium
- Use of sterile defibrinated rabbit blood

b) Cellular Media
- For Trypanosoma cruzi and Toxoplasma
IMMUNOLOGIC or SEROLOGIC TESTS – involves antigen-antibody reactions

a) Indirect Hemagglutination (IHA)
- PRINCIPLE: Patient serum and commercial sheep blood cells coated
with the appropriate antigen are required. The serum and cells are
combined and allowed to react. The test sample is then examined
for the presence of red blood cell agglutination
- USES: amoebiasis, Chaga’s disease, malaria, toxoplasmosis,
cystisercosis, hydatid cyst, filariasis, schistosomiasis
b) Flocculation or Agglutination
- PRINCIPLE: The patient serum is added to known cultured parasites.
This mixture is observed for the presence of parasite clumping.
- USES: Leishmaniasis, Chaga’s disease
c) Indirect Fluorescent Antibody (IFA)
- PRINCIPLE: Microscopically visible parasites fixed on a slide are
allowed to react with test serum. The slide is washed and treated with
anti-human globulin with fluorescein (sandwich method)
d) Gel Diffusion (Ouchterlony) and Countercurent Electrophoresis (CEP) -
PRINCIPLE: Patient serum and known antigens are placed in adjacent wells
located on the agar gel. The antigen and antibody are allowed
to migrate across the agar towards one anther. Specific patterns of
precipitate ( precipitin bands) result between the inoculated wells
and are then interpreted.
- USES: Detect amebiasis and fascioliasis infections
e) Enzyme -linked immunosorbent assay (ELISA)
- PRINCIPLE: Test serum is allowed to react with an antigen (coat the
surface of plastic well). Anti-human IgG antibody conjugated with
enzyme is added. If specific antibody was present in the test serum,
the anti-human IgG antibody labeled with an enzyme attaches to
the antigen-antibody complex. The addition of a suitable substrate

generates a visible color reaction.
- USES: T. gondii, T. vaginalis, leishmaniasis, malaria, schistosomiasis etc.
❖ Antibody Capture or Double Sandwich ELISA
✓ Modification of the standard ELISA for increased sensitivity and
specificity in detecting antibodies of the IgM class to
Toxoplasma
✓ Plastic well are coated with anti-human IgM antibodies
✓ After washing, the test serum is added followed by further
washing and addition of the test antigen
✓ The final reagent is an antigen-specific antibody to which the
color reagent has been conjugated
✓ The plates are read spectrophotometrically
✓ This method can be used for assay of parasite-specific IgE
antibodies by starting with anti-human IgE
f) Sabin-Feldman Dye Test
- The Sabin-Feldman Dye Test is based on the fact that live Toxoplasma
tachyzoites can actively take up methylene blue dye from the
culture medium, whereas parasites that are killed because of
complement-mediated lysis do not take up the dye and remain
colorless.
- Specifically designed for the diagnosis of Toxoplasmosis
g) Circumoval Precipitin Test (COPT)
- The circumoval precipitin test is a serological test used for diagnosis of
schistosomiasis japonica. Soluble egg antigens of Schistosoma
japonicum block the formation of the circumoval precipitin by serum
from infected humans. Consequently, circumoval precipitin inhibition
was used to monitor purification of the soluble egg antigens of S.
japonicum.
h) Radioimmunoassay (RIA)
- PRINCIPLE: Radioimmunoassay (RIA) is a very sensitive in vitro assay

technique used to measure concentrations of antigens by use of
antibodies. As such, it can be seen as the inverse of a radiobinding
assay, which quantifies an antibody by use of corresponding
antigens
i) Radioallergosorbent Test (RAST)
- The RAST is a radioimmunoassay test to detect specific IgE antibodies
to suspected or known allergens for the purpose of guiding a
diagnosis about allergy
- In this test, the parasite antigen is first bound to an inert sorbent or
matrix to which the test serum is allowed to react. After washing,
radio-labeled antibody (IgE or IgG) is added to the sorbent. After
another washing to remove excess labeled antibody, the remaining
radioactivity is a measure of antigen-specific IgG or IgE in the test
serum
j) Immunoblotting/Western Blot
- PRINCIPLE: CSF and serum may be used. By using electrophoresis
technique, the DNA containing portion of the patient sample is
transferred to a polyacrylamide gel. This process results in the
separation of protein antigens in the gel. A blotting technique is
employed to transport the proteins onto a paper.
- USES: giardiasis
k) Complement Fixation (CF)
- PRINCIPLE: Three ingredients is necessary to for performing the first
phase: patient serum, a commercial antigen source, known amount
of complement. The complement is “fixed” by the antibodies present
in the serum. In the 2nd phase, sensitized sheep’s rbc is added to the
mixture. The absence of free complement prevents lysis of these cells
( positive). If the patient sera does not contain Ab, complement is
free to lyse ( negative)
- USES: leishmaniasis, Chaga’s disease, pneumocystosis, paragonimiasis
7. ANIMAL INOCULATION TESTS
a) Leishmania
✓ Animal Used: hamster
✓ Specimen: aspirates from biopsy materials from cutaneous ulcers,
lymph nodes, spleen, liver
b) Trypanosoma
✓ Animal Used: Guinea pigs or white rats (Trypanosoma gambiense
and T. rhodesiense)
✓ White mice (Trypanosoma cruzi)
8. XENODIAGNOSIS
✓ An experimental bug (vector) is allowed to take a blood meal from the
patient
✓ The intestinal contents of the bug is assessed for the presence of
diagnostic stages of the parasite
SYNTHESIS
Over the years, parasites which were once considered commensal have evolved to
become human pathogens. During this time, tremendous knowledge was obtained of the
epidemiology, parasite-host relationships, life cycles, disease processes and symptoms,
treatment, and prevention and control of parasites. In addition, parasites are classified
based on their individual characteristics. Traditional as well as new methodologies for
parasite identification allow for accurate laboratory diagnosis.
Parasitology is an interesting and exciting field of the clinical laboratory sciences. The
continued development of high-tech, highly sensitive parasite test methodologies provides
the key to the future of parasitology. Because it is highly unlikely that parasites will totally be
eradicated in the near future, competent practitioners educated in the field of parasitology
are essential to ensure proper parasite identification.
ASSESSMENT
I. RESEARCH ACTIVITY
✓ In not more than 5 sentences, describe and indicate the purpose of the new
techniques in parasitology (20 points). Indicate the reference used. Use your own
words to explain.
1. DNA HYBRIDIZATION PROBE TESTING (DNA PROBE)
2. DNA RESTRICTION FRAGMENT LENGTH POLYMORPHISMS
3. FLOW CYTOMETRY
4. POLYMERASE CHAIN REACTION (PCR)
RUBRIC FOR GRADING:
II. QUIZ
✓ For offline and weak connectivity, it will be provided by the instructor in essay form ✓
For Online/Strong connectivity, it will be provided either through a worksheet, or quizziz or
google forms.
REFERENCES:
Belizario, V. Jr. (1998). Philippine Textbook of Medical Parasitology (1st edition). The
Publications Program.
Zeibig, A. (2013). Clinical Parasitology: A Practical Approach. (2nd ed.). Saunders Elsevier Inc.
Our Wright’s Stain (n.d). Dalyn biologicals (n.d)Retrieved from https://www. dalynn.com
/dyn/ck _assets/ files/ tech/SW80.pdf on July 3, 2020
Leishman staining- Principle, Preparation, Procedure & Result Interpretation (2018).

Paramedics World. Retrieved from https://paramedicsworld.com/hematology
practicals/leishman-staining-principle-procedure-interpretation/medical-paramedical
studynotes on July 3, 2020
CLINICAL PARASITOLOGY LABORATORY
LABORATORY GUIDELINES
❖ In case of any future face-to-face classroom/laboratory instructions. Please follow the
guidelines below
STUDENTS
1. Be in your complete white uniform (with nameplate and school ID). 2. Wear your
laboratory gowns at all times while inside the laboratory. 3. Incomplete/No uniform /No
Laboratory gown = No laboratory performance grade. 4. Put all cellular phones in the
silent mode. Answering of cell phones will be dealt with accordingly.
5. Wash hands thoroughly with soap and water before leaving the laboratory. 6. Long hair
should be clipped, fingers and hands should be kept away from the face and mouth.
ROOM MANAGEMENT & PROPER WASTE DISPPOSAL
1. Strictly no eating inside the laboratory.

2. Maintain cleanliness and orderliness in the laboratory. Refrain from loitering outside the
laboratory room.
3. Disinfect working tables with chlorox, lysol or alcohol before leaving the laboratory. 4.
Treat specimens with disinfectants before disposal. Never flush specimens in the sink. Use
the waste cans in disposing remnants.
5. Make use of the waste cans properly. Strictly separate degradable from non
degradable and infectious from non-infectious wastes.
6. Used slides should be washed or disposed properly. Never leave unwashed slides on
the working tables or in the sink.
7. Return stools under the working tables and pick up pieces of paper before leaving
working area.
❖ The instructions on laboratory reports will be followed regardless of online or face-to-face

instructions
INDIVIDUAL or GROUP REPORTS and QUOTA REQUIREMENTS
1. Whenever a written report is required, references must always be included (Book

title; Author; Edition; Page number) in each item /number of the written report.
Written reports without references will NOT be considered.
2. Online references should contain the complete website address.
3. Record data and label drawings in pen, draw/sketch using pencil (plain or
colored).
4. Submit reports and drawings on time. Late reports and drawings shall be given a
50% deduction from the total score. Reports/drawings submitted on another day
shall be considered for completion purposes only.
5. Margination for written reports are: 1.0 inches – Left; 0.5 inch – top, right & bottom (in
RED ink)
6. Quota specimens should be provided by the students. (Students may obtain
pathologic quota specimens from other clinical laboratories). (as applicable) 7. Slides
and cover slips should also be provided by the students. Reagents will be provided
by the central supply office. (as applicable)
PARASITOLOGY LABORATORY DRAWINGS RUBRIC
❖ The rubrics will be utilized for all laboratory activities integrated in this module EXCEPT for
experiments 3 and 4
CATEGORY 4 3 2 1
Labels Every item that recognizable. All structures are labeled accuracy.
needs to be assigned with 85- 95% accuracy. Less than 75% of the items
identified has a structures are Most items (75- that need to be
label. It is very clear labeled very 89%) that need to be identified have labels. It is
which label goes accurately. identified have labels. It is not clear which label
with which Almost all items (90%) that somewhat clear goes with which item.
structure. need to be identified which label goes Fewer than 85% of the
have labels. It is with which assigned
Drawing All assigned clear which label goes structure. details are
details have been with which structure. Almost all assigned details present.
added. (at least Most details are not clear
The details are very Almost all 85%) have been and are difficult to
clear and easy to assigned added. A few identify.
identify. details (at least 85%) havedetails are not clear and
been added. The are difficult The assigned
details to identify structures are drawn with
are clear and easy less than 50%
to identify. accuracy.
The assigned
Accuracy The assigned The assigned structures The assigned structures
structures are drawn are drawn with 85- 95% are drawn with 50-85% structures are labelled
very accurately. accuracy. All accuracy. with less than 50%
assigned The assigned structures accuracy.
and are
are labelled with 50- 84%
Attention to Detail the drawing and the differences between the more major
There are no specimen structures. between the drawing and the differences
differences between There are 30% or less There are 30-50% specimen between the
obvious obvious differences There are 51% and
information.. drawing.
Work shows attention drawing and the

to detail with specimen structures. Work shows specimen and the
magnification views structures. Work shows errors that are drawing, and overall
minimal attention to represented when details for drawing and
and annotation
detail with comparing the information are missing.
information.
magnification views and specimen with the
annotation
All the format 1-2 of the format 3-4 of the format date, labels and
requirements are requirements are requirements are magnification of the
present. missing. missing. drawing are missing.
Magnification work Magnification work Magnification work Magnification work
Formatting (50%) is done at the back is shown at the back is shown on back of is not shown on
- Title, Name, Date, of the drawing with of drawing with less back of
Labels, more than 75% drawing with 50- 75% than 50% accuracy. drawing.
Magnification accuracy. accuracy. The title, name,
PARASITOLOGY LABORATORY QUESTIONS FOR RESEARCH RUBRIC
CATEGORY 4 3 2 1
Ideas and Content All the information needed to answer the question are present.
Supporting details information followed. question is specific explanation
are included. The needed to answer Less than 50% of present. The in answer to the
directions were the question is the information directions were not question.
followed. present. The needed to followed.
50-80% of the directions were answer the There is no clear or
Conventions Less than or capital letters and No terms from the lesson
10% have end marks or structural and spelling are used.
capital letters and errors. Answer is clear.
structural and Less than 50% of terms
Use of terms 76-100% of spelling errors. from the lesson are used.
the terms from the Answer is very Less than 75% of the
lesson are used. clear. terms are accurately
More than 95% of 50-75% of the used.
the terms are terms from the lesson are Sentences are
accurately used used. 75-95% of the incomplete or too long.
and are fully terms are At least 50% of the
defined. accurately used. sentences are complete
Sentence Fluency More and easy to
than 75% of the understand.
sentences are At least 75% of the More than 50% have end
complete and sentences are complete marks or capital letters
clearly understood. 31-50% have end marks and structural and
and clearly
or capital letters and spelling errors.
Connections with understood.
structural and spelling Answer not clear.
each other are
errors. Answer
clearly established.
somewhat clear.
10-30% have end marks
Documentation incorporated format with D n

and into paper as few citation e d
Referencing appropriate errors m a
Uses Uses Includes o n
appropriate appropriate references from ns o
evidence and references from a few relevant tr v
documentation a number of secondary at er
demonstrating relevant information es re
highly information sources, more p li
competent sources, than one o a
information showing tertiary source. or n
access, adequate Demonstrat d c
evaluation and information es limited o e
application. access, c o
bibliographic
Thoroughly evaluation and u n
research.
evaluates application. m te
Displays
source Notes briefly e rti
inconsistencies
accuracy, issues of source nt ar
about issues of
reliability, and accuracy, at y
source
authority. Cited io so
reliability, and accuracy,
works/bibliograp n ur
authority. reliability, and
hy follow a sk c
Cited works authority. Some
standard style ills es
/bibliography information is
and or .
follow a not referenced.
bibliographic ef Sil
standard style Contains
format and fo e
and several
are skillfully rt nt
bibliographic citation errors.
a a
b so lit n of
o ur y, ci st
ut c a n yl
or e n g e
u a d la a
n c a c n
a c ut ks d
w ur h c /
ar a or o or
e c it ns fo
of y, y. ist r
iss re R e m
u li ef n at
es a er c .
of bi e y
NAME: RATING: Laboratory Schedule: Group No: Date performed: Date submitted:
Experiment No. 1
INSTRUMENTS AND GLASSWARES USED IN CLINICAL PARASITOLOGY
I. Background
The microscope is an essential tool in the study of microorganisms, cells and tissues.
As an instrument, it must be handled and used carefully at all times. In order for the
observer to view organisms under it, it is but necessary that one must know how to use a
microscope effectively.
Aside from the compound microscope, glasswares and instruments are also
needed in Parasitology Laboratory. Glassware is used to contain reagents as well as
parasites for analytical identification. Other instruments are used to facilitate the
diagnosis and identification of parasites.
II. What are expected of you?

At the end of the experiment, you are expected:
1. To describe the different glasswares and different instruments commonly used in
Parasitology
2. To demonstrate the proper operation of the different instruments and glasswares
commonly used in Parasitology
III. What are the materials?
✓ Books
✓ Online resources/references
IV. How do you go about this experiment?

Research on instruments and glass wares. Study the parts, operating techniques
and their uses.
1. Draw and label:

a. Binocular microscope
b. Clinical Centrifuge
c. Glass slides and coverslip
d. Centrifuge tube
e. Plain test tube
f. Beaker
BINOCULAR MICROSCOPE CLINICAL CENTRIFUGE

CENTRIFUGE TUBE PLAIN TEST TUBE
GLASS SLIDE AND COVERSLIP BEAKER
Questions for enrichment (answer on the space provided) 1. What
are the quality control measures for the following: A. Binocular
microscope
B. Clinical Centrifuge
C. Glass slides and coverslip
D. Centrifuge tube
E. Plain test tube
F. Beaker
2. What are the different parts of the binocular microscope? List and indicate the
functions of each.
3. Describe the different glass wares/instruments below and give the function of
each in clinical parasitology.
NAME: RATING:
Laboratory Schedule: Group No: Date performed: Date submitted:
Experiment No. 2
PHYSIOLOGY OF STOOL FORMATION AND PROPER COLLECTION OF SPECIMEN
I. Background
Feces may be defined as the residual mass of material remaining in the
intestinal tract after the full and complete exercise of the digestive and absorptive
functions and are ultimately expelled from the body through the rectum. The
semi-fluid intestinal contents at the duocecal valve are transformed into feces in the
large intestine where the residues remain for one or more days.

1. To enumerate the different organs in the human digestive system

2. To describe the different organs involved in stool formation
3. To demonstrate the proper collection of stool.
Reference materials
Illustration materials/Chart of the Human Digestive System
A. Formation of Feces:
Food is received through the mouth, when food has been chewed to particles
of proper size, mixed with salivary amylase and mucus and shaped into a lubricated
“bolus”; it is now ready for swallowing. The bolus is forced backward through the
mouth and goes down into the pharynx and into the esophagus. Food is forced into
the stomach through the esophagus by muscular movements known as “peristalsis”.
Gentle peristaltic movements pass over the stomach are emptied into the small
intestine. During the passage of intestinal contents through the small intestine,
products of digestion together with many other compounds like vitamins, water,
mineral salts are absorbed. When the contents reach the large intestine, the process
of absorption, with the exemption of water, is normally completed. Here more water
and sodium chloride are absorbed and the remaining material leaves the body as
feces.
B. Proper Collection
1. Stool Container
Stool containers should be covered, clean, wide-mouthed and preferably

colorless. It should be free from water, urine and soil.
2. Time
Submit the specimen immediately to the laboratory and see to it that it is

completely and properly labeled.
Stool examination falls under clinical microscopy in a hospital set-up and thus
may not be given priority since there are other specimens to be examined. However, in
the diagnosis of amoebiasis, consistency of stools dictates that diarrheic or watery
stools must be examined within 30 minutes to 1 hour since trophozoites die within this
period outside the body of the host.
If transportation and examination is to be delayed it should be immediately preserved

in 10%formalin solution or polyvinyl alcohol (PVA).
3. Amount
The amount .of stools to be submitted will depend on the technique that will be
performed. For routine stool examination, 20-40 g of formed stools (usually half of the
thumb) or 5 -6 tbsps of watery stools will suffice.
However, there are situations when it is necessary to submit the whole stool movement
like in cases where the laboratory likes to recover helminthic adults after treatment. This
is best exemplified by the examination done after Taenia species treatment.
V. Draw and label:

1. Human Digestive System
2. Flow chart of stool formation
HUMAN DIGESTIVE SYSTEM FLOW CHART OF STOOL FORMATIONQuestions for enrichment

(write answers on the space provided)
1. What are the importance of proper collection of fecal sample?
2. What is the importance of delivering the stool specimen in the laboratory as soon
as possible?
3. Give the reasons why stool should not be mixed with toilet water and urine.
4. List different IMPROPER collection of fecal sample
NAME: RATING:
Experiment No. 3
ROUTINE STOOL EXAMINATION: PHYSICAL & CHEMICAL PHASE
I. Background
A normal stool consists chiefly of undigested food particles, various products of
digestion and a large amount of bacteria, which are usually non-pathogenic. When
diseases occur, food particles increase since the food material is swept out before
complete digestion or absorption can take place. Many of these food remnants can
be identified either macroscopically, such as seeds, fruit or vegetable skins, etc., or
microscopically, such as undigested muscle fibers, vegetable cells, vegetable fibers
etc.

1. To differentiate the normal and abnormal physical characteristics of stool
2. To be able to perform routine chemical examinations on fecal samples. 3. To

describe and interpret the results of chemical tests performed on fecal samples

✓ Books

Procedure:
A. Physical Examination: take note of the .color, odor and Consistency of the fecal
sample.
Normal: Color - yellow or brown
Odor: offensive but excessively
Consistency: soft to semi-formed
B. Chemical examination:
Occult Blood - Benzidine method
1. Smear a pea size stool on a strip of filter paper

2. Put drops of saturated acetic acid with benzidine powder at the center of your
smear.
3. Add an equal amount of 3% hydrogen peroxide.
4. Read the result not after than 2 minutes after the addition of the last reagent.
Results and Interpretations:
Trace - very faint blue
(+) - Faint blue
(++) - Distinct blue
(+++) - Dark blue
(++++) - prussian blue
V. Results and observations:

Color:___________________________ Odor:_____________________
Consistency:______________________ Benzidine test:______________
VI. Draw and label:

1. Positive and neqative results/ Standard Results of the benzidine
POSITIVE STANDARD RESULT NEGATIVE STANDARD RESULT
Questions for enrichment (write answers on the space provided)
1. In a table form, list down the different colors of stool with their corresponding
clinical significance
STOOL COLOR ASSOCIATED DISEASES
2. In a table form, list down the different stool consistency with their
corresponding clinical significance
STOOL CONSISTENCY
ASSOCIATED DISEASES
3. Explain the importance of the occult blood test. Illustrate through a flow chart
the principle of occult blood.
4. List the causes of false positive and false negative results in occult blood test.
5. What is the patient preparation prior to occult blood test?
6. What is the test to distinguish between the presence of fetal blood or maternal
blood in an infant’s stool or vomitus? Discuss the principle and the expected
results.
NAME: RATING:
Experiment No. 4
ROUTINE STOOL EXAMINATION: MICROSCOPIC
I. Background
The microscopist should be familiar with the different structures found in the feces
such as trophozoites and cysts of Amoeba, helminth eggs and larvae, RBC,
macrophages, WBC, fungi, plant cells (pollen grains and spores), epithelial cells,
crystals like Calcium oxalate and triple phosphate, bacteria, plant fibers, root hairs
and animal cells are similar to helminth ova.
Most common errors in the identification of parasites may be due to the following:
(1) Lack of familiarity with the parasites, (2) Cursory examination of the slide and (3)
Confusion of parasite with artifacts.

To be familiar with the different method and techniques commonly employed in

the identification of parasites using the stool sample

Slides and Cover slips 10% formalin solution
Applicator sticks ether or ethyl acetate
NSS gauze
Lugol's solution Centrifuge tubes
KTS (Cellophane strips soaked in malachite green solution of glycerol)
✓ Books

Procedure:
A. Direct Fecal Smear (DFS) or Direct Wet Mount Technique
This is the simplest and moderately efficient procedure for examinations of feces. The
mixing action of the intestinal tract usually results in an even distribution of parasites in
stool. DFS preparations from soft, loose and watery specimens should be mandatory. It is
highly recommended for the detection of motile trophozoites in liquid, diarrheic and
bloody mucoid specimens, however, if the number of organisms are few, this method
may be of low sensitivity and insufficient to reveal their presence.
1. Comminute approximately 2 mg of feces with a drop of NSS on a clean slide

using an applicator stick until a uniform suspension without fibers or gritty
materials obtained.
2. Place the cover slip above the preparation and immediately examine under
the LPO of the microscope in a systematic method (figure A). Confirm
identification under HPO.
3. Wet mounts may be preserved for several hours by ringing the cover glass with
paraffin or nail polish.
Figure A: method for systematically scanning a wet mount
preparation. Notes:
The amount of feces used for the direct mount is important. The 2 mg recommended
approximates what would form a low cone at the end of a wooded applicator stick.
When less than 2 mg of feces is used, the suspension will be too thin and may have
blank spaces, whereas the use of more than 2 mg results in a suspension that is too thick
and parasites may be hidden under fecal debris.
Temporary stains may be used with wet mount preparations to aid in location and
identification of protozoa but are not necessary for eggs and larvae. Dilute iodine
solutions are the most commonly used temporary stains and are most useful for
recognition of cyst stage; however, they kill and distort the trophozoites. In cysts, the
visibility of nuclei is enhanced so that their number and morphological features are more
clearly seen.
The use of weak iodine solutions is not recommended since they do not stain organisms
well. Likewise, the use of an iodine solution that is too strong will stain the organisms so
darkly that morphological features cannot be seen. Dilute iodine solutions that are
recommended are D' Antoni's, Dobell and O'Connor's and Lugol's.
B. Kato - Thick Smear Preparation:

This technique is based on the fact that when a layer of stool is in contact with glycerin
soaked cellophane for a period of time, it becomes clarified and helminth eggs become
visible. .
1. Place a pea-size stool sample at the center of glass slide and cover with a
square piece of pre-treated cellophane.
2. With the aid of a rubber stopper, press the cellophane gently to spread the stool,
taking care that the specimen does not spread beyond the cellophane cover,
the cellophane also serves as a cover slip.
3. Leave the prepared slide at room temperature for 10-20 minutes. During this time
the microscopic field becomes clear due to the action of glycerin on the stool
constituents.
4. The slide should be examined after 10-20 minutes or within one hour after
preparation. Allowing the slide to stand for long period of time will cause drying
and shells of hookworm ova will become be difficult to see.
C. Formalin - Ether Concentration Technique (FECT)

The use of concentration procedures for fecal examination virtually ensures the
detection of even small numbers of organisms that would go undetected if only direct
smears of permanent-stained slides were examined.
Concentration procedures generally fall into two categories - flotation and
sedimentation. Sedimentation can be achieved by simple gravity or centrifugation. Since
the sediment will generally contain all the parasites occurring in the stool sample, this
procedure has greater diagnostic sensitivity. An added advantage is that the technique
can be readily applied to both fresh and preserved feces.
1. Thoroughly comminute approximately 1.0 to 1.5 g of fresh feces in 10 ml 10% formalin

in a suitable container. Let it stand for 30 minutes or longer to achieve adequate
fixation. For very loose or watery fecal sample, use 5-6 ml of material.
2. Strain the suspension through two layers of wet gauze and pour into a 15 ml conical
centrifuge tube.
3. Fill tube with NSS; centrifuge at 400 - 500 g for 1 – 2 minutes.
4. Discard the supernatant if cloudy; the sediment should be resuspended then
centrifuged again using NSS. Proceed to the next step only after the supernatant is
clear.
5. Resuspend the sediment in 10% formalin to a volume of 10 ml; add 3 ml of ether (or
ethyl acetate) then shake vigorously for 30 seconds. If ether is used, hold tube with
stopper directed away from face.
6. Centrifuge at 400 - 500 g for 2 -3 minutes. .When tube is removed, it will be seen to
consist of four layers:
a. a top layer of ether
b. a plug of debris adhering to the wall
c. a layer of formalin
d. sediment for examination. Insert applicator stick with cottoned tip to ring and
loosen the plug of debris
7. Decant tube, discard top 3 layers, mix small amount of fluid left with the remaining
sediment and pipette out.
8. Prepare iodine and unstained wet mounts for examination
Notes:
Recently, concern over storage and use of ether, a potentially flammable and
explosive material, has led to the use of ethyl acetate as a substitute. Ethyl acetate
appears to be somewhat more efficient than ether when specimens contain Taenia or
Hymenolepis nana eggs or Giardia cysts since there are fewer tendencies for these
eggs or cysts to become trapped in the plug of debris.
However, one disadvantage is that ethyl acetate, when used as a solvent for
fresh feces, is not as efficient as ether in extraction of fatty or mucoid materials which
may interfere with the microscopic examination of the sample
V. Draw and label:

a. Print/ Copy paste and Label:
1. a picture of a DFS stool smear under the microscope
2. a picture of a KTS stool smear under the microscope
b. Draw and Label:
1. Procedures A, B and C
PRINT and LABEL (Microscopic)
DFS
KTS
PROCEDURE A
PROCEDURE B
PROCEDURE C
1. List the reagents used in KTS and the importance of each
2. Discuss the purpose of concentration technique
3. Enumerate and describe the concentration methods
4. Enumerate and describe the different flotation methods
5. Identify the specific parasites that can be isolated in each special method
SECTION 2: MORPHOLOGY, PATHOPHYSIOLOGY, LIFE CYCLE, SPECIMENS USED FOR

IDENTIFICATION, DIAGNOSTIC FEATURES, PREVENTION & CONTROL OF NEMATODES
(ROUNDWORMS)

1. identified each member of Phylum Nematode according to morphology, and
recognize stage of the parasite being studied
2. Valued the importance of ethics of observing patients’ and results’ confidentiality
3. Maintained ethical standard in working with other health personnel
LESSON PROPER
This part begins with the discussion of the helminths. The first group of helminths
discussed are the nematodes, commonly known as the intestinal roundworms.
GENERAL CHARACTERISTICS:
1. General morphology: Unsegmented, elongated and cylindrical in shape. The body is

covered by a non-nucleated cuticle which may be smooth striated, bossed or
ornamented with spines.
• Pseudocoele – body cavity which contains all of the viscera (digestive,
excretory, reproductive and nervous system.
• Anterior end – mouth/buccal cavity which may be provided with spines, hooks,
cutting plates, stylets, or other structures for attachment or penetration • Length –
a few mm to a meter in length
• Movement – by sinuous changes of their bodies
2. The sexes are separate, and the females are larger than the males; male posterior end
is usually curved
3. Reproductive system
• Male: testes, vas deferens, seminal vesicle, ejaculatory duct, cloaca
a) accessory copulatory apparatus
b) gubernaculum (wing- like appendage)
c) telamon
d) copulatory spicule
e) copulatory bursa (umbrella like ex: Hookworms)
4. Digestive System: with complete alimentary tract
• Esophagus: filariform, rhabditiform, spiruroid, strongyliform or stichosoma
5. Nervous System: with chemoreceptors
• Acetyl choline- excitatory neurotransmitter
• Gamma-amino butyric acid (GABA) – inhibitory neurotransmitter
• AMPHIDS: pair of laterally placed minute receptor organs in the cephalic or
cervical region of all nematodes.
• PHASMIDS: pair of minute lateral post-anal organs in species w/o caudal glands 6.
Excretory System: with collecting tubules or collecting canal & an excretory pore 7. No
circulatory system (fluid of the body cavity contains hemoglobin, glucose, proteins, salts
and vitamins)
8. No respiratory system
9. Developmental stages: 1) egg stage 2) four larval stages 3)) adult stage 10.
Reproduction: The adult female may be
a) Oviparous (ex. Ascaris)
b) Viviparous (ex. Trichinella)
c) Parthenogenetic (ex. Strongyloides)
11. Modes of attachment
• Anchorage with attenuated ends (T. trichiura)
• Oral attachment to the mucosa (Ancylostoma)
• Penetration of the tissues (Strongyloides)
• Retention in the folds of the mucosa (lumcricoides)
12. Modes of nourishment
• By sucking with ingestion of blood
• By ingestion of lyzed tissues and blood by embedded worms
• By feeding with intestinal contents

• By deriving from body fluids
13. Life span
o Trichinella spiralis: 4 – 16 weeks
o Enterobius vermicularis: 1 – 2 months
o A. lumbricoides: 12 – 17 months
o Hookworms: at least 14 years
o Filarial worms: up to 25 years
o T. trichiura: 5 – 10 years
o S. stercoralis: 20 – 30 years
14. Life Cycle
o Egg - three layers:
Vitelline membrane
Chorionic or true shell
Albuminous covering
o Four larval stages
o Adult stage
CLASSIFICATION OF MEDICALLY IMPORTANT GROUPS AND SPECIES OF NEMATODES
Class: Aphasmidia (lack “Phasmids” or without caudal chemoreceptors)
A. Species which parasitize the small intestine

1. Trichinella spiralis
2. Capillaria philippinensis
B. Species which parasitize the large intestine
1. Trichuris trichiura
Class: Phasmidia (with “Phasmids”)
A. Species which parasitize the small intestine

1. Ascaris limbricoides
2. Necator americanus
3. Ancylostoma duodenale
4. Ancylostoma ceylanicum
5. Strongyloides stercoralis
B. Specie which parasitize the large intestine
1. Enterobius vermicularis
C. Species which parasitize the tissues
1. Wuchereria bancrofti
2. Brugia malayi
3. Loa loa
4. Dracunculus medinensis
D. Species which causes larva migrans in man

1. Ancylostima braziliense
2. Ancylostoma caninum
3. Toxocara spp.
SECTION 2A: MORPHOLOGY, PATHOPHYSIOLOGY, LIFE CYCLE, SPECIMENS USED FOR

IDENTIFICATION, DIAGNOSTIC FEATURES, PREVENTION & CONTROL OF APHASMID
NEMATODES (ADENOPHOREA)

LESSON PROPER
The Adenophorea (also called the Aphasmidia) seems to be the most primitive group
of nematodes. Mainly, they are free-living in soil and water; however, there are a few
parasitic forms of aphasmids. As the alternate name implies, they do not have phasmids,
and the amphids are located posteriorly on the head region. In fact, they have no sensory
bristles or papillae on the head and body. They are simple, spindle-shaped worms with
simple excretory organs (single-celled).
1. Trichinella spiralis
• Synonym: Trichinella spiralis
• Common name: Trichina worm
• Infective stage: Encysted larva
• Main Habitat: small intestine, skeletal muscles (larva)
• Final Hosts: hogs, rats,
man
• Intermediate hosts: same
animal as the final host
• Developmental stages:
1. Larva: at birth: 80-120 µm
& highly coiled
: has spear-like burrowing tip
at its tapering anterior end
: grows rapidly about 1 mm
2. Adults: rarely seen in stool
or any material
✓ Females: 3.5 mm x 0.06 mm
: Posterior end: bluntly rounded; anterior fifth: with single
vulva
LARVIPAROUS
✓ Males: 1.5 mm x 0.04 mm
: posterior end: ventrally curved with two lobular caudal
appendages

• Life span: 4-16 weeks • Infective
stage: ENCYSTED LARVAE
• Mode of transmission: ingested of
encysted larva
• Life Cycle: BLIND ALLEY CYCLE - requires 2
hosts in order to
complete the life
cycle
• Human infection is a dead-end infection
• Disease: Trichinosis, Trichiniasis, Trichinelliasis

Stages of pathologic changes:
1. Intestinal: inflammation of duodenal & jejunal mucosa, malaise, nausea,

diarrhea and abdominal cramps
2. Muscle invasion: fever, facial edema (particularly in the eyes), pain, swelling
& weakness of the involved muscle, eosinophilia
3. Convalescence: begins at about the end of the 3rd week; s/s subside; cyst
wall then the larva itself CALCIFY
3 clinical phases
1. Intestinal Phase
✓ first week
✓ small intestinal edema and inflammation
✓ nausea, vomiting, abdominal pain, diarrhea, headache and fever,
myalgia
2. Migration Phase
✓ up to 6th week:
- high fever (40oC)
- blurred vision
- edema of the face and eyes
- cough, pleural pains
- eosinophilia (15-40% for 1 month)
✓ 4th to 8th week: death
3. Muscular Phase
✓ acute local inflammation
✓ edema and pain of the musculature
✓ Larval encystation
✓ muscle fibers
- 3-4 days after larval invasion
- Edematous
- spindle shape
- lose their cross striations

- undergo basophilic degeneration
- Nuclei:
size and number
migrate to the anterior of the muscle cell
Predominating symptoms
✓ Eosinophilia
✓ orbital edema
✓ muscular pain and tenderness
✓ shallow, painful breathing
✓ general weakness
4. Encystment/Encapsulation Phase
✓ fever, weakness, pain and other symptoms start to abate
• Diagnosis
1. Muscle Biopsy:
✓ gastrocnemius and biceps
✓ digest with pepsin-hydrochloride
✓ 3rd or 4th week of infection
2. Bachman Intradermal Test
✓ use antigen preparation of Trichinella larva
✓ Observe after 30 minutes
✓ (+) result: large elevated swelling at the site of injection
3. Beck’s Xenodiagnosis
4. Serodiagnosis
5. Bentonite Flocculation
6. Blood Count : Eosinophilia
• Prevention:
✓ Elimination of encysted larva in hogs through freezing at -30oC for 24 hours
✓ Extermination of rats and mice around farms
✓ Thorough cooking of pork
2. Capillaria philippinensis
• Common name: Pudoc worm

• Infective stage: 3rd stage larva
• Intermediate host: glassfish, “bagsit”, “bagsang”, “ipon”
• DH: Man and birds
• Main Habitat: L.I. & S.I.
• Distribution: Philippines & Thailand
• Developmental stages
1. OVA
✓ Color: Pale yellow

✓ Size: 42x20 µ
✓ similar to that of T. trichiura
✓ smaller & more striated shells
✓ Flattened plugs
✓ Peanut shape
2. FEMALE
✓ size: 2.4 – 4.3 mm
✓ Divisions:
✓ Anterior – esophagus and esophageal glands
✓ Posterior – intestine and reproductive organs
❖ Atypical female
- uterus lined with 2-3 rows of eggs
- Larviparous
- causes internal auto-reinfection
❖ Typical female
- uterus lined with 1 row of egg
- oviparous
3. MALE
✓ size: 2.3 – 3.17mm
✓ caudal alae; long, non-spiny sheath
• Disease caused: Capillariasis or Mystery disease
• Predominant Symptoms
✓ borborygmi
✓ abdominal pain
✓ Diarrhea (chronic)
✓ Untreated:
✓ weight loss; malaise
✓ vomiting; dehydration
✓ anorexia
✓ pneumonia, heart failure and cerebral edema
✓ Death: 2 – 8 weeks after these are seen
• Treatment
1. Albendazole
✓ drug of choice
✓ 400 mg/day for 10 days
✓ destroys larvae readily
2. Mebendazole
✓ 200mg twice a day for 20 days or 400 mg/day for 20 days
3. Electrolyte replacement therapy and high protein diet
• Prevention: Thorough cooking of fish

• Laboratory Diagnosis: DFS & Concentration technique
• Mode of Transmission: eating infected fish
3. Capillaria hepatica
• Common name: Capillary liver worm

• Infective stage: embryonated ova
• Hosts: rats, dogs, cats, monkeys and rarely in human
• Habitat: liver of the host
1. Ova:
✓ “lemon – shaped” eggs
✓ resembles that of Trichuris trichiura
✓ outer shell: pitted like golf ball
2. Adults
✓ males are half as long as the female with slightly chitinized spicule
• Diagnosis: Liver biopsy (revealing characteristic egg and parasite) •
Transmission:
✓ Eating of dog’s meat
✓ Ingestion of contaminated food and drinks (embryonated ova)
4. Trichuris trichiura
• Synonyms: Trichocephalus trichiurus, Trichocephalus dispar
• Common name: whipworm
• Principal host: man, but has been found in hogs, monkeys, cattle, dogs,
mice
• Main habitat: cecum & appendix
• Life span: 5-10 years
1. Ova:
✓ Barrel/football-shaped, Japanese lantern
✓ 50 µmX25µm in size
✓ 3 layers:
✓ undeveloped, unicellular embryo
✓ outermost layer – smooth, bile-stained
✓ Hyaline/Mucus plug
2. Adult
✓ flesh – colored
✓ anterior three-fifths is attenuated (whiplike)
✓ stichosoma type of esophagus
a) male
- 30-45 mm
- posterior portion: coiled ( 360o)

- lanceolate spicule protruding through a refractile
pineal sheath
b) Female:
- 35-50 mm
- bluntly rounded posterior end
- 3,000-10,000 eggs per day
• Disease:
✓ Trichuriasis
✓ Trichucephaliasis
✓ Whipworm infection
- Slight infection – asymptomatic
- Heavy infection – surface of colon matted with worms
• Pathology & Symptomatology
✓ Asymptomatic for light infections
✓ Heavy infection: surface colon is matted with worms
- Bloody or mucoid diarrhea
- Weight loss and weakness
- Abdominal pain and tenderness
- Increased peristalsis and rectal prolapse
- Obstruction and inflammation of the appendix (appendicitis)
- Hypoalbuminemia and IDA
- Extreme cachexia
• Diagnosis: DFS, KTS, Concentration techniques
• Treatment: Mebendazole (500mg) , Albendazole (400 mg) & Oxantel-pyrantel •
Prevention
- Sanitary disposal of feces
- Thorough washing of hands
- Thorough washing and cooking of food
- Avoid using human feces as fertilizer
5. Dioctophyma renale
• Synonym: Eustrongylus gigas

• Common name; Giant Kidney Worm
• Infective stage: 3rd stage larva
• Habitat: Kidney (typically the right kidney)
• Hosts:
✓ Definitive - Mink, dogs, foxes, and other carnivores; fish eating mammals ✓
Intermediate - Oligochaete annelids
✓ Paratenic - Fish and frogs.
• Life span: 5 years
• Diagnosis: eggs in urine (Urine sedimentation)
• Treatment: surgical excision
1. Ova:
✓ ellipsoidal and brownish-yellow
✓ deeply sculptured depressions
2. Adults:
✓ blood red in color
✓ attenuated (slightly) on both ends
a) Males:
- Size: 14-20 cm x 4-6 mm
- Bell-shaped copulatory bursa
- not supported by rays; covering of papillae
b) Females
- vulva: midventral near anterior
SYNTHESIS
Aphasmids are nematodes which lack phasmids or it lacks caudal chemoreceptors.

The nematodes in this classification are:
✓ Trichinella spiralis
✓ Capillaria philippinensis
✓ Capillaria hepatica
✓ Trichuris trichiura
✓ Dioctophyma renale
ASSESSMENT
QUIZ
✓ For offline and weak connectivity, it will be provided by the instructor in essay form ✓
For Online/Strong connectivity, it will be provided either through a worksheet, or quizziz or
google forms.
REFERENCES:
Belizario, V. Jr. (1998). Philippine Textbook of Medical Parasitology (1st edition). The
Publications Program.
Zeibig, A. (2013). Clinical Parasitology: A Practical Approach. (2nd ed.). Saunders Elsevier Inc.
Aguinaldo, A. M. A., J. M. Turbeville, L. S. Linford, M. C. Rivera, J. R. Garey, R. A. Raff, and J. A.

Lake. 1997. Evidence for a clade of nematodes, arthropods and other moulting
animals. Nature. 387:489-493.
Diesing (n.d). Description of the Phylum Nematoda. Retrieved from

https://comenius.susqu.edu/biol/202/animals/protostomes/ecdysozoa/nematoda/ne
m atoda-description.html on July 10, 2020
LABORATORY
NAME: RATING:
Laboratory Schedule: Group No:

Date performed: Date submitted: Experiment No. 5
PHYLUM NEMATODA - CLASS ENOPLEA (ADENOPHOREA; APHASMIDEA)
I. Background
Papillae, amphids and phasmids are the main sensory organs of nematodes. Most sensory organs
are found in the anterior end, but males often have well-developed sensory structures associated with
copulation. Phasmids are paired structures found posteriorly. The presence or absence of phasmids had
been used to classify nematodes into two classes.
In class Adenophorea; Aphasmidea, the amphids are generally well developed (except in parasitic
forms), well behind the lips that are often pocket like. Caudal and hypodermal glands are common.
Phasmids are absent. The excretory system lacks lateral canal and are formed of single, ventral,
glandular cells, or entirely absent. Dierids are absent. Most of the members are free living and some
are parasitic on plants and animals.
General characteristics of the parasitic species within the Enoplea:
A. with 5 esophageal glands

B. generally with caudal and hypodermal glands
C. phasmids absent
D. excretory system formed of single, ventral, glandular cells or may be entirely absent
1. To know the morphology, life cycle, and laboratory methods employed in the identification of
Nematodes.
2. To be familiar with the diagnostic and infective stages of these nematodes.
3. Identify the nematodes using the distinguishing marks of their diagnostic stages
4. to know the mode of transmission of these nematodes
Colored plates
Reference Book
a. Print and label: Microscopic (HPO)
1. ovum of Trichuris trichiura
2. ovum of Capillaria philippinensis
3. encysted larvae of Trichinella spiralis
b. Draw and label: Diagrammatic
1. Adult male and female (typical and atypical) Capillaria philippinensis
c. Label the parts: Diagrammatic
1. Adult male and female Trichuris trichiura
2. Adult male and female Trichinella spiralis
PRINT and LABEL ( Microscopic)

ovum of Trichuris trichiura ovum of Capillaria philippinensis
encysted larvae of Trichinella spiralis
DRAW and LABEL (diagrammatic)
Adult male (typical and atypical) Capillaria philippinensis
Adult female (typical and atypical) Capillaria philippinensis

LABEL THE FOLLOWING:
1. ADULT MALE and FEMALE Trichuris trichiura

lectures.blogspot.com/2016/07/lecture-2-nematodes.html)
A. ADULT FEMALE
1. _____________________ 2.
_____________________ 3.
_____________________ 4.
( http://mt _____________________ 5.
_____________________ 6.
_____________________ 7.
_____________________ 8.
_____________________ 9.
_____________________
B. ADULT MALE
1. _____________________ 2.
_____________________ 3.
_____________________ 4.
_____________________ 5.
_____________________ 6.
_____________________ 7.
_____________________ 8.
_____________________
2. ADULT MALE and FEMALE Trichinella spiralis
(https://www.pinterest.ph/pin/558164947550122566/)
MALE
1. ______________________________
2. ______________________________
FEMALE
1. _____________________________
2. _____________________________
3. _____________________________
4. _____________________________
5. _____________________________
6. _____________________________
7. _____________________________
8. _____________________________
9. _____________________________
1. In a table form, differentiate Trichuris trichiura, Capillaria philippinensis and Trichinella spiralis based
on: ova, morphology, diagnostic stage, infective stage, other name, disease caused, distinguishing
mark or characteristic, mode of transmission
2. Describe the life cycle of Trichuris trichiura, Capillaria philippinensis and Trichinella spiralis
3. Give and describe the laboratory methods in the identification of Nematodes.

SECTION 2B: MORPHOLOGY, PATHOPHYSIOLOGY, LIFE CYCLE, SPECIMENS USED FOR

IDENTIFICATION, DIAGNOSTIC FEATURES, PREVENTION & CONTROL OF PHASMID
NEMATODES (PHASMIDEA)

LESSON PROPER
The Secernentea (also called the (phasmidia) vary greatly in size from microscopic to
several feet long. The largest known secernentean, which is up to 30 ft (9 m) in length, lives in
the placentas of female sperm whales. The body of secernenteans consists of a flexible
cylinder that tapers at both ends, with a pointed tail and a blunt head. They are considered
non-segmented pseudocoelomates; that is, creatures possessing a three-layered body that
has a fluid-filled body cavity (pseudocoelom) between the endoderm and the mesoderm
(the innermost and middle tissue layers).
A flexible but tough collagenous cuticle surrounds the body with a system of grooves
across the body from head to tail, which protects them internally. The non-cellular cuticle
varies from four to two layers and is almost always transversely striated. Laterally for most of
the body length, the cuticle is generally modified into a wing area that is marked by
longitudinal ridges; generally, this region is only slightly above the normal body contour.
However, in some parasitic forms, it may extend out a distance equal to the body's diameter.
The cellular hypodermis is the subcuticular layer that secretes the cuticle.
The sensory system contains phasmids, which are a pair of bilateral cuticular, glandular
organs, situated laterally in the caudal (posterior to the anus) region and opening to the
surface by a slit or pore. Also known as precaudal glands, phasmids are unique to the
secernenteans, in which their function is believed to be sensory. At the other end are pore
like amphid apertures, which are a pair of glandular chemosensory organs situated dorso
laterally in the cephalic (head or anterior) region and opening through the cuticle. Although
usually pore-like, in isolated instances the aperture can be an oval or a cleft. The apertures
show little variation throughout the secernenteans. The amphids are always labial (located
on the lips). The external amphidial aperture is usually less well developed than in
adenophorean worms.
Somatic and cephalic setae, which are elongated structures jointed with the cuticle,
are rare. When present, the cephalic sensilla are located on the labial region, and they are
pore-like or papilliform. In males, there may be caudal setae. In females, somatic setae are
absent. Generally, sixteen sensilla are present in the shape of two circles (an inner circle of
six, and an outer circle of 10). In some parasitic groups, the number of cephalic sensilla may
be reduced. Deirids, pairs of pore-like sensilla that usually protrude above the surface of the
cuticle, are usually present on the cervical region near the level of the nerve ring.
Secernenteans contain no hypodermal glands, but the hypodermal cells of the

hypodermis (a thin tissue layer beneath the cuticle that thickens to form the dorsal, ventral,
and two lateral hypodermal chords, and extends the length of the body) are usually
multinucleate (syncytial: more than two nuclei), but may also be uninucleate (cellular: one
nuclei). These divide the muscle cells into four fields. A layer of longitudinal muscles underlies
the hypodermis.
Bursae, or caudal alae, are sublateral projections generated by a longitudinal

widening of the cuticle. It is common for them to be present within male secernenteans,
each looking like a fluid-filled body sac. The thin cuticle extensions are located on both sides
of the anus, specifically on either side of (or sometimes surrounding) the cloaca (the
urogenital opening) of males. The well-developed excretory system is in the shape of an H or
U. It is a simple tubular system that is located in one or both of the lateral hypodermal chords,
and embedded between the three cell bodies in the hypodermal chord. The system opens
ventromedially by way of a cuticularized duct. The rectal part of the gland system is usually
present. They have no caudal glands.
The muscular esophagus or pharynx varies in configuration, but the majority of

secernenteans have three esophageal glands: two that are subventral and one that is
dorsal. The subventral glands open into the posterior metacarpus. The dorsal gland opens
anteriorly into the procorpus or the anterior metacarpus. Its basic structure is corpus (the
anterior part is cylindrical), with the basal (bottom) region sometimes swollen in the shape of
a bulb. The glands empty their contents into the esophagus to aid in digestion. The tail is the
region between the anus and the back tip of the body.
1. Ascaris lumbricoides
• Most common intestinal roundworm

• Common name: Giant Intestinal roundworm
• Definitive host: man (no intermediate host needed)
• Main habitat: Lumen of the SI
• Life span: 12-17 months
• Developmental stages:
1. Ova/Eggs
Types of eggs
a) Unfertilized: longer and narrower
2 layers of the egg shell
✓ albuminoid layer (absent in old specimens)
✓ chorionic layer or true shell
➢ filled with amorphous mass
➢ lack the cresentric clear area
b) fertilized: broadly avoidal and thick
3 layers:
✓ Chorionic/true shell: chitinous layer; secretory product of the
egg
✓ Vitelline layer: fertilization membrane; highly impermeable
membrane that protects the inner embryo
✓ Protein coat/Albuminous layer: outermost mamillated layer with
a tanning action
➢ Embryonated: same as fertilized but contains the larva of the
embryo
➢ Decorticated: lacks the albuminous mamillated shell; usually seen
in old specimens; it may be fertilized or unfertilized
2. Adult: white, creamy or pinkish yellow when freshly expelled and resembles
earthworm (lumbricus); head is provided with three conspicuous lips
which are finely denticulated; each lip has minute twinned sensory
papillae.
a) Male
➢ 10-31 cm
➢ Usually shorter and slender
➢ ventrically curved posterior end with 2 spicules
➢ genitalia: composed of a single, long tortuous tubule
b) Female
➢ 35 cm long x 3-6 mm
➢ straight posterior end
➢ paired reproductive organs located in the 2/3 of the body
➢ oviparous
➢ gravid uterus : 200,000 eggs
• Disease: Ascariasis, Dooryard or Backyard Infection
• Pathology
a) Due to larval migration:
✓ Ascaris pneumonitis: Damage to the pulmonary tissue
(petechial hemorrhage) when larvae break out of the lung
capillaries into the air sacs
✓ Symptoms manifested: asthmatic type of respiration; cough;
bronchial rales (abnormal respiratory sound); urticarial rash
(hives, vascular reaction of the upper dermis; eosinophilia in the
circulatory blood)
b) Due to adult worms:
✓ Diarrhea, vague abdominal pain, nausea and loss of appetite
✓ Due to its erratic behavior: vomiting; suffocation; intestinal
obstruction, appendicitis; acute pancreatitis; peritonitis
(perforation of the bowel)
• Diagnosis: DFS, KTS, Concentration Technique, ELISA
• Stool examination may give negative results due to the following
- During the early stage of infection (worms are still immature)
- During larval migration through the blood stream
- When only male worms are present in the intestines
• Prevention:
✓ Sanitary disposal of human excreta
✓ Personal hygiene
✓ Avoid the use of night soil fertilizer
✓ Thorough cooking of food particularly vegetables and washing of fruits
✓ Washing solution: aqueous iodine solution (200 parts/million)
✓ kills infective egg and larva in 15 minutes
• Treatment: Mebendazole (500mg), Pyrantel pamoate (10 mg/kg (maximum of
1 g)), Albendazole (400 mg)

Clinical Parasitology Module: For The Use of University of Baguio School of Natural Sciences Students Only

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Clinical

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Parasitology is an essential component of clinical laboratory medicine.

In spite of the many advances in technology that has been developed in

COURSE TITLE AND DESCRIPTION

ABOUT THE MODULE

A Self-regulated Learning Module 4

Control of Protozoa. It also deals with the characteristics and role of

Student’s attendance in the online learning sessions is a basic requirement

Requirements of the Course

A Self-regulated Learning Module 6

c. Stay motivated. Your future depends on what you do today. Maintain a

e. Communicate properly. Promptly respond to notifications by regularly visiting

▪ Respect private hours. I do not always open my laptop/email/messenger

▪ Be patient. Messages received between 8 AM to 8 PM will be responded

g. Live lecture/Video conferencing guidelines:

- Wear appropriate clothing and set your gadget in an appropriate

A Self-regulated Learning Module 7

- Log in using your UB gmail account. Unidentified names like nicknames,

- Mute your microphone as soon as you log in to the platform to avoid

- Respect privacy. Do not take a screenshot, picture, snapshott, etc. of

Teresa N. Villanueva, RMT, MACT

2 Continue Section 1 Introduction to Online Lecture, Assessment & Quiz

3 Section 2: Nematose Intro Online Lecture: Assessment &

A Self-regulated Learning Module 9

A Self-regulated Learning Module 10

LEARNING ENGAGEMENT 1 CLINICAL PARASITOLOGY: OVERVIEW

SECTION 1: INTRODUCTION TO PARASITOLOGY

COURSE LEARNING OUTCOMES:

At the end of this session, the students must have:

1. used the basic terminologies and theories related to Parasitology 2.

1. Parasitology 4. Commensalism 5. Mutualism

Incidental parasite 18. Intermediate host

11. Temporary parasite 19. Accidental host

12. Permanent parasite 20. Paratenic host

13. Pathogenic parasite 14. 21. Dead-end host

Pseudoparasite 22. Reservoir host

A Self-regulated Learning Module 12

There are several types of parasites that may be members of a parasite-host

Exposure is the process of inoculating and infective agent, while

A Self-regulated Learning Module 14

A Self-regulated Learning Module 15

A Self-regulated Learning Module 16

F. DISEASE PROCESSES and SYMPTOMS

A Self-regulated Learning Module 17

G. PREVENTION and CONTROL

III. LABORATORY METHODS FOR DIAGNOSIS

A. SPECIMEN FOR DIAGNOSIS

5. Cerebrospinal fluid- Acanthamoeba species

- Too red: alkaline pH

A Self-regulated Learning Module 20

Thick Blood Film:

✓ Infected erythrocytes are counted in relation to a predetermined number of

# of parasites/µl = # of parasites x 8000

• If parasites are <10, 500 WBCs should be counted:

# of parasites/µl = # of parasites counted x 16

A Self-regulated Learning Module 21

✓ Earle and Perez method

Thin Blood Film:

+ = 1–10 per 100 thick fields

++ = 11-100 per 100 thick fields

+++ = 1–10 per thick field

++++ = >10 per thick field