Chapter 6

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Part I

General Microbiology

CHAPTER 6
BASIC CONCEPTS OF
MEDICAL
PARASITOLOGY

Universities Press
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Hyderabad 500 029 (A.P.), India
Phone: 040-2766 5446/5447
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Dr Sonal Saxena, MD
Director Professor and Head of the Department of Microbiology
Maulana Azad Medical College,
New Delhi
and
Dr Amala A Andrews, MD
Maulana Azad Medical College,
New Delhi

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Parasites are eukaryotic organisms with a true nucleus
GENERAL Unicellular: These can be visualised with a microscope,
CONCEPTS OF e.g., protozoa such as Entamoeba histolytica

PARASITOLOGY Multicellular: These can be seen with the naked eye, e.g.,
helminths such as roundworms

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Infective forms: Specific stages of the parasite life cycle which can
establish infection

LIFE CYCLE OF Habitat: A place in which the parasite naturally lives and grows
PARASITES Host: An entity that provides nutrition and harbours the parasite
◦ Definitive host: This is a host in which the adult stage of the parasite
lives/undergoes sexual reproduction
◦ Intermediate host: A host that harbours the larval stage of the
parasite/in which asexual reproduction takes place
◦ Reservoir host: Harbours the parasite and also acts as a source of
infection
◦ Paratenic host: The parasite is unable to develop further in this host;
it can act as a transport or carrier host

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Ectoparasites: Parasites that live on the external body surfaces
of the host

Endoparasites: These live inside the host’s body


TYPES OF Obligate parasites: These organisms cannot exist without a host
PARASITES Facultative parasites: These organisms can exist in the free-
living state or as parasites

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HOST–PARASITE RELATIONSHIP
Symbiosis: Parasite and host are mutually dependent on each other
Commensalism: Parasite benefits while the host is neither benefitted nor harmed
Parasitism: Parasites derive the benefits of nutrition and shelter from the host they live in; in the
process, the host is harmed

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LIFE CYCLE
OF
PARASITES
R O U T E S O F E N T RY O F
PA R A S I T E S

Fig. 6.1 Routes of entry of disease-causing parasites


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CLASSIFICATION OF PARASITES
Three kingdoms:

(i) Protozoa

(ii) Stramenopila

(iii) Animalia

Protozoa are unicellular, eukaryotic, simple, microscopic parasites

Stramenopila are unicellular, plant-like animals


Helminths are multicellular worms belonging to kingdom Animalia

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PROTOZOOL
OGY
Table 6.1 Common protozoa associated
with human infections and their
characteristics

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HELMITHOLOGY
Table 6.2 Phylogeny of helminths

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HELMITHOLOGY
Table 6.3 Common helminths
and the diseases they cause

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GENERAL CHARACTERISTICS, LIFE CYCLE AND MODES OF
TRANSMISSION OF PARASITES
CHARACTERISTICS OF NEMATODES
Long, cylindrical, bilaterally symmetrical and

Unsegmented, with both ends pointed and without


any appendages

Sexes are separate


Alimentary canal well-developed

Fig. 6.2 Internal structure of nematodes (Source: Textbook of Medical


Parasitology, Dr Sumeeta Khurana and Dr Abhishek Mewara)

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GENERAL CHARACTERISTICS, LIFE CYCLE AND MODES OF
TRANSMISSION OF PARASITES
Intestinal nematodes complete their life cycle in a single host

Nematodes either lay


◦ Eggs (oviparous, e.g., Ascaris), or
◦ Larvae (viviparous, e.g., Wuchereria) or
◦ Eggs that immediately hatch into larvae (ovoviviparous, e.g., Strongyloides)

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GENERAL
CHARACTERISTICS,
LIFE CYCLE AND
MODES OF
TRANSMISSION OF
PARASITES
LIFE CYCLE OF NEMATODES

Fig. 6.3 Schematic representation of the life cycle of nematodes

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GENERAL CHARACTERISTICS,
LIFE CYCLE AND MODES OF
TRANSMISSION OF PARASITES
CHARACTERISTICS OF CESTODES
Segmented, long and flattened dorsoventrally: ‘Tapeworms’

Have a head (scolex), a neck and body (strobila)


Lacks alimentary canal
Sexes are not separate
Gravid proglottids containing hundreds of eggs are released
Fig. 6.5 Schematic representation of the life cycle of cestodes
into the environment

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GENERAL CHARACTERISTICS, Fig. 6.4 Morphology of cestodes (Source: Textbook of
LIFE CYCLE AND MODES OF Medical Parasitology, Dr Sumeeta Khurana and Dr
TRANSMISSION OF PARASITES Abhishek Mewara)

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GENERAL
CHARACTERISTICS, LIFE
CYCLE AND MODES OF
TRANSMISSION OF
PARASITES
CHARACTERISTICS OF TREMATODES
Leaf-like, flat, unsegmented worms
Eggs are oval to round, operculated and
embryonated when laid

Sexes are not separate except in


schistosomes
Several larval forms are seen
Alimentary tract consists of a blind,
bifurcated intestinal tract without an
anus

Fig. 6.6 Morphology of trematodes (Source: Textbook of Medical


Parasitology, Dr Sumeeta Khurana and Dr Abhishek Mewara

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Fig. 6.7 Schematic diagram of the life
cycle of trematodes

GENERAL CHARACTERISTICS, LIFE CYCLE AND


MODES OF TRANSMISSION OF PARASITES

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GENERAL
CHARACTERISTI
CS, LIFE CYCLE
AND MODES OF
TRANSMISSION
OF PARASITES
TA B L E 6 . 4 M O D E S O F
TRANSMISSION OF COMMON
PA R A S I T E S I N F E C T I N G
HUMANS

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PATHOGENESIS Physical obstruction, irritation, and trauma
OF PARASITIC Vascular or lymphatic obstruction: Elephantiasis
DISEASES Enzymatic damage
Allergic reactions
Inflammation
Neoplastic reactions: Schistosoma are
associated with carcinoma

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IMMUNITY IN PARASITIC DISEASES
Cytotoxic T cell response, natural killer cells, activated macrophages and antibodies

Immunological responses against parasites are lower than those against bacteria and viruses due to
the intracellular location of parasites and immunological tolerance

Large and complex antigenic structures and multiple antigenic variations

Premunition immunity: immunity which lasts only while the original infection is active

LABORATORY DIAGNOSIS

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LABORATORY
DIAGNOSIS OF Table 6.5 Recommended samples to demonstrate
commonly encountered parasites
PARASITIC DISEASES
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LABORATORY DIAGNOSIS OF PARASITIC DISEASES
EXAMINATION OF FECAL SAMPLES
Collection of samples
i) The NIH swab is used to collect samples when there is a suspicion
of Enterobius infection

ii) Cellophane tape is applied to the perianal area of the patient


early in the morning

iii) String test for duodenal samples: examined for trophozoites

Fig. 6.8 NIH swab to collect sample for demonstrating eggs of E.


vermicularis (Source: Textbook of Medical Parasitology, Dr
Sumeeta Khurana and Dr Abhishek Mewara)

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Stool concentration techniques
Used to separate parasites from fecal matter
LABORATORY i) Flotation method: Stool sample mixed with solutions
DIAGNOSIS OF with a higher specific gravity so the ova rise to the top

PARASITIC ii) Saturated salt technique: Saturated salt solution added

DISEASES iii) Zinc sulfate floatation: 33% zinc sulfate solution is used
iv) Formol ether sedimentation method: Ether and 10%
solution of formalin are used

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LABORATORY DIAGNOSIS OF PARASITIC DISEASES
Wet mount
To identify protozoan trophozoites, cysts, oocysts, helminth eggs and larval forms
Other staining techniques used to demonstrate ova and cysts
1. Modified acid-fast staining
Identification of oocysts of Cryptosporidium, Cystoisospora, Cyclospora
Chromotrope staining: Used for microsporidial spores

Table 6.6 Stool findings commonly seen in infected patients

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LABORATORY DIAGNOSIS OF PARASITIC DISEASES
2. Permanent staining
To visualise coccidian parasites, trophozoites and ova of protozoa and eggs and larvae of the
helminth

3. Trichrome staining
Simple and rapid method

Used to visualise intestinal protozoa, human cells, yeasts and artifact material in the stool

4. Iron hematoxylin staining

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LABORATORY DIAGNOSIS OF PARASITIC DISEASES
Preservation of stool samples
 If the samples cannot be examined within a few hours
 If the specimen for molecular diagnosis must be shipped to a referral laboratory

•Formalin 10% aqueous


•PVA (polyvinyl alcohol)
•Other fixatives: Merthiolate iodine-formalin (MIF); sodium acetate-acetic acid formalin (SAF)

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EXAMINATION OF BLOOD SAMPLES

LABORATORY Peripheral blood smear (PBS)


 Plasmodium in red blood cells
DIAGNOSIS  Amastigote forms of hemoflagellates (e.g., Leishmania

OF PARASITIC donovani) inside monocytes

 Microfilariae of W. bancrofti and B. malayi


DISEASES  DEC/hetrazan provocation test: to stimulate nocturnal
microfilariae

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LABORATORY DIAGNOSIS OF PARASITIC DISEASES
Blood smears
 Finger-prick method or from blood collected in EDTA tubes

 Thick and thin films made

Thick film

 Screening tool in malaria

 Used to visualise Leishman–Donovan (LD) bodies in leishmaniasis

 To detect microfilariae in filariasis


 Smear from a big drop of blood, dehemoglobinisation done

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LABORATORY DIAGNOSIS OF PARASITIC DISEASES
Thin film
Smear should be one cell thick, fixed with methanol before staining
Staining of blood smear
 Leishman stain
 Giemsa stain
 Wright stain

Fig. 6.9 Thick and thin films (Source: PHIL, Image ID 5905/ CDC)

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LABORATORY DIAGNOSIS OF PARASITIC
DISEASES
Culturing
 Requires special media
 There are three types of culture systems
i) Xenic cultivation: The parasite is grown in the presence of an undefined flora

ii) Monoxenic cultivation: The parasite is grown in the presence of a single species of associate
flora

iii) Axenic cultivation: The parasite is grown in the absence of any other metabolising cells

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LABORATORY DIAGNOSIS OF PARASITIC DISEASES
Diamond medium—for the cultivation of Trichomonas vaginalis

NNN (Novey, Nicoll and McNeal) medium—for Leishmania species

RPMI 1640 with human RBC in 7% carbon dioxide and 1–5% oxygen—used to
cultivate P. falciparum

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LABORATORY DIAGNOSIS OF PARASITIC DISEASES
Antigen detection
 ELISA-based antigen detection kits: Giardia lamblia,
Cryptosporidium species in stool samples and T. vaginalis in
urogenital specimens

 Serological diagnosis: ELISA, immunochromatographic tests


 Skin hypersensitivity tests: E.g., performed to diagnose
hydatid disease (Casoni test)

 Molecular diagnostic method: To diagnose malaria, filariasis


and trypanosomiasis
Fig. 6.10 Rapid antigen detection test for malaria
parasite: (a) positive for Plasmodium vivax and (b)
negative test (Source: Dept. of Microbiology, ([a]
MAMMC and [b] PIMS)

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