b.

Eyepiece Tube - eyepiece holder, carries the eyepiece,

flexible in binoculars, and can be rotated for max
INTRODUCTION visualization for variance in distance
Parasitology - deals with the study of human parasites which c. Diopter Adjustment - control knob present only in
are of medical importance. binocular that’s used to change focus on one eyepiece
Microscope - used to visualize very minute objects, such as d. Nosepiece/Revolving Turrets - movable circular
cells and microorganisms, giving a contrasting image that is structure that houses objective lens, connected to the
magnified. It is made up of lenses for magnification, each with body tube and lies above the stage
its own magnification powers; depending on the type of the e. Stage - section where specimen is placed, have stage
lens, it’ll magnify the specimen according to its focal length clips that holds the specimen slide in place
f. Stage Control Knob - used to move the stage
3 Structural Parts of Microscope: mechanically. There are two knobs; one for moving left
a. Head/Body Tube?Eyepiece Tube - cylindrical metallic and right and the other for moving forward and
tube that holds the eyepiece lens at one end and backward. This will move the slide in the field of vision
connects the nosepiece at the other end. g. Aperture - hole in the microscope where light pass
b. Arm - connects the base to the head and the eyepiece through
tube to the base, supports the head of the microscope h. Microscopic Illuminator - light source; in some
and is also used when carrying the microscope compound microscope, a mirror, w/c reflects the light
c. Base - lowermost part that supports the entire from an external source is used
microscope structure i. Condenser - collect and focus light from illuminator,
found under the stage
Parts of the Microscope: j. Diaphragm/Iris - control the amount of light that
a. Eyepiece/Ocular Lens - closest to the viewer’s eye, reaches the specimen; adjustable apparatus, hence
located at the top, used to look at the specimen controlling the light intensity and the size of beam light
k. Condenser Focus Knob - moves the condenser
up/down, thus controlling the focus of light on specimen
 l. Rack stop - controls how far the stage should go ● Patients must submit approx. 3-5g of stool collected in
m. Light Switch - electrical control device that’s used to a sterile container
switched the microscope on/off ● Patients should be instructed about preventing
n. Brightness Adjustment - controls the voltage supplied contamination of specimen w/ urine, and that sample
to light bulb, controlling the intensity of light shouldn’t be taken from the toilet bowl
● Specimen should be submitted immediately to the
Types of Objective Lenses laboratory
a. Scanning - red stripes, 4x magnification ● The lab. personnel in turn should promptly process the
b. LPO - yellow stripes, 10x - 20x, effective at generating sample upon receipt
good view of specimen ● In the event of delayed processing, preservatives can be
c. HPO - blue stripes, 40x, used to look at smaller bacteria added to stool samples to maintain its integrity and
and cell structure viability of the organisms for examination
d. OIO - white stripes, 100x, rarely used because you’re Specimen Rejection Criteria
reaching the physical limits of magnification ● Specimen contaminated with urine, water, soil, or other
extraneous material
Adjustment Knobs
a. Coarse Adjustment Knobs - focusing the image under Specimen:
low power magnification; rapidly ● Collect stool in a clean, dry leak-proof container within
b. Fine Adjustment Knobs - fine adjustments, smaller 30 minutes to 1 hour after voiding. If it takes longer,
knob, used to move the stage up/down very slowly specimens must be transported in ice.

SPECIMEN COLLECTION AND PROCESSING OF PRESERVATIVES FOR STOOL SPECIMENS

HUMAN BIOLOGICAL SAMPLES
1. 10% Formalin
A. STOOL
ADVANTAGE
 ● All-purpose fixative ● Not suitable for some permanent smears stained w/
● Easy to prepare trichrome
● Long shelf life ● Inadequate preservation of morphology of protozoan
● Good preservation of morphology of helminth eggs, trophozoites
larvae, protozoan cysts, and coccidia ● Iodine interferes w/ other stains and fluorescence
● Suitable for conc. procedures and UV fluorescence ● Iodine may cause distortion of protozoa
microscopy
● Suitable for acid-fast, safranin, and chromotrope stains 3. Low Viscosity Polyvinyl-Alcohol (LV-PVA)
● Compatible with immunoassay kits and UV
fluorescence microscopy ADVANTAGE
● Good preservation of morphology of protozoan cysts
DISADVANTAGE and trophozoites
● Not suitable for some permanent smears stained w/ ● Easy prep. of permanent smears stained w/ such as
trichrome trichrome (solution both preserves organisms and makes
● Inadequate preservation of morphology of protozoan them adhere to slides)
trophozoites ● Preserved samples remain stable for several months
● Can interfere with PCR, especially after extended
fixation time DISADVANTAGE
● Inadequate preservation of morphology of helminth
2. Merthiolate-Iodine Formaldehyde (MIF)⁸ eggs, larvae, coccidia, and microsporidia
● Contains mercuric chloride
ADVANTAGE ● Difficult and expensive to dispose
● Components both fix and stain organism ● Difficult to prepare in the lab
● Easy to prepare ● Not suitable for conc. procedures
● Long shelf life ● Can’t be used with immunoassays
● Useful for field surveys ● Not suitable for acid-fast, safranin, and chromotrope
● Suitable for conc. procedures stains

4. Sodium Acetate-Acetic-Acid-Formalin (SAF)

DISADVANTAGE
ADVANTAGE ADVANTAGE
● Suitable for both conc. procedures and prep. of ● Permanent smears can be made and stained with
permanent stained smears trichrome
● Easy to prepare ● Zinc is preferred over copper
● Long shelf life ● No mercuric chloride
● Suitable for acid-fast, safranin, and chromotrope stains
● Compatible with immunoassay kits DISADVANTAGE
● Staining not consistent
DISADVANTAGE ● Organism morphology may be poor
● Requires additive (albumin-glycine) for adhesion of ● Copper - morphology of cysts and trophozoites is poor
specimen to slides ● Zinc - better morphology but not comparable to
● Permanent stains not as good as w/ PVA or Schaudinn’s LV-PVA
fixative
7. One-vial fixatives (Ecofix, Parasafe, Unifix,
5. Schaudinn’s Fixative Proto-fix, SFX, and others that may be available)

ADVANTAGE ADVANTAGE
● Good preservation of morphology of protozoan cysts ● Concentrate and permanent smear can be made out of
and trophozoites one vial
● Easy prep. of permanent stained smears ● Immunoassays can be done on most
● No mercuric chloride
DISADVANTAGE
● Less suitable for conc. procedures DISADVANTAGE
● Contains mercuric chloride ● Certain one-vial fixatives must use certain stains
● Inadequate preservation of morphology of helminth ● Color difference of stain
eggs and larvaes, coccidia, and microsporidia ● Staining not always consistent
● Poor adhesion of liquid or mucoid specimen to slides ● Sometimes more expensive than formalin and LV-PVA

6. Modified PVA copper/zinc PROCESSING OF STOOL SAMPLES

A. DIRECT FECAL SMEAR
 B. CELLOPHANE - COVERED THICK SMEAR - Pea-sized fresh stools
(KATO-KATZ TECHNIQUE)
C. CONCENTRATION TECHNIQUES Procedure:
1. Put 1 drop of NSS on the glass slide. Carefully emulsify
DIRECT FECAL SMEAR the sample in a rotary motion using applicator stick
❖ Simple, rapid procedure for diagnosing common 2. Apply a coverslip. Prevent bubble formation by moving
intestinal parasitic infection the coverslip towards the emulsion until they meet, then
❖ Commonly called as fecalysis/stool examination lower the coverslip gently on the emulsion
❖ Routine procedure of stool examination useful in 3. Remove excess fluid on the side of the coverslip by
detection of motile protozoan trophozoites touching the edge of the film with the corner of a piece
of tissue paper to absorb excess fluid
Advantage: ● Lugol’s iodine may be added at the edge of the coverslip
- Easier to prepare to allow the iodine to mix well with the fecal smear. The
- Small amount of stool is needed use of this solution will facilitate better and clearer
- Mixed w/ isotonic soln and examined microscopically viewing of the protozoan cysts
4. View the fecal smear under the microscope using LPO
Disadvantage: magnification. Examine systematically for the presence
- Light infection may be misdiagnosed because only of eggs, larvae, and protozoan cysts
small amount of sample is obtained ● Systemic examination starts at the lower edge of the
smear, then move one field to the right and continuously
Materials: examine each field reaching the upper edge. Make sure
- Clean glass slides that the entire slide has been viewed and examined
- Applicator sticks
- Coverslip KATO-KATZ TECHNIQUE
- Dropper
- Lugol’s iodine soln ★ Kato Technique (Kato Katz and Kato Thick) is used to
- Normal saline solution enhance the morphologic details and inc. the chances of
- Compound microscope isolating ova of the different parasites
★ Glycerin is used to clear all the fecal debris so that the
Specimen: ova become more visible during the microscopic
 analysis
★ Kato katz is the quantitative method that requires a Procedure of the Preparation and Examination of Stool
template for a more accurate reporting while Kato Specimen by Kato-Katz Technique:
Thick is the qualitative method 1. Place a small mound (thumb size) of fecal material on a
★ A quantitative method which allows the detection of all paper. Put the wire mesh/screen on top of fecal
helminth eggs occurring in feces (e.g. Ascaris 2. Using an applicator stick, press the screen over the fecal
lumbricoides, Schistosoma japonicum, Trichuris so that some of it will be sieved through and accumulate
trichiura, Strongyloides stercoralis, Enterobius on top of the screen. Scrape a flat sided spatula across
vermicularis, etc.) the upper surface of the screen so that feces accumulate
★ Developed in 1954 by Dr. Kan Kato and Dr. on the spatula
Momoshige Miura 3. Place a template w/ a hole on the center of the
★ Developed to detect Helminth egg in feces, including pre-labeled glass slide. Fill the hole w/ feces from the
those soil-transmitted and intestinal Schistosoma spatula
4. Using the side of the spatula, pass over the template to
Materials: remove excess feces from edge of the hole
- Glass slides 5. Remove the template carefully from the slides so that a
- Pre-soaked (at least 24 hours) green cellophane in cylindrical of feces is left completely. Dispose the
glycerol (50% distilled H2O + 50% glycerol) - add template and applicator stick in an appropriate
malachite green solution if using transparent infectious waste bin
cellophane 6. Cover the fecal material with a cellophane strip
- Nylon or stainless steel mesh wire pre-soaked in glycerol solution (w/ or w/out malachite
- Spatula/ Tongue Depressor green)
- Applicator stick 7. Invert the preparation (w/ the fecal matter facing down)
- Template (50 mg) 9 mm x 1 mm or (41.7 mg) 6mm x on a paper placed on a smooth hard surface (or another
1.5 mm glass slides), and press it to spread the feces evenly to
- Forceps the edges of the cellophane
- Flat bottom jars for soaking stain 8. Remove the slide carefully by gently sliding it sideways
- Pencil or maker to avoid separating the cellophane strip or lifting it off.
- Record logbook Place the slide on the bench with cellophane upwards.
- Compound microscope Water evaporates while glycerol clears the feces. A well
 made preparation allows newsprint to be read through collected in 10 days. One exception is in the diagnosis
the smear of amebiasis w/c up to 6 specimen in 14 days is
● For all, except hookworm eggs, keep slide for one or acceptable
more hours at ambient temperature to clear the fecal ● Medication and substances may interfere w/ detection of
matter prior to examination under the microscope. For parasites. Stool samples from patients whose therapy
faster clearing, the slide can be placed in 40 degree includes barium, bismuth, or mineral oil should be
Celsius incubator for 30 minutes, or under direct collected prior to therapy or not until 5-7 days after
sunlight for several minutes completion of therapy. If the samples are taken during
● Examine the smear in a systematic manner the course of therapy, these interfering substances may
★ Ascaris lumbricoides and Trichuris trichiura eggs will mask possible parasites
remain visible and recognizable for many months in ● Collection of specimens from patients who have taken
these preparations antibiotics/antimalarial medications should be delayed
for 2 weeks
● Collected in a clean, watertight container with a
tight-fitting lid. The amount should be 2-5 g/size of
walnut
● Toilet paper may mask parasite or make examination
difficult
● Should be labeled with patient’s name and identification
number, physician’s name, and date and time of sample
● To demonstrate the motility of protozoan trophozoites, a
fresh specimen is required. Trophozoite stage is
sensitive to environmental changes and, on release from
STOOL SPECIMEN the body, disintegrates rapidly
● Parasites are often shed intermittently, they may not ● Other parasite stages (protozoan cysts, helminth eggs
appear in a stool specimen on a daily basis; therefore, and larvae) are not sensitive and can survive for longer
multiple specimen are recommended for adequate periods outside the host. Trophozoites are usually found
detection in liquid stool, recommended that liquid specimens be
● Typical stool specimen consists of three specimens, one examined within 30 minutes of passage
specimen collected every other day or a total of three ● In keeping stool with consistency, semi-formed
 specimens may yield a mixture of protozoan cysts and ● Never keep stool samples in incubator. 37 deg C beyond
trophozoites and should be evaluated within of passage 30 minutes destroys amoeba
● Formed stool specimens are not likely to contain
trophozoites; therefore, can be held for 24 hours.If
guidelines weren’t met, specimen should be placed into
a preservative
● 3:1 ratio

STOOL PRESERVATION
● If stool is processed within 1 hour, it may be stored at
room temperature
● Formed stool may be refrigerated 1-2 days if
examination is delayed but it’ll not guarantee recovery
of parasites
● Never freeze the sample
● Trophozoites from a refrigerated stool can regain
motility in warm saline on a warm slide
CONCENTRATION TECHNIQUES
● Increase the chances of detecting and diagnosing
parasites in case of light infection
● Advantage: possibility of separating the parasites from
interfering materials (e.g. bacteria, fibers, or undigested
food particles) that may be present in the sample
● Water/liquid or diarrheic stool containing trophozoites is
not recommended

I. SEDIMENTATION
● Concentrating parasite stages usually present in a large
amount of stool samples about 2g of sediment.
● Have a higher specific gravity in terms of mass and
volume compared to soln and thus will sink to the
bottom

A. Methods
● Acid Ether Concentration technique (AECT)
● Formalin Ether CT (FECT)
 ● Merthiolate Iodine Formaldehyde CT (MIFCT)

II. FLOTATION
● Uses a soln with a higher specific gravity, w/c causes
parasites to float to the surface. Concentrate can be
removed from the top layer of the tube

A. METHOD
● Zinc Sulfate Centrifugal Flotation Technique (ZSCFT)

● Brine FT
 are easier to perform and less prone to technical errors
- ST used at CDC is formalin-ether acetate technique, a
diphastic ST that avoids the problems of flammability of
ether and can be used with specimens preserved in
formalin, MIF, and SAF
- Suspension of feces in a fluid that’s lighter that the
parasitic forms
- The latter sink to the bottom of the suspension by
gravity
- Expedited by centrifugation

FLOTATION
- Most used is ZSCFT/Sheather’s sugar
- Solution used should have higher specific gravity than
organisms to be floated so that organisms rise to the top
SEDIMENTATION AND FLOTATION and debris sinks to the bottom
- Suspend washed sediments in a soln having slightly
● Direct examination of stool may not alway be able to greater specific gravity that parasite elements so that
reveal the presence of small numbers of parasites when centrifuged, the cysts, eggs, and larvae
● Purpose of CT is to increase the possibility to finding concentrate on the surface film when they can be
protozoan cyst, helminth eggs, and larvae by decreasing removed to a slide of examination, while heavier
the amount of background material elements are thrown to the bottom
● Direct examination should be done first before
proceeding to fecal concentration. Some infections may Advantage: produce cleaner material than sedimentation
be light technique
Disadvantage: walls of eggs and cysts will often collapse, thus
SEDIMENTATION hindering identification, some parasites do not float
- Uses solutions of lower specific gravity than the
parasitic organisms ● Brine Flotation - oldest CT; simple and efficient for
- Recommended for general diagnostic lab because they recovery of all eggs other than operculated and
 Schistosome eggs

PURPOSE OF CT
● Purpose of CT is to increase the possibility to finding
protozoan cyst, helminth eggs, and larvae by decreasing
the amount of background material
● Direct examination should be done first before
proceeding to fecal concentration.
● Some infections may be light
● CT will increase the number of organisms detected
compared with direct microscopy
● Motile protozoan trophozoites are not found in
FORMALIN
concentrated prep
- Combustible
● Concentration procedures and techniques may be
performed on preserved/fresh stool specimen

SAFETY IN THE LAB

● GLOVES should be used when examining stool
specimen
● For FECT, main safety concern would be storage and
handling of formalin and ether

ETHER
- Extremely flammable
- RA 9165, regulation of the sale & storage of ethyl ether

COLLECTION AND EXAMINATION

 Schistosoma haematobium
- Termina urine specimen (last 10-20 mL passed) or a 24
Hour Collection Specimen should be collected in a
sterile container for the dx of Schistosoma haematobium

B. Perianal Swabs
- Used for Enterobius vermicularis or seatworm
- Collected using scotch tape (with its sticky surface
exposed outward) in a tongue depressor whose end is
made flat
- Sticky part is swabbed into anal opening and placed in
the slides

C. Sputum
- Lung flukes Paragonimus westermani
- Larval stage of Ascaris lumbricoides, Stronglyloides
stercoralis and hookworms can be isolated due to lung
migration of nematodes
- Early morning “deep sputum” samples are necessary for
testing and are placed in sterile container w/ proper
labeling

D. Aspirates
- Obtained from diff sites of the body and are collected by
cytopathology staff
SPECIMEN COLLECTION AND PROCESSING OF
- Fine needle aspirates and duodenal aspirate are the most
HUMAN BIOLOGICAL SAMPLES
common types sent to lab
A. Urine
PARASITOLOGICAL SPECIMEN
- Species encountered in U/A is Trichomonas sp. &
 ACCURACY AND RELIABILITY
IMPORTANCE OF DIAGNOSTIC PARASITOLOGY 1. Training and experience of MT/Microscopists
(DP) 2. Available manuals and procedures on standard
● Tropical diseases are seen in non-endemic areas techniques
- Expansion of world travels 3. Instruments and reagents
- Access to more geographic area and population 4. Communication between the lab and clinicians
● Increased no. of immunocompromised patients
- Opportunistic parasites - caused serious disease PARASITOLOGICAL DIAGNOSIS
● Need for individuals who are familiar w/ DP A. Ova/Cyst
B. Trophozoite
DP are based on guidelines of: C. Larvae
1. American Society of Microbiology
2. American Society of Parasitologist METHODS OF DIAGNOSTIC
3. American Society for Medical Technology 1. Microscopy - detection of parasites
4. College of American Pathologists
5. Clinical and Laboratory Standards of Institute 2. Serology-Based Assays
a. Enzyme-linked Immunosorbent Assays -
antigen/antibody detection
 b. Immunofluorescent antibody test
c. Immunoblotting
d. Rapid diagnostic tests (RDTs)

3. Molecular-based assays
a. PCR-nucleic acid-based assay (conventional, real-time,
digital droplet PCR)
b. Loop-mediated Isothermal Amplification
c. Mass spectrometry-Protein markers

FORMS OF PARASITES

MODE OF TRANSMISSION (PARASITIC LIFE CYCLE)

● Soil-borne
- Ascaris lumbricoides
- Trichuris trichiura
- Hookworm
 ● Direct-transmission
- Pediculus
- Enterobius

● Food-borne
- Capillaria
- Paragonimus

● Arthropod-borne
- Plasmodium

● Snail-borne
- Schistosoma

COMMON PROBLEMS IN IDENTIFICATION OF

PARASITES
 ANEMIA AND Major diseases they cause
MALNUTRITION

RESERVOIRS

● They dwell in various habitats and may thrive

either in the environment or require host
throughout life cycle
● Helminth species are mostly found in:

HELMINTHOLOGY Contaminated soil (soil-transmitted), water (blood flukes,

cercariae), food (Taenia spp.)
TERMINOLOGIES Insect Filarial worms
vectors/arthropods
MEDICAL Study of helminths or parasitic
HELMINTHOLOGY worms and how they affect human Wild animals harboring Echinoccocus spp.
health parasite

Considered metazoans or Human hosts Enterobius, Hymenopolis

multicellular organisms

Distributed worldwide and account CLASSIFICATION

for a high frequency of morbidity NEMATODES (ROUNDWORM)
and mortality, especially developing TREMATODES (FLUKES)
countries CESTODES (TAPEWORMS)

NOTE: Trematodes and Cestodes are considered flatworms because

their bodies are flattened and are bilaterally symmetrical impaired learning ability
● MAN is the only host
● Developed in the soil and transmitted via soil
medium either by INGESTION OF
EMBRYONATED EGG or SKIN
PENETRATION BY INFECTED LARVAE

SOIL-TRANSMITTED HELMINTHS
MODES OF TRANSMISSION OF STH
● Most common parasite
● Humans get infected by
● More than 1.5 billion people, or 24% of the world’s
1. Ingesting foods harboring the infective ova
population are affected by STH infection
(uncooked, not thoroughly washed or unpeeled
● STH are transmitted by eggs present in human feces
vegetables)
w/c in turn contaminate soil in areas where
2. Drinking water from contaminated sources
sanitation is poor
3. Children putting their soiled hands in their mouth,
● Intestinal obstruction, anemia, malnutrition,
ingest eggs and become infected
dysentery syndrome, fever, dehydration, vomiting,
4. Skin penetration by Strongyloides or hookworm
colitis
larvaes
● STH infections is prevalent among children due to
poor growth, reduced physical activity, and
HABITATS OF STH IN THE HOST FACTORS AFFECTING TRANSMISSION

SMALL INTESTINE Ascaris lumbricoides 1. Environmental condition

Hookworm ● Soil condition, climate
Strongyloides stercoralis 2. Socio-economic
● Sanitation facilities, water supply, education
3. Health behaviors
LARGE INTESTINE Trichuris trichiura
● Defecation habits
● Personal hygiene
● Occupation-related

INTESTINAL NEMATODES
1. Ascaris lumbricoides
2. Trichuris trichiura
3. hookworm
- Necator americanus
- Ancylostoma duodenale
4. Strongyloides stercoralis
 children
➔ 84% SAC
➔ 30.9% PSAC
➔ 59% PSCA
➔ 54.4% SAC
➔ 35.1 PSCA/SAC

Ascaris lumbricoides
● Giant intestinal roundworm
● Most common intestinal roundworm
● Widely distributed in the Philippines: rural, urban areas MORPHOLOGY
● Common among children; preschool aged, school aged ● Lips:
3 lips - center buccal cavity
Females are larger compared to their
● Male: male counterparts, but males have a
Size: 15-31 cm x 3 mm more curved or coiled tail
Posterior end: curved
Majority of the species are free-living
● Female:
Size: 20-35 x 5 mm
Posterior end: straight
Anterior 1/3: depression/constricted vagina (genital ring)

FACTS

NEMATODES Have unsegmented, elongated, and

(ROUNDWORM) cylindrical bodies. They have a
tough covering called cuticle which
serves as the overall covering of
nematodes and is considered the
outer layer of their bodies

They also possess a middle and

innermost layer called hypodermis
and wall musculature

They have complete digestive tract

equipped with oral and anal opening Ascaris lumbricoides (unfertilized egg)
● Never undergoes further development
Sexes can be separated and identified ● Content of egg is irregular and disorganized
based on size and tall shape ● Larger than fertile egg
 ● Mammillation (cortication) may be irregular or absent
● 88-93/38-40 micrometer
● Has granules, no egg cells

Ascaris lumbricoides (fertilized egg)

● Broadly oval, global brown color
● Single cell stage when passed with feces
● Has 3 layers of egg cells
1. Inner non-permeable, lipoidal, vitelline membrane
2. Thick transparent middle layer or glycogen
membrane
3. Outermost coarsely mamillated, albuminoid layer
LIFE CYCLE OF ASCARIS LUMBRICOIDES - Perforate the bowel - peritonitis
1. Adult worms live in small intestine; a female worm
produces eggs that are passed out with feces DIAGNOSIS
2. Fertilized eggs embryonated after 18 days to several 1. Clinical signs & symptoms
weeks in the soil (infective stage) Vague: confirmatory by lab diagnosis
3. Fertilized eggs are ingested by humans Diagnostic: children-passing out Ascaris
4. Larvae hatch and invade the intestinal mucosa, carried 2. Laboratory
via the portal and then systemic circulation to the lungs Basic Techniques:
and mature in 14 days - Gross examination of the adult stages coming out of
5. Larvae penetrate alveolar walls, ascend to bronchial tree body openings
to the throat and swallowed - Stool examination - microscopic examination for ova
6. Larvae reaches small intestines and develop into adult A. DFS
worm ● Saline
7. 2 and 3 months from ingestion of the infective eggs to ● fresh/preserved stool
oviposition by the adult female B. Concentration techniques
● Kato Katz/Kato Thick
PATHOLOGY AND CLINICAL MANIFESTATIONS ● FECT
1. Reaction to migrating larvae
- Pulmonary manifestations Other/specialized techniques
- Asthmatic-like attacks (bronchial rales and chest pain) ● Direct examination of sputum for larvae
- Pneumonitis - massive Ascaris infection ● Serology for extraintestinal ascariasis
- Loeffler’s syndrome
2. Adults in the duodenum and jejunum TREATMENT
- feed on nutrients ● Individual Infections
3. Wandering adult worm - erratic behavior ➢ Single-dose of broad spectrum anthelmintics
- Entangled - intestinal obstruction - Albendazole, mebendazole, and pyrantel pamoate
- Appendix - acute appendicitis ❖ 400 mg single dose Albendazole
- Bile duct - biliary ascariasis ❖ 500 mg Mebendazole
- Liver parenchyma - multiple abscesses ❖ 10 mg/kg body weight Pyrantel pamoate
- Block the pancreatic duct - acute pancreatitis ➢ Cause paralysis of the parasite
 ➢ Ascaris adults don’t have organs for attachment to the
intestinal ● Female:
➢ 100% cure rate Size: 35-50 mm
Posterior end: blunt
● Community Posterior half: tout
➢ 3x/yr for 3 years
➢ Mass treatment ● Both: Esophagus
➢ Selective treatment: Anterior 1/5: lined w/ stichocytes/stichosomes
- (+) eggs
- Targeted group (children)

PREVENTION AND CONTROL

1. Discriminate fecal disposal - sanitary disposal of feces
2. Personal family - community hygiene
3. Mass treatment - 2-3x/year; children (target
population)

Trichuris trichiura
● Trichuriasis
● Whipworm, whip-like Trichuris trichiura (eggs)
● Habitat: large intestine (cecum) ● 50-54 microns x 22-23 microns
● Barrel or lemon-shaped
MORPHOLOGY
● Lips:
3 lips - center buccal cavity

● Male:
Size: 30-45 mm
Posterior end: coiled
 3. Attached on walls of the large intestine feed on
intestinal tissues, colon, rectum (rectal, anal
prolapse)
4. Prolapse; frequent LBM, loss of muscle tone of the
anal sphincter
5. Invade appendix

DIAGNOSIS
Basic Techniques:
- Stool examination - microscopic examination for ova
C. DFS
● Saline
LIFE CYCLE OF TRICHURIS TRICHIURA
● fresh/preserved stool
D. Concentration techniques
● Kato Katz/Kato Thick
● FECT

Other/specialized techniques
● Examination of the rectal mucosa (by proctoscopy or
directly, in case of prolapses) can occasionally
demonstrate adult worms

EPIDEMIOLOGY
1. Prevalence - 80-84% parallels that of ascariasis
PATHOLOGY AND CLINICAL MANIFESTATIONS 2. Most infections are light to moderate
1. Light infection - no symptoms, insignificant 3. Eggs are susceptible to dryness
2. Heavy; chronic - (>5000 eggs/gm feces) 4. Long-life span of the worm 2 years
- Blood-stroke diarrhea, abdominal pain, nausea, 5. 60 M eggs in 2 years
vomiting, anemia 6. Children; 5-15 y/o
7. Factors affecting transmission as in Ascaris
HOOKWORMS - 4-8 cell stage in the feces
1. Necator americanus - human hookworm - Differentiation of Necator egg from Ancylostoma is
2. Ancylostoma duodenale - human hookworm difficult
3. Ancylostoma ceylanicum - In delayed transit time, embryo may develop inside the
4. Ancylostoma caninum - dog hookworm shell
5. Ancylostoma braziliense - cat and dog - Differentiation of spp. is difficult (human vs. animal
- Adults suck blood; attached to the mucosa of the small spp.)
intestine (jejunum)
Female: 9-13 mm long with egg-filled uterus
CHARACTERISTICS Male: 7-11 mm long, posterior end forms a
1. Necator americanus - semi-lunar cutting plate
2. Ancylostoma braziliense - 2 pairs of teeth NOTE: Ovum of hookworm can develop into two forms:
3. Ancylostoma caninum - 3 pairs a. Rhabditiform Larvae
4. Ancylostoma duodenale - 2 pairs - 250-300 μm
- Long
- 15-20 μm wide
- Long buccal canal, small genital primordium
- Usually not found in stool but may be found if there is delay
in processing
b. Filariform Larvae
- 500-600 μm long
- Pointed tail

MORPHOLOGY
● Eggs
- Ovoidal
- 56-60 x 34-40 micro
 MORPHOLOGY OF LARVAE
Hookworm

RHABDITIFORM FILARIFORM

Short and stout/fat Longer, slender

Short esophagus; muscular Slender and elongated, short

flask (25% body length)
esophagus

Small genital primordium Sheath

Pointed posterior end Pointed posterior end

Feeding stage; long, open Non-feeding stage; closed

buccal capsule

Infective stage
PATHOLOGY AND CLINICAL MANIFESTATIONS
1. At the site of entry larvae
- Ground or dew itch (itching due to penetration of the
filariform larvae, a hypersensitivity rxn because of
enzymes)
- EOSINOPHIL (WBC) ANG TUMATAAS KAPAG
MAY PARASITIC INFECTION!
- Rash around the area of penetration
- Itching, edema, erythema, papulovesicular eruptions
that can last for 2 weeks
2. In the lungs during larval migration
- Bronchitis or pneumonitis: abundant number of
migrating larvae
- Sputum production, cough
3. Small intestine
- Habitat of adult worm
 - Microcytic, hypochromic anemia - chronic moderate
or heavy infection, kind of anemia where there’s a ● Culture Methods:
presence of hookworm B. Harada-Mori
- Iron-deficiency anemia (due to bleeding/blood loss from - Allow hatching of larvae from eggs on filter paper strips
attachment of adult worms to intestinal mucosa with one end immersed in water
● 0.03 - 0.05 ml blood/day - N. americanus - Recommended for species identification
● 0.16 - 0.34 ml - A. duodenale
- Hypoalbuminemia - low level of albumin (blood loss, TREATMENT
lymph, and protein) ➢ Broad spectrum anthelmintics
- Weight loss, abdominal pain - Albendazole
- Mebendazole
PATHOLOGY AND CLINICAL PRESENTATION - Pyrantel pamoate
● Creeping eruptions or cutaneous larva migrans ➢ A severe anemia: raise hemoglobin level
● Intense pruritus - Ferrous sulfate (200 mg 3x/day) - oral for 3 months
● Serpiginous tunnel ➢ Topical antihistamine - CLM
PREVENTION AND CONTROL
1. Proper fecal disposal
2. Use footwear
3. Avoid ingestion of unwashed/improperly washed
vegetables (A. duodenale)
4. Health education, personal, family, community hygiene
4. Treatment of (+) cases to prevent spread

STOOL CULTURE METHOD

● Allows spp. identification of hookworm and
differentiation from Strongyloides by allowing larval
development
DIAGNOSIS
● Stool examination - identification of hookworm ova A. HARADA-MORI/TEST TUBE CM
A. DFS, Kato Katz/Kato Thick, FECT - Smear stool positive for hookworm or strongylid
 ova in microscopy in a filter paper
- Submerge the end of filter paper in a saline
solution carefully not touching the stool smear
- Incubate in a dark room for 3 days
- Centrifuge and examine saline soln after
incubation

Strongyloides Stercoralis
- Threadworm
- Strongyloidiasis
- Tropical and subtropical countries, occur in temperate
zone
- Parasitic and free-living
- Only spp. capable of autoinfection (replicate in the
human host, and may cause infection w/out the need for
repeated exposure), persistent infection

Complications of Strongyloidiasis
- Acute with urticaria, abdominal pains and diarrhea
which may be complicated by malabsorption
- Disseminated w/c occurs in immunocompromised hosts, LIFE CYCLE
usually fatal involving small intestines, lymph nodes, ● Female adults are parthenogenetic, they are able to
lungs and brains produce offspring w/out fertilization w/ males
● Female worms are ovoviviparous, they lay eggs
containing larvae that hatches out immediately

MORPHOLOGY OF LARVAE OF
Strongyloides stercoralis
 RHABDITIFORM FILARIFORM a. Rhabditiform larvae are passed out in the stool w/c
may
Short, open buccal capsule Closed buccal capsule - Molt twice and become infective filariform larvae
(feeding stage) - Molt four times and become free-living adult males and
females that mate and produce eggs from w/c new
Muscular, elongated esophagus Long esophagus (40% body generation of rhabditiform larvae hatch w/c may either
with pyriform posterior bulb length) occupying half the ➔ Develop into new generation of free-living adults
length of the larvae ➔ Develop into filariform larvae
Large genital primordium, Infective stage b. Filariform larvae penetrates human skin to start the
small, conspicuous next cycle
Pointed posterior end Forked/notched tail
2. Parasitic Cycle
A. Filariform larvae are transported to the lungs via
bloodstream of lymphatics were they penetrate alveolar
spaces
B. Larvae are carried through the bronchial tree into the
pharynx
C. Swallowed and reaches the small intestine
D. Molt twice and become female adult worms
E. Live in the intestines and produce eggs by pathogenesis
F. Rhabditiform larvae are hatched
The cycle then repeats or enter autoinfection wherein;
1. Rhabditiform larvae become filariform larvae (mahilig
magpenetrate!)
2. Penetrates either the intestinal mucosa (internal
autoinfection) or skin of perianal area (external AI)
3. Filariform larvae goes to the lungs, bronchial tree,
TYPES OF LIFE CYCLE pharynx, swallowed, and reaches the small intestine
1. Free-living 4. Filariform larvae mature into adults or may disseminate
 into the body. - Thiabendazole
- Ivermectin - chronic uncomplicated strongyloidiasis
PATHOLOGY AND CLINICAL MANIFESTATIONS available ivermectin for humans in the Phil., or animals
1. Acute strongyloidiasis ➢ Prevention and control - as hookworm infection
- Invasion of the skin by filariform larvae
- Migration of larvae thru the body DIAGNOSIS
- Penetration of intestinal mucosa by adult female worms ● Stool examination - Kato Katz/Kato Thick
2. Light infection ● Direct Fecal Examination
- No intestinal symptoms ● Flotac/Mini flotac
3. Moderate infection ● Stool Culture
- Diarrhea alternating with constipation ● Concentration Technique - AECT, FECT
4. Heavy infection
- intractable , painless, intermittent diarrhea characterized Kato Katz
by numerous episodes of watery and bloody stools ● QUANTITATIVE
5. Immunocompromised patients (cancer, malnutrition,
HIV/AIDS)
- Disseminated infection

EPIDEMIOLOGY
1. Found throughout the world, follows distribution pattern
similar to hookworm infection, tropics and subtropics,
as well as EU, US
2. 50-100 million infected
3. In the Phil., 0-2.3%, depending on area selected
4. More frequent in male children, 7-14 y.o than female
and adults
5. Autoinfection FECT
● QUALITATIVE
TREATMENT
- Albendazole
FLOTAC & MINI-FLOTAC
● QUANTITATIVE
HARADA MORI
● Useful for detection of infections with hookworm,
Strongyloides, and Trichostrongylus as well as to
facilitate specific identification of their larval stage
● Fecal material to be cultured shouldn’t be refrigerated
since some spp. will fail to develop following
refrigeration for several hours or longer

STOOL CULTURE METHOD

CONTROL PROGRAMS
● NTD roadmap, published by WHO in 2012, set two
targets for the control STH by 2020
a. 75% of pre-school and school age children in need of
treatment are regularly treated
b. 75% coverage with preventive chemotherapy is achieve
in pre-SAC and SAC in 100% countries

WHO
● Prevent and control morbidity thru the periodic
treatment at risk population
- PSAC
- SAC
- Women of child-bearing age

DOH
● Conducts to prevent the spread of infections of
parasites, etc.
 ● STH control programs ● Caused by Enterobius vermicularis
● Integrated Helminth Control Program ● Human pinworm
● DepEd - school-based MDA ● Perianal itching
● Community-based MDA ● Not fatal; general welfare of the patient merits
attention
INTEGRATED HELMINTH CONTROL PROGRAM
● Occurs worldwide; occur most frequently in school
● Disease burden of STH in the country among vulnerable
or PSA children and in crowded conditions
and high-risk groups id quite high and way above the
global standard of less than 20% cumulative prevalence
● Reduce the CP of STH to less than 20% and prevalence
of Moderate to Heavy Intensity Infection (MHII) to less
than 2%

OTHER CONTROL PROGRAM

1. WASH - Water, sanitation, and hygiene

A. Adult
B. Egg
C. Adult male and female
D. Adult male and female
E. Male

DIRECTLY-TRANSMITTED PARASITES MORPHOLOGY OF ENTEROBIUS

VERMICULARIS EGG
ENTEROBIASIS
● Size ranges from 50-60 micrometer by 20-30
● Asymmetrical with 1 side flattened and the other
convex
● With translucent shell
DIAGNOSIS
● Basic techniques
- Microscopic examination of perianal swab
❖ Collection is ideally done in the morning before
defecation and washing
❖ Done by pressing a transparent adhesive tape on the
perianal skin and examining the tape placed on a
 slide
● Eggs
❖ Can also be found, but less frequently in the stool,
and occasionally encountered in the urine or vaginal
smears
● Adult
❖ Diagnostic, when found in the perianal area or
during ano-rectal or vaginal examinations
● Finding the eggs or adult worms via microscopy
● Graham’s scotch adhesive tape swab (perianal
cellulose tape swab)

TRICHOMONIASIS
● Caused by the parasite Trichomonas vaginalis
● Sexually-transmitted
● Correlates strongly with the number of sexual partners
● MALE PARTNER ANG CARRIER PERO SA
FEMALE NAKIKITA
● Humans are the only natural host
NOTES:
● Trophozoite - infective stage, nakukuha thru sex; oral or
vaginal
● pH - 3.8 to 4.2
● Pathogenic is in the genitourinary tract
● Pus - green/yellow
- Gram Staining