Parasitology (Lecture)

Why Study Parasitology?  Thorny- headed worms

- To monitor African countries: Haiti, Guatamela, - Infects animals
Myanmar - Rarely infect humans
- Undeveloped tropical and subtropical countries with
high population density, poor sanitation, marginal water Needed for Collection
sources, poor public health practices, environmental 1. Sterile screw- cup container- clean & water tight
changes affecting vector breeding areas. 2. Pea- sized stool
3. Not from toilet bowl/ diaper
Parasitic Life Cycle
- Is one phase involve the route of parasites follows when YES!! (Preservatives and Fixation)
in or on the human body.
- The other phase, the route a parasite follows Formalin
independently of the human body, provides crucial - 5%- protozoan cysts
information pertinent to epidemiology, prevention and - 10%- helminths egg
control. Advantages:
- Three common components--- 1. Easily prepare
 A mode of transmission 2. Preserve up to years
 Infective stage 3. Long shelf- life
 Diagnostic stage
 Some parasites require only a definitive Disadvantages
host, whereas others also require one or 1. Not preserved morphology for smears
more intermediate hosts.
Polyvinyl Alcohol (PVA)
What can be seen in Stool? - Preparing slides
 Ova - Combined Schaudinn solution w/c contains
 Parasite zinc sulfate, copper sulfate or mercuric
chloride as base
Specimens
 Stool Na+ Acetate Formalin/ Sodium Acetate Formalin (SAF)
- Used performing technique and permanent
 Blood
stains
 Tissue biopsies
- Only requires single vials without mercury/
 Sputum
mercury- free
 Urine Advantages:
 Genitals 1. Easy prepare
 CSF- cerebrospinal fluid; clear, colorless that flow 2. Long shelf life
around brain and spinal cord. 3. Used for preparing smears for staining modified Acid
Fast Bacilli Stains (AFB) to detect coccidian oocysts
Parasite Disadvantages:
 Genus 1. Need an addition of albumin to ensure adhesion of spx
 Species to slides.

Ex. Modified PVA

Genus Species - Used substitute compound containing
Ascaris Lumcoides copper sulfate or zinc sulfate
A. Lumbricoides - Permanent stained smears and concentration
method
Collection and Transport Advantages:
1. Properly collected and prepared stool specimen. The 1. Free form formalin and mercury
protozoan forms known as trophozoites and cysts may 2. Used for concentration technique and permanent stain
be recovered from these samples. 3. Used for fecal immunoassays
2. Helminths stages, such as eggs, larvae, proglottids, and
adult worms, may also be found. Definitions of these Disadvantages
helminth—related morphologic forms are detailed in 1. Do not provide same quality of preservation as
the corresponding parasites chapters of this text mercury based fixatures organisms ID will be more
3. The typical stool collections protocol consists of three difficult from permanent stain.
specimens, one specimen collected every other day or a
total of three collected in 10 days. One exception is the Processing
diagnosis of amoebiasis, in which up to six specimens Consistency Possible Gross Appearance
in 14 days is acceptable. color

Helminths 1.Hard Dark brown Conspicuously fibrous

 Roundworm- infects intestine
 Flukes- infects = bile ducts, liver/ blood
2.Soft Black Fiber scanty
 Tapeworms- infects intestine

Veraye, Angela O.
Parasitology (Lecture)

3.Mushy Brown Colloidal - The advantage of this technique is that it provided good
(homogenous) recovery of most parasites and is easy to perform
4.Loose Pale brown Scanty mucus - The disadvantage of this technique is that the
preparation contains more fecal debris than a floatation
technique and is more challenging to the microscopist.
5.Diarrheic Clay Much mucus
6.Watery/Liquid Yellow Mucus scanty blood Zinc Sulfate Floatation Technique
- Zinc sulfate with a specific gravity of 1.18 to 1.20, is
used as the concentrating solution
7.Formed Red- brown Others…. - When zinc sulfate is added to the specimen and
centrifuged, the parasite float

8.Semi-formed Green, others Permanent Stains

1. Wheatley Trichrome- most widely used permanent stain
 Uses reagent with a relatively long shelf life
Soft & Water/ Liquid and the procedure is easy to perform
- Protozoan cysts/ trophozoites 2. Iron Hematoxylin- may be used instead of the trichrome
- Helminths (egg/ larvae) technique to be time- consuming
 Reveals excellent morphology of the
30 min- liquid intestinal protozoa
1hr- formed 3. Specialized Stains- disadvantage of these stains is that
they do not detect oocysts of the coccidian parasites or
Microscopic Examination spores of microsporidia. Example is modified Acid Fast
- The microscopic examination of stool for ova and Stain
parasites involves three distinct procedures
 Direct wet preparations Stools Screening Methods
- These techniques allow detection of a wide variety of
 Concentrated technique
parasites
 Permanently stained smear
- Are labor- intensive and require an experienced
microscopist. Alternative tests have been developed that
Direct Saline Wet Preparations
are often referred to as rapid methods, or stool
- Made by placing a drop of 0.85% saline on a glass slide
screening methods. These methods can be obtained as
(a 3-x- 2- inch size is suggested) and mixing with a
kits that contain monoclonal antibody.
small portion of unfixed stool using a wooden
- Include enzyme immunoassay (EIA), direct fluorescent
applicator stick or another mixing tool.
antibody (DFA), and membrane flow cartidge
techniques.
Direct Iodine Wet Preparation
- Made to enhance the detail of protozoan cysts
The Nematodes
- Use drop of iodine (Lugol’s or D’ Antoni’s formula) in
place of saline
Morphology and Life Cycle Notes
- Iodine kills any trophozoites present
- Members of the class Nematodes (multicellular
- Recommended to use direct saline and direct iodine wet
parasites that appear round in cross section)
preparations on each sample that requires this
- Three basic morphologic forms: eggs (female sex cells
component of testing
after fertilization), juvenile worms known as larvae, and
adult worms
Direct Wet Preparation (also known as direct wet mount)
- Sexes are separate. The adult female worms are usually
- Primary purpose, defined as a slide made by
larger that the adult males.
trophozoites present
- Fertilized adult female nematodes lay their eggs in the
intestine. These eggs may be passed into the stool
Concentration methods
- Once outside the body, the larvae located inside the
- Provide the ability to detect small numbers of parasites
eggs require warm moist soil and 2 to 4 weeks for
that might not be detected using direct wet preparations
continued development. The developed egg is then
- The purpose of concentration is to aggregate parasites
ready for infection into a new host and the cycle is
prenset into a small volume of the sample and to
repeated.
remove as much debris as possible that might hinder the
lab technician
Laboratory Diagnosis
- May be accomplished by the recovery of eggs, larvae
Formalin- Ethyl Acetate Sedimentation Procedure
and occasional adult worms
- The principle of this technique is based on specific
- The specimens of choice vary by species and include
gravity
cellophane tape preparations taken around the anal
- Ethy acetate is added to a saline- washed formalin-
opening, stool samples, tissue biopsies and infected skin
fixed sample and the tube is then centrifuged
ulcers.
- Parasites are heavier than the solution whereas fecal
debris is usually lighter and rises to the upper layers of
Enterobius vermicularis
the test tubes.
Common name: pinworm, seatworm

Veraye, Angela O.
Parasitology (Lecture)

Common associated disease: enterobiasis, pinworm infection infective eggs may then become dislodged from the
body, caused at least in part by intense scratching of the
Parameter Description anal area by the infected person. Once apart from the
Size 48-60µm long, 20-35 µm host, the infective eggs may take up residence in a
number of locations, including dust, sandboxes, linens,
and clothing. In addition, the eggs may become
Shape Oval, one-side flattened airborne. The infective eggs may survive for a few days
up to several weeks

Embryo Stage of development varies; may

be unembryonated, embryonated, Epidemiology
mature - E. vermicularis is found worldwide, particularly
intemperate areas
- Pinworms is considered by many to be the most
common helminth known to cause infection in United
States.
- E. vermicularis may be responsible for the transmission
Shell double layered, thick, colorless of Dientamoeba fragilis. This theory suggests that the
D. fragilis trophozoite may actually take up residence
inside the pinworm egg for transmission

Characteristics Adult Females Adult Males Clinical Symptoms

- Asymptomatic: many cases of E. vermicularis infection
are asymptomatic
Length 7-14 mm 2-4mm - Enterobiasis: Pinworm infection: most common
symptoms experienced by the individuals infected with
Width Up to 0.5 mm <0.3mm pinworm include intense itching and inflammation of
the anal/ or vaginal areas. These symptoms may be
Color Yellowish-white Yellowish- white accompanied by intestinal irritation, mild nausea or
vomiting, irritability, and difficulty sleeping.
Tail Pointed;
Resembles
Pinhead Trichuris Trichiura
*Adult males are only rarely seen Common Name: Whip worm
Common Associated disease and condition names: Trichuriasis,
Unfertilized egg- unembryonated egg whipworm infection
Fertilized egg- embryonated egg
Parameter Description
Retroinfection- defined in pinworm- specific terms as infective Size 50-55 by 25 µm
pinworms that migrate back into the host body Shape Barrel, football; hyaline polar
Autoreinfection- if infective pinworms eggs are ingested via hand- plug at each end
Embryo Unicellular; undeveloped
to- mouth contamination
Shell Smooth; yelloe- brown color
because of bile contact
Laboratory Diagnosis
- Choice for the recovery of E. vermicularis is a
cellophane tape preparation collected from the perianal
Laboratory Diagnosis
region of the person suspected of infection
- The specimen of choice for the recovery of T. Trichiura
- Although eggs are the primary morphologic form seen,
eggs is stool
adult females may also be present if the sample is
collected when the females enter the perianal region to
Life Cycle
lay their eggs
- Ingestion of infective T. trichiura eggs containing
larvae initiates human infection. The larvae emerge
Life Cycle
from the eggs in the small intestine.
- Humans are the only known host of E. vermicularis.
Growth and development of the larvae occur as they
- Pinworm infection which is usually selflimiting
migrate within the intestinal villi. The larvae return to
- the ingestion of the infective eggs. The eggs migrate
the intestinal lumen and proceed to
through the digestive tract into the small intestine,
the cecum, where they complete their maturation. The
where they hatch and release young larvae. The
resulting adults take up residence in the colon,
resulting larvae continue to grow and mature, ultimately
embedding in the mucosa. The life span of the adult
transforming into adult worms. The adult worms reside
worms in untreated infections may be from 4 to 8 years.
in the colon. Following mating of select worms
- • Following copulation, the adult female lays her
(copulation), including roundworms, the resulting
undeveloped eggs. It is this stage of egg that is passed
pregnant (gravid) female worm migrates outside the
into the outside environment via the
body to the perianal region, where she may deposit up
to 15,000 eggs. Following 4 to 6 hours incubation, the
developing eggs achieve infective status. These

Veraye, Angela O.
Parasitology (Lecture)

feces. Following approximately 1 month outside the - • The first stop on this journey is the liver. From there,
human body, usually in the soil, the eggs embryonate, the larvae continue via the bloodstream to the second
become infective, and are ready to initiate a new cycle. stop, the lung.
- • Once inside the lung, the larvae burrow their way
Epidemiology through the capillaries into the alveoli. Migration into
- found primarily in warm climates of the world where the bronchioles then follows.
poor sanitation practices are common, such as
defecating directly into the soil or using human feces as Hookworms: Necator Americanus
a fertilizer. Common Name: New World Hookworm

Clinical Symptoms Hookworm: Ancylostoma duodenale

- Asymptomatic: patients who suffer from a slight Common name: Old World Hookworm
whipworm infection often remain asymptomatic Common associated disease and condition names: Hookworm
Trichuriasis: Whipworm Infection: Heavy infection of infection, ancylostomiasis, necatoriasis
500 to 5000 worms produce a wide variety of Anccylostoma size range: 50-60mm
symptoms
- Children infected with T. trichiura usually present with Differentiation
symptoms resembling those ulcerative colitis - In humans, larvae of Necator must penetrate through
- An infection of as few as 200 worms may cause a child the skin, whereas Ancylostoma infections may be
to develop chronic dysentery, a severe anemia, and percutaneous, oral, transmammary, and probably
possibly growth retardation. It is interesting to note that transplacental
in treated children, catch- up growth usually occurs. In - Ancyclostoma caninum, a hookworm of dogs, the only
addition, increased rectal prolapse and peristalsis are hookworm of Europe and the aeas bordering the
common in infected children. Mediterranean, of the west coast of South America, and
of parts of India and China.
Prevention and Control - Necator americanus, while prevalent over large portion
- Proper sanitation practices, especially avoidance of of the Western Hemisphere was probably introduced
defecating directly into the soil, using feces as a from Africa with the slave trade, the only hookworm
fertilizer, and placing potentially infective hands into found in North America and the large areas of South
the mouth and prompt and thorough treatment of America, it is the native hookworm of Africa, South of
infected persons, when indicated. the Sahara and the only one found in parts of India
- Ancylostoma and necator occur togetherelsewhere in
Ascaris Lumbricoides India, in much of China, in sputheast Asia, in Indonesia,
Common names: Large intestinal roundworm, roundworm of man in the islands of the South and Southwest pacific, and in
Common associated disease and condition names: Ascariasis, part of Australia
roundworm infection - Eggs of necator closely resemble those of
Ancyclostoma but are slightly larger averaging 64 to
Parameter Description 76µm by 36 to 40µm
Size 85-95µm by 38-45µm; size - Adult Ancyclostoma being larger than Necator, worm
variations possible for worm it is more likely to bring about iron deficiency
Shape Varies
Embryo Unembbryonated; amorphous Rhabtiform Larvae
mass of protoplasm - Actively feeding larva will, at a minimum, double in
Shell Thin length, ranging from 540 to 700µm, when it is only 5
Other features Usually corticated days old
- This morphologic form is characterized by the presence
Laboratory Diagnosis of a long oral cavity known as a buccal cavity or buccal
- The specimen of choice for the recovery of A. capsule and a small genital primordium (i.e., a
lumbricoides eggs is stool) Adult worms may be precursor structure to a reproductive system consisting
recovered in several specimen types, depending on the of a clump of cells in an ovoid formation)
severity of infection, including the small intestine,
gallbladder, liver, and appendix. • In addition, adult Filarifrom Larvae
worms may be present in the stool, vomited up, or - The infective, nonfeeding filariform larva emerges after
removed from the external nares, where they may the rhabditiform larva completes its second molt.
attempt to escape. An enzyme-linked immunosorbent - First, this slender larva has a shorter esophagus than
assay (ELISA) is also available. that of Strongyloides stercoralis, a similar intestinal
nematode (covered later in this chapter). Secondly, the
Life Cycle hookworm filariform larva has a distinct pointed tail.
- • Infection begins following the ingestion of infected Hookworm Filariform Larva: Typical Characteristics at a
eggs that contain viable larvae. Glance
- • Once inside the small intestine, the larvae emerge Parameter Description
from the eggs. The larvae then complete a liver-lung Length of Esophagus Short
migration by first entering the blood via penetration Tail Pointed
through the intestinal wall.

Veraye, Angela O.
Parasitology (Lecture)

- larvae in the stools

Life Cycle - With severe diarrhea, the embryonated eggs may be
- Humans contract hookworm when third-stage filariform present in the stools and can be differentiated from
larvae penetrate through the skin, particularly into areas hookworm eggs by the fact that they always contain
such as unprotected feet. S well-developed larvae
- Lesions resembling the ground itch of hookworm
- Once inside the body, the filariform larvae migrate to infection are sometimes seen following penetration of
the lymphatics and blood system. the skin by Strongyloides larvae
- Pneumonitis may be produced by the larvae, but as in
- The blood carries the larvae to the lungs, where they hookworm infection it is generally less severe than in
penetrate the capillaries and enter A the alveoli. ascariasis
- malabsorption syndrome with steatorrhea
- Migration of the larvae continues into the bronchioles, - Heavy infection may involve the large as well as the
where they are coughed up to the 0 pharynx, small bowel, and ulceration of the intestinal mucosa
subsequently swallowed, and deposited into the may give rise to symptoms suggestive of duodenal ulcer
intestine. or of ulcerative colitis
- Melena may be seen, or there may be massive lower
- Maturation of the larvae into adult hookworms occurs gastrointestinal bleeding, with passage of bright red
in the intestine. blood per rectum

- The resulting adults live and multiply in the small Other

intestine. Adult females lay 10,000 to 20, 000 eggs/day.  Strongyloides Fulleborni- parasites of monkeys, also
infects human, and is common in infants under 6
- Many of the resulting eggs are passed into the outside months of age
environment via the feces. Within 24 to 48 hours and  S. fulleborni kellyi- most infant and adults
under appropriate conditions- warm, moist soil-first- are asymptomatic, but some infants suffer
stage rhabditiform larvae emerge from the eggs. The from what is locally referred to as “swollen
larvae continue to develop by molting twice. Third- belly sickness” characterized by abdominal
stage infective filariform larvae result and are ready to distention, respiratory distress, generalized
begin a new cycle. edema, and hypoproteinemia
 Zoonotic Strongyloidiasis/ Creeping Eruption- bearing
- A number of species of Strongloides are known, and an interesting relationship to “swimmer’s itch” caused
generally they are quite host specific. Cats and dogs by penetration of the skin by avian schistosome
may both be infected with S. stercoralis, and human cercariae
infection from a canine source has been reported.  S. Myopotami of nutria and S. procyonis of
raccoons, and larvae of both are abundant in
swamp waters
Life Cycle
•Life cycle rhabditiform-tarvae in the threadworm are usually Capillaria Philippinensis
passed in the feces. Eggs - first observed in 1962, in the province of locos Sur
are only occasionally found in such samples. - Adult worms are slender, approximately 4 to 5 mm long
• These rhabditiform larvae develop directly into the third-stage - live in the intestinal mucosa, primarily in the jejunum,
infective filariform where they may be present in enormous numbers
larvae in warm, moist soil. - Eggs voided by infected persons, measuring about 45
• The remaining phases of the threadworm life cycle basically by 21 um, develop outside the host and are ingested by
mimic those of the freshwater and brackish-water fish, in which larval
hookworm life cycle. In the indirect cycle, threadworm stages have been found
rhabditiform larvae are passed - Laboratory diagnosis is made by finding the
into the outside environment (soil) and mature into free-living characteristic eggs in the Stool
adults that are - asymptomatic or mildly symptomatic cases occur
nonparasitic. These adult females produce eggs that develop into - Abdominal pain,borborygmus (gurgling), and diarrhea
rhabditiform larvae. appear early
These larvae mature and transform into the filariform stage, at
which time they may
initiate a new indirect cycle or become the infective stage for a
human host and begin
a direct cycle.
• Autoinfection occurs when the rhabditiform larvae develop into
the filariform stage
inside the intestine of the human host. The resulting infective
larvae may then enter
the lymphatic system or bloodstream and initiate a new cycle of
infection.

Diagnosis and Symptoms

Veraye, Angela O.
Parasitology (Lecture)

Order Pseudophyllidea
Order Cyclophyllidea

ORDER PSUDOPHYLLIDEA
1. Dibothriocephalus latus
- Formerly diphyllobothrium latum
- “broadfish tapeworm”
- Insufficiently cooked freshwater fish

LIFE CYCLE:

Eggs are passed unembryonated in feces   . Under appropriate

conditions, the eggs mature (approximately 18 to 20 days)   

and yield oncospheres which develop into a coracidia   . After

ingestion by a suitable crustacean (first intermediate host) the

coracidia develop into procercoid larvae   . Procercoid larvae

are released from the crustacean upon predation by the second
intermediate host (usually a small fish) and migrate into the deeper
tissues where they develop into a plerocercoid larvae (spargana),

which is the infectious stage for the definitive host   . Because

humans do not generally eat these small fish species raw, the
second intermediate host probably does not represent an important
source of human infection. However, these small second
intermediate hosts can be eaten by larger predator species that then

serve as paratenic hosts   . In this case, the plerocercoid

migrates to the musculature of the larger predator fish; humans
(and other definitive host species) acquire the parasite via

consumption of undercooked paratenic host fish   . In the

definitive host, the plerocercoid develops into adult tapeworms in
the small intestine. Adult diphyllobothriids attach to the intestinal
CHAPTER 7: Phylum Platyhelminthes: Class Cestoda
mucosa by means of two bilateral groves (bothria) of their
Cestodes
- Or tapeworms, constitute the Class Cestoda scolex   . The adults can reach more than 10 m in length, with
more than 3,000 proglottids. Immature eggs are discharged from
Adult tapeworm found in human all found flat and ribbonlike body the proglottids (up to 1,000,000 eggs per day per worm) and are
and are white or yellowish passed in the feces. Eggs appear in the feces 5 to 6 weeks after
infection.
2 ORDERS OF TAPEWORMS

Veraye, Angela O.
Parasitology (Lecture)

2. Spirometra spp.
- Infection of human by plecercoid larvae
- Spirometra is known as sparganosis

Veraye, Angela O.