MINISTRY OF PUBLIC HEALTH OF UKRAINE

BUKOVINIAN STATE MEDICAL UNIVERSITY

DEPARTMENT OF MEDICAL BIOLOGY,

GENETICS AND HISTOLOGY

V. Pishak, M. Grytsiuk, R. Bulyk

MEDICAL BIOLOGY III.

MEDICAL PARASITOLOGY
Manual for foreign students

CHERNIVTSI - 2006

164 1
УДК: 61:57

Резензенти: Г.І. Ходоровський, доктор медичних наук, професор

кафедри фізіології Буковинського державного медичного
університету;
Н.І.Войткевич, старший викладач кафедри іноземних
мов Буковинського державного медичного університету

The manual is designated for teaching of foreign students in Medical Higher

Educational Institutions of Ukraine with ²²² – ²V level of accreditation

Medical biology III. Medical parasitology: Manual for foreign students / V.Pishak,
M.Grytsiuk, R.Bulyk. – Chernivtsi: Bukovinian State Medical University, 2006. – p.

This manual was formed according to the modern program of medical biology.
The main questions of medical parasitology, protozoology and helmintology are
described here as well as medical entomology, life cycles of different parasites and
types of hosts.
This manual will be useful for future studing of infectious diseases.
We tried to make a compilation from different sources, such as modern textbooks,
journals, atlases and Internet data.
This manual is designated for teaching of foreign students.

© V. Pishak, M. Grytsiuk, R. Bulyk, 2006

© Медуніверситет, 2006
2 163
TOPIC: INSECTA, DIPTERA ..................................................................................... 131
Insects .............................................................................................................................. 131
Cockroaches .................................................................................................................... 134
Heteroptera ..................................................................................................................... 135
Bedbugs ........................................................................................................................... 136
Anoplura .......................................................................................................................... 137
Head lice .......................................................................................................................... 137
Crab Lice ......................................................................................................................... 139
Fleas ................................................................................................................................. 139
Diptera ............................................................................................................................. 141
Mosquitoes ....................................................................................................................... 141
Mosquitoes ....................................................................................................................... 142
Aedes mosquitoes ............................................................................................................ 146
Culex mosquitoes ............................................................................................................ 147
Phlebotomus .................................................................................................................... 147
Black Fly .......................................................................................................................... 148
Horse-Flies ....................................................................................................................... 148
Musca domestica ............................................................................................................. 150
Tsetse ................................................................................................................................ 152
Oestridae .......................................................................................................................... 158
Deer Botfly ...................................................................................................................... 158
162
TOPIC: INTRODUCTION TO MEDICAL PARASITOLOGY
MEDICAL PROTOZOOLOGY
Parasitology is the study of parasites and as such does not include bacterial,
fungal or viral parasites. Human parasites are separated into intestinal and blood
borne parasites. For a parasite to be defined as intestinal it must have an intestinal
life cycle stage, though it may have life-cycle stages in the heart, circulation, lung,
tissue, other animals or the environment.
The association between two organisms may be one of the following:
MUTUALISM: mutual benefit is derived from the association.
SYMBIOSIS: mutual benefit, but the two organisms cannot live independently.
COMMENSALISM: one partner benefits (commensal) while the other (host)
is unaffected. It may be called a non-pathogenic parasite.
When an animal lives on another organism from which it receives food and shelter
without any compensation to it, and then this association is called parasitism, and
the animal, which derives advantages, is the parasite. All animals have parasites;
hence there are more parasites than free-living animals. The habitat occupied by a
parasite is very different from the environment of its free-living ancestors, hence it
has either to adapt itself to this new habitat or perish.
PARASITISM: one organism (parasite) lives at the expense of the other (host).
The latter usually suffers from the association pathogenic parasite). Parasitism is
the form of mutual relation between organisms of various kinds, from which
one (parasite) uses other (host) as environment for living and from which it
obtains food bringing him damage (disease).
PARASITOLOGY is a complex biological science studying the phenomena of
parasitism. There are mutual relation between the parasite and host, their dependence
on the factors of external environment, and also diseases, caused by the parasites,
and methods of struggle with them at the man, animals and plants.
MEDICAL PARASITOLOGY consists of 3 parts: medical protozoology,
medical helminthology and medical entomology.
CLASSIFICATION OF PARASITES
Each parasite belongs to a phylum, class, order, family, genus and species; the
scientific designation of a parasite is binomial, a generic name (genus) and a specific
name (species).
The parasites of humans in the kingdom protozoa are now classified under 3
phyla: Sarcomastigophora (containing the amoebas and flagellates); Apicomplexa
(containing the sporozoans); and Ciliophora (containing the ciliates). The important
human parasites are found within these great assemblages.
1. Class LOBOZEA (Sarcodina) is typically amoeboid and in represented in
humans by class of Entamoeba, Endolimax, lodamoeba, Naegleria, and
Acanthamoeba.
3
 2. Class ZOOMASTIGOPHORA, the flagellates, have one or more whip-like
flagella and, in some cases, an undulating membrane (e.g., trypanosomes). These
include intestinal and genitourinary flagellates (Giardia, Trichomonas,
Dientamoeba, Chilomastix) and blood arm tissue flagellates (Trypanosoma,
Leishmania).
3. Class SPOROZOA undergoes a complex life cycle with alternating sexual
and asexual reproductive phases, usually involving two different hosts (e.g., arthropod
and vertebrate, as in the blood forms). The subclass Coccidia contains the human
parasites Isospora, Toxoplasma, and others. One of these, Cryptosporidium, has
been implicated as a cause of intractable diarrhea among the immunosuppressed.
Among the Haemosporina (blood sporozoans) are the malaria parasite (Plasmodium
species) and the subclass Piroplasmia, which includes Babesia species. Pneumocystis
has recently been shown to be a member of the Fungi rather than the Protozoa. It is
another opportunistic parasite of immunosuppressed individuals.
4. Class LITOSTOMATEA is a complex protozoan bearing cilia distributed in
rows or patches, with two kinds of nuclei in each individual. Balantidium coli, a
giant intestinal ciliate of humans and pigs, is the only human parasite representative
of this group.
THE PARASITIC WORMS, or helminthes, of human bein, belong to two phyla:
1. PLATYHELMINTHES (flatworms) lack a true body cavity (celom) and are
characteristically flat in dorsoventral section. A medically important species belong
to the classes Cestoda (tapeworms) and Trematoda (flukes). The tapeworms of
humans are band-like and segmented; the flukes are typically leaf-shaped, and the
schistosomes narrow and elongate. The important tissue and intestinal cestodes
humans belong to the genera Diphyllobothrium, Spirometra, Taenia, Echinococcus,
Hymenolepis, and Dipylidium. Medically important trematode genera include
Schistosoma, Paragonimus, Clonorchis, Opistorchis, Heterophyes, Metagonimus,
Fusciolopsis, and Fasciola.
2. NEMATHELMINTHES (worm-like, separate-sexed, insegmented
roundworms) include many parasitic species that infect humans.
PHYLUM ARTHROPODA includes 3 classes of medical importance:
1. CLASS CRUSTACEA: Cyclops, etc.
2. CLASS ARACHNIDA (Octapoda): scorpions, ticks and mites.
3. CLASS INSECTA (Hexapoda): mosquitoes, flies, bugs, lice and fleas.
TYPES OF PARASITES
ENDOPARASITE: lives within the body of the host (infection).
ECTOPARASITE: lives on the outside of the body of the host (infestation).
OBLIGATE PARASITE: completely dependent upon its host and cannot lead
a free life.
4 161
Liver Flukes: ..................................................................................................................... 60
Clonorchis sinensis, Opisthorchis species, and Dicrocoelium dendriticum ................. 60
Clonorchis sinensis ............................................................................................................ 62
Metagonimus yokogawai .................................................................................................. 64
Paragonimus westermanni ............................................................................................... 66
The Schistosomes .............................................................................................................. 68
Schistosoma mansoni ........................................................................................................ 69
Schistosoma japonicum .................................................................................................... 71
Schistosoma haematobium ............................................................................................... 73
TOPIC: PLATHELMINTHES, CESTOIDEA .............................................................. 75
Larval Cestodes, which Infect Man ................................................................................ 75
Echinococcus granulosus .................................................................................................. 75
Echinococcus multilocularis ............................................................................................. 78
The Cestodes ..................................................................................................................... 79
Taenia species .................................................................................................................... 80
Hymenolepis nana ............................................................................................................. 85
Diphyllobothrium latum ................................................................................................... 86
TOPIC: NEMATHELMINTHES, NEMATODA ......................................................... 88
The Nematodes .................................................................................................................. 88
Ascaris lumbricoides ......................................................................................................... 88
Hookworm species ............................................................................................................ 90
Trichuris trichiura ............................................................................................................ 93
Strongyloides stercoralis .................................................................................................. 96
Enterobius vermicularis ................................................................................................... 99
Trichinella spiralis .......................................................................................................... 101
Wuchereria bancrofti ..................................................................................................... 104
Brugia malayi .................................................................................................................. 106
Brugia Timori .................................................................................................................. 107
Loa loa ............................................................................................................................. 108
Mansonella species .......................................................................................................... 110
Microfilaria worms found in tissue and skin ................................................................. 111
Onchocerca volvulus ........................................................................................................ 111
Mansonella streptocerca ................................................................................................. 114
Dracunculus medinensis ................................................................................................. 114
TOPIC: ARTHROPODA, ARACHNOIDEA, ACARINA ......................................... 117
Phylum Arthropoda ........................................................................................................ 117
Crustacea ......................................................................................................................... 118
The Arachnida ................................................................................................................. 119
Scorpions ......................................................................................................................... 119
Solpugids ......................................................................................................................... 120
Spiders ............................................................................................................................. 121
Mites and ticks ................................................................................................................ 124
Sarcoptes scabiei ............................................................................................................. 125
The demodex mite ........................................................................................................... 127
Ixodid ............................................................................................................................... 128
Ticks as disease vectors .................................................................................................. 129
 CONTENT FACULTATIVE PARASITE: capable of leading both a free-living and a
parasitic existence.
TOPIC: INTRODUCTION TO MEDICAL PARASITOLOGY ................................... 3 INCIDENTAL PARASITE: can establish itself in a host in which it does not
TOPIC: PROTOZOA ........................................................................................................ 6 ordinarily live.
The Amoebae ..................................................................................................................... 10 ACCIDENTAL PARASITE: a free-living organism that may live as a parasite
Entamoeba histolytica ...................................................................................................... 11 in a host.
Entamoeba coli .................................................................................................................. 13 PERMANENT PARASITE: remains all or most of its life in or on its host.
Entamoeba hartmanni ...................................................................................................... 14 TEMPORARY PARASITE (occasional or intermittent): free-living but seeks
Endolimax nana ................................................................................................................ 14
its host from time to time for food.
TOPIC: ZOOMASTIGOPHOREA ................................................................................ 15
PERIODIC PARASITE (transitory): passes a definite part of its life cycle as a parasite.
Giardia lamblia
(giardiasis) ................................................................................................................... 15
SPECIFIC PARASITE: occurs in one particular host.
Trichomonas vaginalis PSEUDOPARASITE: An artifact mistaken for a parasite.
(trichomoniasis, «trich» or «trick») .......................................................................... 18 COPROZOIC or SPURIOUS PARASITE: a free-living or a non-human
Trichomonas vaginalis, Pentatrichomonas hominis, parasite passing through the alimentary canal without infecting man or contaminating
Enteromonas hominis, his stools after being passed.
and Retortamonas intestinalis ................................................................................... 18 OPPORTUNISTIC PARASITE: occurs in patients with impaired host defense
Trichomonas hominis ........................................................................................................ 22 (immunodeficient or immunocompromised host). Such parasite does not ordinarily
Trypanosoma species ........................................................................................................ 23 produce disease in healthy immunocompetent) individuals.
Salivarian trypanosomes .................................................................................................. 23
Stercorarian trypanosomes .............................................................................................. 23 TYPES OF HOSTS
African Trypanosomiasis .................................................................................................. 23 1. FINAL HOST or DEFINITIVE HOST: harbours the adult or sexually ma-
South American trypanosomiasis .................................................................................... 27 ture parasite.
Leishmania species ............................................................................................................ 30 2. RESERVOIR HOST: An animal that harbours the same species of parasites
Visceral leishmaniasis ....................................................................................................... 30 as man and constitute a source of infection to him. Usually these animals are not
Cutaneous leishmaniasis ................................................................................................... 31
affected by infection. The reservoir host serves as a potential source of human
Mucocutaneous leishmania ............................................................................................. 33
reinfection and as a means of sustaining the parasite when it is not infecting man.
TOPIC: APICOMPLEXA, SPOROZOAE, CILIOPHORA ........................................ 36
Toxoplasma gondii ............................................................................................................ 36 Diseases of animals that are transmissible to man are called zoonotic diseases.
Malaria .............................................................................................................................. 39 3. INTERMEDIATE HOST: harbours the immature or asexual stages of the
Blood Parasites .................................................................................................................. 39 parasite.
Malaria .............................................................................................................................. 39 4. VECTOR: An arthropod that carries a parasite to its host.
Species Specific Characteristics ....................................................................................... 42
Plasmodium falciparum .................................................................................................... 42 Source and mode of parasitic infection or infestation
Plasmodium vivax ............................................................................................................. 44 1. Food and drink:
Plasmodium ovale ............................................................................................................. 46 Ingestion of raw or undercooked food or drinking water or other beverages
Plasmodium malariae ....................................................................................................... 47 containing the infective stage of the parasite.
Diagnosis of malaria parasites ......................................................................................... 47 2. Soil, dust and water (canals and streams)
Estimation of Percentage Parasitaemia of Plasmodium falciparum ............................ 50 a) Ingestion of food or drink contaminated with soil or dust containing the infective
The Ciliates ........................................................................................................................ 52 stage of the parasite.
Balantidium coli ................................................................................................................ 52 b) Inhalation of dust.
TOPIC: PLATHELMINTHES, TREMATODA ........................................................... 54
c) Direct contact with soil (handling, walking barefooted); the infective stage
Helminth Parasites ............................................................................................................ 54
penetrates the skin.
The Trematodes ................................................................................................................. 54
Intestinal and Liver Flukes: Gastrodiscoides hominis, Nanophyetus salmincola,
d) Using water streams (washing, swimming, wading, irrigation, etc.); the infective
Fasciolopsis buski, and Fasciola hepatica ................................................................. 55 stage penetrates the skin or mucous membrane.
160 5
 3. Vector To maintain a velocity of 800 miles per hour, the 0.3-gram fly would have had to
a) Bite of vector inoculating the infective stage. consume more than 150% of its body weight in food every second;
b) Feces of vector containing the infective stage contaminate the skin (intact or wounded). The fly would have produced an audible sonic boom;
c) Ingestion of vectors containing the infective stage. The supersonic fly would have been invisible to the naked eye; and
d) Direct penetration of an arthropod into the skin. The impact trauma of such a fly colliding with a human body would resemble
4. Direct contact: that of a gunshot wound.
a) Skin contact. Using the original report as a basis, Langmuir estimated the deer botfly’s true
b) Sexual contact. speed at 25 miles per hour.
c) Autoinfection or direct infection.
5. Congenital infection.

Ettect of the parasite on the host (Pathogenicity)

The effect depends on the number, size and shape of the parasite; its activity
(movement and migration); site (habitat); specific toxin and host reaction.

The effect may be due to:

1. The parasite abstracting nourishment from the host.
2. Mechanical effect leading to tissue destruction as a result of trauma, pressure,
compression or obstruction; feeding on tissues or irritation of tissues leading to
inflammatory or neoplastic reactions.
3. Toxic effects from toxins secreted or waste products excreted by the parasite,
leading to poisoning or allergic reactions.
4. Secondary infection with other organisms as bacteria. The host reaction to the
invading parasite may be:
a) Generalized in the form of fever, anaemia, leucocytosis, leucopaenia,
eosinophilia, allergic reactions, weakness, etc.
b) Localized according to the tissue or organ affected, e.g. gastrointestinal
disturbances (colic, dyspepsia, diarrhea, and dysentery), itching, ulceration,
hepatomegaly, splenomegaly, etc.

TOPIC: PROTOZOA
MEDICAL PROTOZOOLOGY
Protozoa are single celled organisms. There are four classes of Protozoa commonly
found in concentrated faecal samples. These are differentiated by the method of
motility. Protozoa include Entamoeba, Giardia, Trichomonas, Cryptosporidium,
Isospora, Pneumocystis and Balantidium. There are two diagnostic life-cycle stages
commonly seen in parasites – the cyst and the adult trophozoite stage. The trophozoite
stage is analyzed directly on a slide without concentration. Cysts require concentration.
The key diagnostic factor is that Protozoan cysts are typically 5-30 microns in
diameter, and as such are smaller than most Helminthes eggs. Due to the size they
are particularly difficult to see under the microscope if the sample clarity is bad.
6 159
particularly in the scientific and development communities. Because most people
outside of Africa learn the word later in life through formal means, e.g. books and
documentaries, the use of tsetse alone is likely to become dominant. The
pronunuciation of the word differs in different regions. Many African languages
have an ejective ts sound and so a common pronunciation of the word involves
two identical syllables both having this ts sound and a shorter sound of the vowel,
as ts-eh-ts-eh. The British pronunciation of the word uses two different sounds for
the two different syllables, generally tee-tsee. In Zimbabwe, it is generally
pronounced tseh-tsee.
Oestridae
Oestridae (also called botfly or bot fly) is a family of Oestroidea. It is one of
several families of hairy flies whose larvae live as parasites within the bodies of
mammals, such as the Desert Woodrat.
Life cycle:
! Adult botflies deposit eggs on a host body: common houseflies for example.
! Eggs are deposited on animal skin by their host: the body heat of the animal
induces hatching upon contact. Some forms of botfly also reside in the digestive
tract when consumed by a licking action.
! Myiasis: larvae burrow into the skin (or tissue lining) of the new host animal.
! Mature larvae drop from the host and complete the pupae stage in soil.
Remedies:
! Immediate contact with larvae can be remedied with alcohol.
! If one is afflicted with bot flies, one «cure» (which is really more of a folk
remedy) is to put meat over the affected area while the flies are in their larval stage,
thereby cutting off the parasites’ air supply. The grubs should then burrow through
the meat to gain access to oxygen, at which point the meat may be removed with the
larvae trapped inside.
The botfly maggot cannot be removed easily whilst alive due to the spines that
run along its body. One medical treatment is to suffocate the grub by sealing off the
air hole found in the surrounding blister. This can be done with petroleum jelly or a
similar substance. This forces the grub to expose itself, making it easier to remove.
Deer Botfly
The deer botfly was reported for many years to be the fastest of all flying insects,
cited by the New York Times and Guinness Book of World Records as traveling at
speeds of over 800 miles per hour. The source of this remarkable claim was an
article by entomologist Charles H. T. Townsend in the 1927 Journal of the New York
Entomological Society, wherein Townsend claimed to have estimated a speed of
400 yards per second while observing botflies at 12,000 feet in New Mexico. In
1938, Nobel laureate chemist Irving Langmuir examined the claim in detail and
refuted the estimate. Among his specific criticisms were:
158
The medically important Helminthes are nematodes (roundworms), cestodes
(tapeworms) and trematodes (flukes). Genera include: Fasciola, Schistosoma, As-
caris, Hookworm, Trichuris, Taenia and Enterobius. The normal stage for examina-
tion is the egg stage, although larvae may develop in some organisms (Strongy-
loides). The diameter of the eggs ranges from 30-150 microns.
The other major groupings of parasites are known as blood-borne parasites where
they are transmitted through an arthropod vector. By far the most important arthropod
for transmitting parasitic infections are the mosquitoes. Mosquitoes are known to
carry malaria and filarial nematodes. Different types of biting flies transmit African
trypanosomiasis, leishmaniasis and several kinds of filariasis.
Most protozoan and helminthic infections that are transmitted by arthropods can
readily be diagnosed, on clinical grounds alone, but are usually identified by fairly
simple techniques designed to present the presence of the causative parasite by
microscopy. Sophisticated techniques are also being employed including highly
sensitive and specific simple monoclonal antibody tests, DNA probes and PCR
primers.
Protozoa exhibit a wide variety of morphologies. There is no one shape or
morphology, which would include a majority of the protozoa. Shapes range from the
amorphous and ever-changing forms of amoeba to relatively rigid forms dictated in
part by highly ordered cytoskeletons or secreted walls or shells. Many protozoan
species exhibit complex life cycles with multiple stages. Sometimes the different
life cycle stages are so dissimilar that they have been mistaken for completely different
species.
The vast majority of protozoa are microscopic. However, they do exhibit an
incredibly large range of sizes. Extant species range in size from < 1 µm (10-6 meter)
to several mm. Fossilized Forminiferida of several cm have been identified. Most of
the organisms discussed in Parazitology will be 3-50 µm. This small size necessitates
the use of a microscope to detect protozoa. An electron microscope is needed for
detailed morphological studies.
Protozoa are found virtually everywhere. As a group, the protozoa are extremely
adaptable. Individual species, though, may have very specific-niches.
Like all other organisms, protozoa must be able to acquire and metabolize
nutrients from their environment. Many protozoa simply absorb solutes (i.e.,
osmotrophy) from their media, while some are scavengers that ingest solid material
(i.e., phagotrophy). Predatory protozoa either actively hunt down or passively ambush
other organisms (typically bacteria or other protozoa). Some protozoa are
photosynthetic and can capture the energy of the sun and convert it to usable chemical
energy (i.e., autotrophic or phototrophic). Many protozoa are not restricted to a
single feeding mechanism and can utilize combinations of the above (i.e.,
heterotrophic, mixotrophic).
Protozoa are conveniently divided into free-living and symbiotic with a few
that are facultative symbionts. Generally free-living organisms are found in the
soil or aqueous environments, whereas symbionts live in close association with another
7
organism. Symbiosis implies a physiological dependency on another organism for
its nutrition. Different forms of symbiosis can be distinguished on the nature of the
association between the dissimilar organisms:
Commensalism: denotes an interaction that is beneficial to one organism but has no
affect on the other organism. For example, many protozoa live in the alimentary canal of
another organism without harming it. These commensals are often scavengers or predators
that exploit the abundance of nutrients or bacterial fauna provided by the host organism.
Mutualism: is a special form of commensalism in which both organisms derive
some benefit and have become dependent on each other. The classic example of
mutualism is the protozoan Trichonympha found in the gut of termites. Trichonympha,
with the assistance of symbiotic bacteria, digests the wood particles (i.e., cellulose)
ingested by the termite.
Parasitism: denotes a relationship in which one organism (the parasite) benefits
at the expense of the other organism (the host). Generally this host expense implies
that the parasite takes in macromolecules from the host and releases others into
the host. In some instances the parasitism will be overtly harmful to the host and
referred to as being pathogenic. These pathogenic protozoa will be the primary
focus of this course.
The earliest observations of protozoa noted their motility. However, motility
is not a universal feature of protozoa and different protozoa utilize different
mechanisms for their movement. In fact, protozoa were initially classified based in
part on their motility. Cilia and flagella are subcellular structures, which propel
protozoa through a fluid medium. Flagella are long whip-like structures, which propel
the organism as a result of wave-like beat, which is propagated through their length.
Flagellated protozoa typically have one or a few flagella per organism. In contrast,
ciliated protozoa are usually covered with rows of numerous cilia. Cilia and flagella
can also assist in the procurement of food, reproduction and other functions.
In contrast to the swimming exhibited by flagellates and ciliates, amoeba are
protozoa that crawl along a solid substratum in a fashion known as “amoeboid
movement”. The amoeba projects out a pseudopod, or false foot, and then pull the
rest of the body forward.
The most common form of reproduction in protozoa is asexual binary fission.
In other words, a single organism will divide into two equal organisms. A slight
modification of this binary fission, called budding, is when one of the newly formed
cells is smaller than the other. Typically the larger cell is called the mother and the
smaller is the daughter. Some protozoa will form an intracellular bud and essentially
give birth. Another variation of binary fission is a multiple fission or segmentation.
In this situation, several rounds of nuclear replication occur without cytokinesis.
This multinucleated cell will then form multiple progeny simultaneously.
In summary, protozoa are unicellular eukaryotic microorganisms. However,
the amount of diversity in terms of morphology, size and life styles exhibited by
protozoa makes it difficult to develop a more precise definition. However, protozoa
do exhibit many of the features found in other eukaryotes.
8 157
mans. Trypanosoma congolense and Trypanosoma vivax are the two most important
species infecting bovine cattle in sub-saharan Africa. Trypanosoma simiae causes a
virulent disease in swine. Other forms of animal trypanosomiasis are also known
from other areas of the globe, caused by different species of trypanosomes and trans-
mitted without the intervention of tsetse.
Tsestse control: Tsetse control has been undertaken in order to reduce the
incidence of the diseases, which the flies transmit. Two alternative strategies have
been used in the attempts to reduce the African trypanosomiases. One tactic is
primarily medical or veterinary and targets the disease directly using monitoring,
prophylaxis, treatment, and surveillance to reduce the number of organisms, which
carry the disease. The second strategy is generally entomological and intends to
disrupt the cycle of transmission by reducing the number of flies. The idea of tsetse
control implies a change in the relationship between people and these insects. Prior
to the twentieth century, people in Africa had largely adapted to the presence of
tsetse. Human settlement patterns and agricultural practices had adapted to the
presence of the fly. For example, in Ethiopia draft powered framing was restricted to
the highland areas where the flies were absent whereas lowland areas where tsetse
are present were more sparsely populated by people living a nomadic, less
agriculturally intensive lifestyle. Tsetse control is a response to changing conditions.
Tsetse control has been proposed as a way of reducing the incidence of the disease
in the populations living in tsetse regions, of allowing the expansion of human
settlement and agriculture into new areas, and of helping people previously relocated
either in forced transfers or due to migration. Tsetse control efforts have been
undertaken throughout the African continent but long-term, sustainable control has
rarely been achieved. Tsetse control efforts invariably are tied to the complex problems
of poverty, heath, politics, and violence, which have proved such a disaster for the
African people. The reduction of fly numbers has generally been attempted with two
different aims, either eradication which intends to completely eliminate tsetse from
the area or control, which aims simply to reduce the numbers. Eradication is an idea
which has often been imagined, has repeatedly been attempted, and is still proposed
but many reasons suggest that control is a safer, cheaper, more realistic, and sustainable
approach. Eradication refers to the successful killing of every tsetse either in a region
or, under more grandiose proposals, from the entire African continent. Local
eradication efforts have repeatedly been undertaken and have achieved temporary
success only to fail in the long term because tsetse were able to re-invade (Sansibar).
All of the economic, ecological, political, and environmental justifications for
eradication have been called into question. The economic justification for eradication
offsets the immense costs of the eradication campaign against the medical and
veterinary benefits, which are considered to accrue in perpetuity.
Etymology: The word “tsetse” comes from Tswana, a language of southern
Africa, and, in that language, the word means fly. Because of this meaning, the
phrase “tsetse fly” is redundant. This is an example of «incomplete incorpora-
tion». Recently “tsetse” without the “fly” has become more common in English,
the lymphatic system, then into the bloodstream, and eventually into the brain. The
disease causes the swelling of the lymph glands, emaciation of the body, and eventu-
ally leads to death. Uninfected tsetse may bite the infected animal prior to its death
and acquire the disease, thereby closing the transmission cycle. The tsetse vectored
trypanosomiases affect various vertebrate species including humans, antelopes, bovine
cattle, camels, horses, sheep, goats, and pigs. These diseases are caused by several
different trypanosome species, which may also survive in wild animals such as
crocodiles and monitor lizards. The diseases have different distributions across the
african continent and are therefore transmitted by different species of tsetse.
Tsetse vectored human trypanosomiases: Human African trypanosomiasis, also
called sleeping sickness, is caused by trypanosomes of the Trypanosoma brucei
species. This disease is invariably fatal unless treated but can almost always be cured
with current medicines, if the disease is caught early enough. Sleeping sickness begins
with a tsetse bite leading to an inoculation in the sub-cutaneous tissue. The infection
moves into the lymphatic system leading to a characteristic swelling of the lymph
glands, which is called Winterbottoms’s sign. The infection progresses into the blood
stream and eventually crosses into the central nervous system and invades the brain
leading to extreme lethargy and eventually to death. The page on human sleeping
sickness has more extensive information about this disease. The Trypanosoma brucei
species, which causes the disease, has often been subdivided into three sub-genera,
which were identified based either on the vertebrate hosts, which the strain could
infect or on the virulence of the disease in humans. The trypanosomes infectious to
animals and not to humans were named Trypanosoma brucei brucei. The strains
which infected humans were divided into two sub-species based on their different
virulences: Trypanosoma brucei gambiense was thought to have a slower onset and
Trypanosoma brucei rhodesiense refers to strains with a more rapid, virulent onset.
This characterization has always been problematic but was the best that could be
done given the knowledge of the time and the tools available for identification. A
recent molecular study using restriction fragment length polymorphism analysis
suggests that the three sub-genera are polyphyletic, so the elucidation of the strains
of T. brucei infective to humans will require a more complex explanation. Other
forms of human trypanosomiasis also exist but are not transmitted by tsetse. The
most notable is American trypanosomiasis, known as Chagas disease, which occurs
in South America, caused by Trypanosoma cruzi, and transmitted by certain species
of the reduviidae, members of the true bugs.
Tsetse vectored animal trypanosomiases: Animal trypanosomiasis, also called
nagana when it occurs in bovine cattle or horses or sura when it occurs in domestic
pigs, is caused by several trypanosome species. These diseases reduce the growth
rate, milk productivity, and strength of farm animals, generally leading to the
eventually death of the infected animals. Certain species of cattle are called
trypanotolerant because they can survive and grow even when infected with
trypanosomes although they also have lower productivity rates when infected. The
course of the disease in animals is similar to the course of sleeping sickness in hu-
156
PROTOZOAN TAXONOMY
Taxonomy is concerned with the classification and naming of organisms. In
addition to nomenclature, taxonomy also functions to place organisms into groups
sharing common features and presumably evolutionary relationships. However,
taxonomic criteria are often arbitrary and taxonomy is always changing to reflect
new discoveries and interpretations.
Protozoa are usually classified within the Kingdom Protista (Haeckel, 1866),
which includes other unicellular eukaryotes.
Historically protozoa were divided into four major groups: the amoeba, the
flagellates, the sporozoa and the ciliates. The distinguishing features between the
groups were based on motility (i.e., amoeboid, flagella, cilia). The sporozoa were a
heterogeneous group that produced spores during one stage of their life cycles and
exhibited “gliding” motility.
In addition, some members within these two groups share some common features
and therefore the amoeba and flagellates can be grouped together (e.g.,
sarcomastigophora).
So, the parasites of humans in the subkingdom PROTOZOA are now classified
under 3 phyla: SARCOMASTIGOPHORA (containing the amoebas and
flagellates); APICOMPLEXA (containing the sporozoans); and CILIOPHORA
(containing the ciliates). The important human parasites are found within these great
assemblages.
1. Class LOBOZEA (Sarcodina) is typically amoeboid and is represented in
humans by species of Entamoeba, Endolimax, Iodamoeba, Naegleria, and
Acanthamoeha.
2. Class ZOOMASTIGOPHORA, the flagellates, have one or more whip-like
flagella and, in some cases, an undulating membrane (e.g., trypanosomes). These
include intestinal and genitourinary flagellates (Giardia, Trichomonas,
Dientumoeba, Chilomastix) and blood and tissue flagellates (Trypanosoma,
Leishmania).
3. Class SPOROZOA undergoes a complex life cycle with alternating sexual
and asexual reproductive phases, usually involving two different hosts (e.g., arthropod
and vertebrate, as in the blood forms). The subclass Coccidia contains the human
parasites Isospora, Toxoplasma, and others. Among the Haemosporina (blood
sporozoans) are the malaria parasites (Plasmodium species).
4. Class CILIOPHORA is a complex protozoon bearing cilia distributed in
rows or patches, with two kinds of nuclei in each individual. Balantidium coli, a
giant intestinal ciliate of humans and pigs, is the only human parasite representative
of this group.
GENERAL MORPHOLOGY OF PROTOZOA
1. Protozoa are unicellular organisms (single cell) sometimes called non-cellular
or acellular, being not divided into cells.
2. Each protozoon performs all functions of life.
9
 3. The protozoon is made of a mass of protoplasm differentiated into cytoplasm
and nucleoplasm.
4. The cytoplasm consists of outer thin hyaline ectoplasm and inner voluminous
granular endoplasm.
5. The ectoplasm functions in: protection, locomotion, and ingestion of food,
excretion and respiration.
6. The endoplasm is concerned with metabolism. It encloses:
a) food vacuoles: containing food during digestion,
b) volutin granules: stored food in the form of carbohydrate (glycogen vacuoles)
or protein (chromatoid bodies),
c) excretory vacuoles: collect waste products and discharge them to the exterior
by bursting through the ectoplasm or by an anal opening (cytopyge),
d) the nucleus.
7. The nucleus functions in reproduction and maintaining life. It is made of nuclear
membrane, nuclear sap, chromatin. In the vesicular nucleus the chromatin is
concentrated in a mass (the karyosome) or distributed between the karyosome and
the inner surface of the nuclear membrane (peripheral chromatin). In the massive
nucleus the chromatin is distributed diffusely.
GENERAL BIOLOGY OF PROTOZOA
1. Nutrition either by: absorption of liquid food through the body surface, or
ingestion of solid particles by the help of pseudopodia or through the cytostome.
2. Excretion either by: diffusion through the body surface, or excretory vacuoles.
3. Secretions: digestive enzymes, toxins, and material for cyst wall, enzyme to
liquefy tissues.
4. Respiration either: aerobic or anaerobic.
5. Locomotion either by:
a) Pseudopodia (amoeboid movement).
b) Flagella: whip-like filaments arise from the kinetoplast+ blepharoplast +
parabasal body).
c) Cilia: like flagella but smaller and more numerous covering most of the body.
6. Cyst formation: encystment of some protozoa is essential for survival outside
the body of the host and during transmission from host to host.
7. Reproduction may be asexual or sexual.
a) Asexual reproduction (simple fission): division of the nucleus by amitosis (simple
division) or mitosis (differentiation of the chromatin into chromosomes); division of the
cytoplasm by simple fission into two (binary fission) or more (multiple fission).
b) Sexual reproduction: the formation of 2 cells (male and female gametes) by
reduction division, and their union (or syngamy) resulting in the formation of a zygote.
The Amoebae
Amoebae are characterized by possessing clear protoplasm, which form
pseudopodia. These pseudopodia are the means by which these organisms move and
10
behavior or have other effects which improve the chances of transmission and sur-
vival. These trypanosomes have become highly evolved and developed a life cycle,
which requires periods in both the vertebrate and tsetse hosts. Tsetse transmit
trypanosomes in two ways, mechanical and biological transmission.
! Mechanical transmission involves the direct transmission of the same individual
trypanosomes taken from an infected host into an uninfected host. The name
mechanical reflects the similarly of this mode of transmission to the transmission
which could be caused mechanically with a syringe. Mechanical transmission requires
that tsetse feed on an infected host and acquire trypanosomes in the bloodmeal, and
then, within in a relatively short period, for tsetse to feed on an uninfected host and
regurgitate some of the infected blood from the first bloodmeal into the tissue of the
uninfected animal. This type of transmission occurs most frequently when tsetse are
interrupted during a bloodmeal and attempt to satiate themselves with another meal.
Other flies, such as horse-flies, also can cause mechanical transmission of
trypanosomes.
! Biological transmission requires a period of incubation of the trypanosomes
within the tsetse host. The term biological is used because trypanosomes must
reproduce through several generations inside the tsetse host during the period of
incubation, which requires extreme adaptation of the trypanosomes to their tsetse
host. In this mode of transmission, trypanosomes reproduce through several
generations, changing in morphology at certain periods. This mode of transmission
also includes the sexual phase of the trypanosomes. Tsetse are believed to be more
likely to become infected by trypanosomes during their first few bloodmeals. Tsetse
infected by trypanosomes are thought to remain infected for the remainder of their
lives. Because of the adaptations required for biological transmission, trypanosomes,
which are transmitted biologically by tsetse cannot be transmitted in this manner by
other insects.
The relative importance of these two modes of transmission for the propagation
of tsetse-vectored trypanosomiases is not yet well understood. However, since the
sexual phase of the trypanosome lifecycle occurs within the tsetse host, biological
transmission is a required step in the life cycle of the tsetse vectored trypanosomes.
The cycle of biological transmission of trypanosomiasis involves two phases, one
inside the tsetse host and the other inside the vertebrate host. Trypanosomes are not
passed between a pregnant tsetse and her offspring so all newly emerged tsetse adults
are free of infection. An uninfected fly, which feeds upon an infected vertebrate
animal may acquire trypanosomes in its proboscis or gut. These trypanosomes,
depending on the species, may remain in place, move to a different part of the digestive
tract, or migrate through the tsetse body into the salivary glands. When an infected
tsetse bites a susceptible host, the fly may regurgitate part of a previous bloodmeal,
which contains trypanosomes or may inject trypanosomes contained within its saliva.
It is believed that the inoculation must contain a minimum of 300 to 450 individual
trypanosomes to be successful, and may contain up to 40,000 individuals. The
trypanosomes are injected into vertebrate muscle tissue but make their way, first into
155
 Tsetse metabolism: Tsetse metabolism consists of ingesting vertebrate blood,
which is called hematophagy, and digesting this blood to obtain energy and biomass.
Tsetse have specialized cells that contain bacterial endosymbionts required for
survival. An unusual aspect of tsetse metabolism is the particular pathway which
tsetse use for flight, which seems to be responsible for the extremely high energy
output and elevated flying speeds which tsetse can achieve. Most insects, such as
honey bees, consume sugar predominantly for metabolic energy. Tsetse, instead, use
a pathway which involves the conversion of the amino acids proline and alanine.
The result of this pathway is that tsetse can create large amounts of ATP but can only
sustain this metabolic output for short durations. Tsetse therefore fly at very high
speeds (they are known to be able to follow a car moving at thirty miles an hour) but
can only sustain their flight for short durations of around thirty seconds.
General biology: Remarkably the Tsetse Glossina palpalis is also a vector and
host of Hepatozoon petti, a parasitic Sporozoa of the nile crocodile.
Tsetse systematics: Tsetse include up to thirty-four species and sub-species
depending on the particular classification used. Tsetse are sufficiently different in
appearance and behavior to have been placed in their own distinct branch of the
flies. This placement is controversial. The science of systematics is currently struggling
to reconcile the traditional form of biological classification with the modern
understanding of genomic evolution and speciation. The controversy surrounding
the placement of tsetse is therefore likely to continue into the future. All current
classifications place all the tsetse species in a single genus named Glossina. Most
classifications place this genus as the sole member of the family Glossinidae. The
Glossinidae are generally placed within the infraorder Cyclorrhapha, which includes
the housefly and the blowfly due to the similarity of their developmental biology.
This infraorder in turn, is part of the sub-order Brachycera, the stubby flies with
reduced antenna. Tsetse are evidently part of the order Diptera, the flies.
Tsetse as vectors of trypanosomiases: Tsetse are biological vectors of
trypanosomes meaning that tsetse, in the process of feeding, acquire and then transmit
small, single-celled organisms called trypanosomes from infected vertebrate hosts
to uninfected animals. Some tsetse transmitted trypanosome species cause
trypanosomiasis, an infectious disease. In humans, tsetse transmitted trypanosomiasis
is called sleeping sickness. In animals, tsetse vectored trypanosomiases include
nagana, souma, and surra according to the animal infected and the trypanosome
species involved, although the usage is not strict and nagana is occasionally used
for any form of animal trypanosomiasis. Trypanosomes are animal parasites,
specifically protozoa of the genus Trypanosoma. These organisms are approximately
the size of red blood cells. Different species of trypanosomes infect different hosts
as can be seen in the table attached to this section. Trypanosomes range widely in
their effects on the vertebrate hosts. Some species, such as Trypanosoma theileri, do
not seem to cause any health problems except perhaps in animals, which are already
quite sick. Some strains are much more virulent. Tsetse seem to be unaffected by the
infection of trypanosomes but it is entirely possible that the parasites alter tsetse
154
engulf bacteria and red blood cells for feeding purposes. The most common amoe-
bae seen in the intestinal tract are Entamoeba histolytica, Entamoeba coli, Enta-
moeba hartmanni, Endolimax nana and Iodamoeba.
All but Entamoeba histolytica are thought to be non-pathogenic. The cysts can
be identified in an ethyl acetate concentrate by the addition of iodine to reveal the
characteristic inclusions and also by measuring the cyst using an eyepiece graticule.
The trophozoites can be seen in a fresh saline preparation of the stool although
accurate identification is on a permanently stained faecal smear.
Entamoeba histolytica
Introduction
There are a large number of species of amoebae which parasitise the human
intestinal tract. Of these Entamoeba histolytica is the only species found to be
associated with intestinal disease (Fig. 1). Although many people harbour this
organism world wide, only about 10% develop clinically invasive disease thus the
parasite has been shown to present as two very differing clinical presentations.
1. The commensal or non-invasive luminal form where the parasite causes no
signs or symptoms of disease.
2. The pathogenic or invasive form where the parasite invades the intestinal
mucosa and produces dysentery or amoebomas and may give rise to extra-intestinal
lesions via the blood, mainly to the liver.
Sargeaunt and Williams (1978) conclusively proved that invasive and non-
invasive strains of E. histolytica could be differentiated by isoenzyme electrophoresis
and the application of molecular biology has confirmed the presence of two distinct
species with the same morphological features. The pathogenic or invasive species
has retained the name E. histolytica and the non-pathogenic, non-invasive species
has been named E. dispar.
Morphology of Trophozoites
The trophozoites of E. histolytica / dispar recovered from dysenteric stools exhibit
ingested red blood cells and clear pseudopodia. Those of E. dispar will have no ingested
red blood cells. They can be up to 60m in diameter and motility is rapid and unidirectional.
On a permanently stained faecal smear e.g. Trichrome or Iron haematoxylin, the
morphological features are more visible. When using Trichrome stain nuclei, chromidial
bars, chromatin, red cells and bacteria stain red cytoplasm stains blue-green and
background and yeasts stain green. The presence of a small centrally placed karyosome
is clearly visible. With Iron haematoxylin, nuclear chromatin and the karyosome will be
stained immensely black. The remainder will be varying shades of grey/black.
Morphology of cysts
Cysts of E. histolytica / dispar are 10-15m in diameter and contain 1-4 nuclei.
Chromatoid bodies are usually present in young cysts as elongated bars with bluntly
rounded ends. Glycogen is usually diffuse, but in young cysts it is often present as a
concentrated mass, staining reddish brown with iodine.
11

Figure 1. Entamoeba histolytica
Clinical Disease
Amoebiasis is an infection usually caused by the pathogenic Entamoeba histolytica /
dispar, and is commonly an infection of the colon. It has a world wide distribution where
environmental sanitation is poor. The parasite may behave as a commensal (causing no harm
12
recognizable shape or bauplan so they can usually be distinguished without trouble
from other flies. Tsetse have large heads, distinctly separated eyes, and unusual an-
tennae. The tsetse thorax is quite large, while the abdomen is wide rather than elon-
gated and shorter than the wings. Four characteristics definitively separate adult
tsetse from other kinds of flies:
Proboscis – Tsetse have a distinct proboscis, a long thin structure attached to the
bottom of the head and pointing forward.
Folded wings – When at rest, tsetse fold their wings completely one on top of
the other.
Hatchet cell – The discal medial («middle») cell of the wing has a characteristic
hatchet shape resembling a meat cleaver or a hatchet.
Branched arista hairs – The antennae have arista with hairs, which are
themselves branched.
Tsetse anatomy: Like all other insects tsetse flys have an adult body is comprised
of three, visibly distinct, parts: the head, the thorax, and the abdomen. The head has
large eyes, distinctly separated on each side, and a distinct, forward-pointing proboscis
attached underneath by a large blub. The thorax is large, made of three fused segments.
Three pairs of legs are attached to the thorax, as are two wings and two halteres. The
abdomen is short but wide and changes dramatically in volume during feeding. The
internal anatomy of tsetse is fairly typical of the insects. The crop is large enough to
accommodate a huge increase in size during the bloodmeal since tsetse can take a
bloodmeal weighing as much as themselves. The reproductive tract of adult females
includes a uterus, which can become large enough to hold the thrid instar larva at the
end of each pregnancy.
The tsetse life cycle: Tsetse have an unusual life cycle which may be due to the
richness of their food source. Female tsetse only fertilize one egg at a time and retain
each egg within their uterus to have the offspring develop internally during the first
larval stages, a strategy called adenotrophic viviparity. During this time, the female
feeds the developing offspring with a milky substancem, which is secreted by a
modified gland in the uterus. In the third larval stage, the tsetse larva finally leave
the uterus and begin their independent life. However, the newly independent tsetse
larva simply crawls into the ground, forms a hard outer shell called the puparial case
in which it completes its morphological transformation into an adult fly. This lifestage
has a variable duration, generally twenty to thirty days, and the larva must rely on
stored resources during this time. The importance of the richness of blood to this
development can be seen since all tsetse development prior to the emergence from
the puparial case as a full adult occurs without feeding based only on nutritional
resources provided by the female parent. The female must obtain enough energy for
her needs, for the needs of her developing offspring, and to store the resources,
which her offspring will require until it emerges as an adult. Technically these insects
undergo the standard development process of insects, which comprises oocyte for-
mation, ovulation and fertilization, development of the egg, five larval stages, a
pupal stage, and the emergence and maturation of the adult.
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 Tsetse
Tsetse are large biting flies from Africa, which live by feeding on the blood of
vertebrate animals. Tsetse include all the species in the genus Glossina, which are
generally placed in their own family Glossinidae and which belong to the order
Diptera. The genus name is attributed to Wiedemann, who named the type species
Glossina longipalpis in 1830. Tsetse have been extensively studied because they are
biological vectors of the African trypanosomiases, deadly diseases, which include
sleeping sickness in humans and nagana in cattle. Tsetse are crudely similar to other
large flies, such as the housefly, Musca domestica, but can be distinguished by four
characteristics of their anatomy, two of which are easy to observe. Tsetse fold their
wings completely when they are resting so that one wing rests directly on top of the
other over their abdomen. Tsetse also have a long proboscis which extends directly
forward and is attached by a distinct bulb to the bottom of their head. Tsetse have
existed in the modern morphological form for at least 34 million years since fossil
tsetse have been recovered from the Florissant Fossil Beds in Colorado.
Tsetse biology: The biology of tsetse is relatively well understood (Fig. 45).
Tsetse have been extensively studied because of their medical, veterinary, and
economic importance, because the flies can be raised in a laboratory, and because
the flies are relatively large facilitating their analysis. Entomologists have discovered
a great deal about tsetse morphology, anatomy, development and metabolism.
Tsetse morphology: Tsetse can be seen as independent individuals in three forms:
as third instar larva, as puparia, and as adults. Tsetse first become separate from
their mothers during the third larval instar, during which they have the typical
appearance of maggots. However, this life stage is short, lasting at most a few hours,
and is almost never observed outside of the laboratory. Tsetse next become puparia—
small, hard shelled, oblongs with two distinctive, small, dark lobes at one end. Tsetse
puparia are under 1.0 centimeters long. Tsetse then emerge as adult flies. Tsetse
adults are relative large flies, with lengths of 0.5 to 1.5 centimeters, and have a
Figure 45. Tsetse fly
152
to the host) or it may act as a parasite (harming the host). It is a disease of human beings,
although some monkeys can become infected and the infection is then transmissable to humans.
Intestinal disease
Patients with intestinal disease may exhibit a number of symptoms including profuse
diarrhoea with blood and mucus, fever and dehydration. Amoebic ulcers may develop in
the large colon and can also be found in the rectal area. The ulcers are usually «flask
shaped» with a small opening on the mucosal surface and a larger area below the surface.
Hepatic Disease
Trophozoites are transported from the intestine to the liver and liver disease is
characterised with abdominal pain, fever, hepatomegaly and tenderness. If the abscess
ruptures, there is spreading to the brain, pericardium and other sites. If left untouched
the abscess will grow normally until it reaches a surface where it can discharge, e.g.
the skin, the peritoneum, the pleural cavity or the pericardium. The stretching of the
liver is presumably the main source of the pain.
Microscopy
Where amoebic dysentery is suspected, the laboratory should be informed that a
«hot stool» is being supplied so that it can be examined within twenty minutes of
being passed. On cooling the amoebae stop moving which then become very difficult
to identify. Direct microscopy should be done by mixing a small amount of the
specimen in 0.9% sodium chloride solution. This permits detection of motile
trophozoites of Entamoeba histolytica / dispar and can also provide information on
the content of the stool i.e. the presence of leucocytes and red blood cells. One
search e.g. primarily for cysts, not for amoebae, several stool samples are required
to be examined, by direct microscopy and a sensitive concentration technique. Three
negative stool samples are required before it can be accepted that there is no amoebic
infection. Microscopic examination of an amoebic abscess aspirate e.g. in the liver
or lungs, may reveal haematophagous trophozoites. It must be examined immediately
by mixing a drop of warm saline with some aspirated pus on a microscope slide.
Serology
If visceral or hepatic amoebiasis is suspected serological tests should be done, as
microscopic methods do not always reveal the characteristic trophozoites. The tests
of choice are indirect fluorescent antibody test (IFAT), counter immunoelectrophoresis
(CIEP) and enzyme linked immunosorbent assay (ELISA)
The search for E. histolytica / dispar is mainly carried out in Europe and North
America, as there is a natural concern to ensure that patients, even in the absence of
symptoms are not harbouring parasites that may lead to serious complications later on.
Entamoeba coli
Introduction
Entamoeba coli is a non-pathogenic amoeba with world wide distribution. Its
life cycle is similar to that of E. histolytica but it does not have an invasive stage and
does not ingest red blood cells.
13
 Morphology of Trophozoite
The trophozoite is larger than that of E. histolytica ranging from 15-50m in
diameter. It exhibits blunt pseudopodia with sluggish movement. A permanently
stained preparation shows a nucleus with a moderately large eccentric karyosome
with the chromatin clumped on the nuclear membrane. The cytoplasm appears
granular containing vacuoles with ingested bacteria and other food particles.
Morphology of Cysts
Cysts of E. coli are 15-30m in diameter and contain 1-8 nuclei with irregular
peripheral chromatin: karyosomes not central. Chromatoid bodies are not frequently
seen but when present they are usually splinter-like with pointed ends. Glycogen is
usually diffuse but in young cysts is occasionally found as a well-defined mass,
which stains reddish brown with iodine.
Laboratory Diagnosis
Laboratory diagnosis is made by finding the characteristic cysts in iodine stained,
formol-ether concentration method or by detecting the characteristic trophozoites in
a wet preparation or a permanent stained preparation.
Entamoeba hartmanni
Introduction
Entamoeba hartmanni is a non-pathogenic amoeba with world wide distribution.
Its life cycle is similar to that of E. histolytica but it does not have an invasive stage
and does not ingest red blood cells.
Morphology of trophozoites
Morphology of the trophozoites is similar to those of E. histolytica / dispar but
they do not contain ingested red blood cells and the motility is less rapid.
Morphology of cysts
Cysts of E. hartmanni 7-9m m in diameter and contain 1-4 nuclei. Chromatoid
bodies are usually present in young cysts as elongated bars with bluntly rounded
ends. Glycogen is usually diffuse, but in young cysts it is often present as a
concentrated mass, staining reddish brown with iodine.
Laboratory Diagnosis
Laboratory diagnosis is made by finding the characteristic cysts in iodine stained,
formol-ether concentration method or by detecting the characteristic trophozoites in
a wet preparation or a permanent stained preparation.
Endolimax nana
Introduction
Endolimax nana is a small non-pathogenic amoeba with world wide distribution.
Its life cycle is similar to that of E. histolytica but is non-invasive.
Morphology of trophozoite
Trophozoites of E. nana measure from 6-12mm. Motility is sluggish with blunt
14
the female is receptive for mating. The male mounts her from the back to inject
sperm. Normally the female mates only once, storing the sperm to use it repeatedly
for several sets of eggs. Males are territorial: they defend a certain territory against
other males and try to mount any females that enter that territory. The flies depend
on warm temperatures; generally, the warmer the temperature the faster the flies will
develop. In the winter, most of them survive in the larval or pupa stage in some
protected warm location. Some species of wasps can parasitize and kill the pupae.
Typical behaviors: Houseflies can only take in liquid foods. They spit out sa-
liva on solid foods to pre-digest it, and then suck it back in. They also throw up
partially digested matter and eat it again. The flies can walk on vertical planes, and
can even hang upside down from ceilings. This is accomplished with the surface
tension of liquids secreted by glands near their feet. Lacking eyelids, the flies con-
tinually clean their eyes with their forelegs. Most of their taste and smell sensor cells
are on hairs on their legs, this is why they also keep rubbing their legs together. Flies
have a very highly-evolved evasion reaction which helps to ensure their survival. It
is possible to confuse a fly´s evasion system by swatting it with two objects simulta-
neously from different directions. The holes in a fly swatter minimise the air current
which warns the fly of being hit, whilst reducing air resistance and increasing speed
of the swat.
Sex determination mechanism: The housefly is an object of biological research,
mainly because of one remarkable quality: the sex determination mechanism.
Although a wide variety of sex determination mechanisms exists in nature (e.g. male
and female heterogamy, haplodiploidy, environmental factors) the way sex is
determined is usually fixed within one species. However, the housefly exhibits many
different mechanisms for sex determination, such as male heterogamy (like most
insects and mammals), female heterogamy (like birds) and maternal control over
offspring sex. This makes the housefly one of the most suitable species to study the
evolution of sex determination.
Evolution: Even though the order of flies (Diptera) is much older, true houeseflies
evolved in the beginning of the Cenozoic era, some 65 million years ago. They
probably originated in what is today Africa and spread to Europe and Asia. They are
not native to the Americas and it is quite possible that they were introduced by
Christopher Columbus or other early seamen.
Flies and humans: In colder climates, houseflies only occur together with humans.
They have a tendency to aggregate and are difficult to dispel. They are capable of
carrying over 100 pathogens, such as typhoid, cholera, Salmonella, bacillary
dysentery, tuberculosis, anthrax ophthalmia, and parasitic worms. The flies in poorer
and lower-hygiene areas usually carry more pathogens. Some strains have become
immune to common insecticides. In art, extremely life-like houseflies have sometimes
been depicted in the trompe l’oeil paintings of the 15th century. An example is the
painting Portrait of a Carthusian by Petrus Christus, showing a fly sitting on a fake
frame. In 2001, Garnet Hertz produced an art project in which a complete web server
was implanted into a dead fly.
151
earthworms, and small vertebrates. Some horse-fly species are known to transmit
disease and/or parasites. Chrysops is a biologic vector of Loa loa, transmitting this
filariasis between humans. A common problem in some animals, though, when large
flies are abundant, is blood loss. Some animals have been known to lose up to 300
ml of blood in a single day, which can severely weaken or even kill them.
Citation
Although the tsetse flies were responsible for transmitting sleeping sickness in
most areas, occasionally an epidemic occurred in which the disease might be conveyed
to cattle by direct contact with the ordinary horse fly, tanidae. This probably occurred
when swarms of these flies surrounded the wretched animals. In one such epidemic
some 3000 head of cattle died of trypanosomal disease in northern Rhodesia. Sir
David and Lady Bruce returned to England in 1913. David Bruce reported the results
achieved by this Sleeping Sickness Commission of the Royal Society in the Croonian
Lectures in 1915.
Musca domestica
The housefly (also house-fly or house fly) (Musca domestica) is the most
common fly occurring in homes and indeed one of the most widely distributed animals
and the most familiar of all flies; it is a pest that can facilitate serious diseases.
Physical description: The adults are 5-8 mm long. Their thorax is greyish, with
four dark longitudinal lines on the back. The underside of the abdomen is yellowish.
The whole body is covered with hair. They have reddish compound eyes. The females
are slightly larger than the males and have a much larger space between the eyes.
Like most Diptera, houseflies have only one pair of wings; the hind pair is reduced
to small halteres that aid in flight stability. Characteristically, the fourth long vein of
the wing shows a sharp upward bend.
Species that appear similar to the housefly include:
! The lesser house fly (Fannia canicularis), somewhat smaller and more slender
than M. domestica, fourth long vein of the wing is straight.
! The stable fly (Stomoxys calcitrans) looks similar to M. domestica but has a
longer piercing mouthpart, used to penetrate the skin of humans and animals in
order to suck blood.
Life cycle: Each female fly can lay up to 500 eggs (in five batches of 100 eggs
each). The eggs are white at about 1.2 mm in length. Within a day, the larvae (maggots)
hatch from the eggs; they live and feed in (usually dead and decaying) organic material,
such as garbage or feces. They are pale whitish, 3-9 mm long, thinner at the mouth
end, and have no legs. After several molts, the maggots crawl to a dry cool place and
transform into pupae, colored reddish or brown and about 8mm long. The adult flies
then emerge from the pupae. (This whole cycle is known as complete metamorpho-
sis.) The adults live from half a month to a month. After having emerged from the
pupae, the flies cease to grow; small flies are not young flies but the result of little
food during the maggot stage. Some 36 hours after having emerged from the pupa,
150
hyalin pseudopodia. In a permanently stained preparation, the nucleus exhibits a
large karyosome with no peripheral chromatin on the nuclear membrane.
Morphology of cysts
Cysts of E. nana are 6-9m in diameter. They can be spherical or ovoid in shape
and contain 4 pinpoint nuclei, which are highlighted by the addition of iodine.
Chromatoid bodies are not found and glycogen is diffuse.
Laboratory Diagnosis
Laboratory diagnosis is made by finding the characteristic cysts in iodine stained,
formol-ether concentration method or by detecting the characteristic trophozoites in
a wet preparation or a permanent stained preparation.
Entamoeba gingivalis
Entamoeba gingivalis is an Entamoeba histolytica-like amoebae that lives in/on the
teeth, gums, and sometimes tonsils. It measures 10-35 micrometers in length. Endocytotic
vacuoles are often numerous and the parasite will ingest bacteria, leukocytes, and
erythrocytes (dark circles in trophozoites, above) although it is not itself invasive. No
cysts are formed and transmission is entirely by oral-oral contact. Multiple samplings
reveal the parasite to colonize the oral cavity of nearly all adult humans.
TOPIC: ZOOMASTIGOPHOREA
Giardia lamblia
(giardiasis)
Introduction
Giardia lamblia is a flagellate of world wide distribution. It is more common in
warm climates than temporal climates. It is the most common flagellate of the intestinal
tract, causing Giardiasis (Fig. 2). Humans are the only important reservoir of the
infection. The infection is most common in parts of the world where sanitation is at
its lowest. Giardiasis is an infection of the upper small bowel, which may cause
diarrhoea. Only Giardia spreads disease.
Morphology of trophozoites
The trophozoites of G. lamblia are flattened pear shaped and are an average size
of 15 mm long, 9 mm wide and 3 mm thick. When stained, the trophozoite is seen to
have 2 nuclei, 2 slender median rods (axostyles), and 8 flagella arising from the
anterior end. They have been described as looking like tennis rackets without the
handle (they are often seen has having a comical face-like appearance when looking
at the front view).
The movement of the trophozoites is described as tumbling leaf motility, using
their 4 pairs of flagella for locomotion. They attach themselves to the surface of the
jejunal or duodenal mucosa by their disc-like suckers, which are found on their
ventral surface. They multiply in the gut by binary fission. Once the trophozoites
drop off the mucosal surface they are normally carried in the intestinal contents
down the gut where they usually encyst.
15
 Figure 44. Horse flies

deer-flies, genus Chrysops, also known as banded horse-flies because of their coloring.
Both these genera give their names to subfamilies. The «Blue Tail Fly» in the
eponymous song was probably a tabaninid common to the southeastern United States.
Adult horse-flies feed on nectar and other plant juices, but only the females also feed
on blood. Males lack the necessary mouth apparatus to do so. Most horse-flies feed
on mammal blood, but some species are known to feed on birds, amphibians or
reptiles. The females’ primary sense for locating prey is sight, and they have large,
compound eyes that serve this purpose well. The flies usually lay waiting in shady
areas for prey to happen by. They are attracted to large, dark objects, and to certain
animal odors and carbon dioxide. They are also attracted by motion, their eyes being
well adapted to its detection. The eyes of horse-flies are generally brightly colored,
and this coloration is the primary means entomologists use to sex them. A horse-fly’s
bite can be very painful. Unlike insects that pierce the skin, horse-flies have
mouthparts that work like miniature knives, which they use to slash open the skin
with a scissor-like motion. This causes the blood to seep out as the horse-fly licks it
up. While some horse-flies are known to have venom, none is known to be dangerous
to humans. When attacking humans, the flies generally prefer the head and upper
body regions, going unnoticed until a bite is inflicted. Horse-flies are most active in
hot weather, mostly in summer and autumn during the daylight hours. Most species
also prefer a wet climate, which makes it easier for them to breed. Eggs are generally
laid on stones close to water or on plant stems or leaves. On hatching, the larva fall
Figure 2. Giardia lamblia into water or moist earth, feeding voraciously on invertebrates, such as snails and
16 149

Figure 43. Phlebotomus papatasii
Black Fly
A Black Fly (sometimes called a Buffalo Gnat or Turkey Gnat) is any member
of the family Simuliidae of the Culicomorpha infraorder. There are over 1800 known
species of Black Flies (of which 11 are extinct). Like mosquitoes, to which they are
related, most Black Flies gain nourishment by sucking the blood of other animals.
They are usually small, black or gray, with short legs and antennae. They are a com-
mon nuisance for humans, and many U.S. states have programs to supress the Black
Fly population. They spread several diseases, including river blindness in Africa.
Horse-Flies
Among the world’s largest flies are the horse-flies (family Tabanidae). Although
not all the species in this family bite, these large, hairy flies are most often known as
pests because of the painful bites many species can inflict on animals and humans
(Fig. 44). They occur worldwide, being absent only at extreme northern and southern
latitudes. Flies of this type are among those known sometimes as «gadflies». A type
of insect, horse-flies are classified in the fly order Diptera. There are approximately
3,000 species of horse-flies known worldwide, 350 of which are found in North
America. At least three subfamilies are recognised:
! Chrysopsinae
! Pangoniinae
! Tabaninae
! The genus Zophina is of uncertain placement, though it has been classified
among the Pangoniinae.
The two best-known types are the common horse-flies, genus Tabanus, and the
148
Morphology of cysts
The cysts of G. lamblia are 8-12 mm in length and are ellipsoid in shape. They
contain 4 nuclei, which tend not to be obvious. Longitudinal fibrils consisting of the
remains of axonemes and parabasal bodies may also be seen. Cysts may appear to
shrink from the cell wall. The cysts are infective as soon as they are passed.
Clinical Disease
Giardia lamblia colonizes the small intestine where the trophozoites adhere to
the mucosal surface by means of their sucking disc. Cysts are produced as the parasites
descend the intestinal tract although trophozoites can be passed in the faeces in
severe infections. G. lamblia is transmitted through ingestion of cysts in contaminated
water or food. Cysts can survive outside the body for several weeks under favourable
conditions. The main symptoms are abdominal pain, flatulence, and episodic diarrhoea
with steatorrhea and periodical soreness in severe cases. No blood or mucus is
normally seen. However 50% of G. lamblia infections are symptomless, although
severe infections may develop in immunocompromised hosts. What determines
susceptibility is poorly understood. After swallowing cysts for the first time, symptoms
commonly develop 2-6 weeks later.
Normally illness lasts for 1 to 2 weeks, but there are cases of chronic infections
lasting months to years. Chronic cases, both those with defined immune deficiencies
and those without, are difficult to treat. The disease mechanism is unknown, with
some investigators reporting that the organism produces a toxin while others are
unable to confirm its existence. The organism has been demonstrated inside host
cells in the duodenum, but most investigators think this is such an infrequent
occurrence that it is not responsible for disease symptoms. Mechanical obstruction
of the absorptive surface of the intestine has been proposed as a possible pathogenic
mechanism, as has a synergistic relationship with some of the intestinal flora.
Thus, no matter what it looks like, stream water should be treated before drinking.
Boiling will kill Giardia cysts, and there are commercially available filters that will
remove the cysts from water.
Laboratory Diagnosis
Cysts can be found by examination of the deposit of a formol-ether concentrate
of a stool preparation. The oval cysts with thick walls serve as characteristic features
for these organisms. The flagella disintegrate and form a central “streak” which
becomes visible when stained with iodine or MIF (merthiolate-iodine-formaldehyde).
Cysts may be excreted intermittently; therefore it is important to examine more than
one stool. Stools are usually passed 3-8 times / day and are usually pale, offensive,
rather bulky and accompanied by much flatus.
Trophozoites are found by examination of saline wet preparations of fresh,
diarrhoeic stool, duodenal or jejunal aspirate or in a permanently stained faecal
preparation.
Trophozoites can also be found in the jejunal aspirate. These can be recovered
by the String Test or Enterotest capsule and the material examined microscopically
for motile trophozoites.
17
 Trophozoites and cysts can be found to be scarce in chronic infections. Serologi-
cal methods of diagnosis are proving to be useful as means of diagnosis. An ELISA
to detect IgM in serum provides evidence of a current infection. A polyclonal
antigen-capture ELISA can be used to demonstrate submicroscopic infections in
faeces and an IgA-based ELISA will detect specific antibodies in saliva.
Several strains of G. lamblia have been isolated and described through analysis
of their proteins and DNA; type of strain, however, is not consistently associated
with disease severity. Different individuals show various degrees of symptoms when
infected with the same strain, and the symptoms of an individual may vary during
the course of the disease.
Infectious Dose – Ingestion of one or more cysts may cause disease, as contrasted
to most bacterial illnesses where hundreds to thousands of organisms must be
consumed to produce illness.
Trichomonas vaginalis
(trichomoniasis, «trich» or «trick»)
Trichomonas vaginalis is a sexually transmitted disease (STD), although
transmission by other routes (such as soiled towels) has been documented. There is
no cyst in the life cycle, so transmission is via the trophozoite stage. Most people
infected with trichomoniasis are asymptomatic. Symptomatic infections are
characterized by a white discharge from the genital tract and itching. Diagnosis
depends on finding trophozoites in secretions of the genital tract from men or women.
In cases where the numbers of organisms are very low, the trophozoites can be cultured
to increase their numbers.
Trichomonas vaginalis, Pentatrichomonas hominis,
Enteromonas hominis,
and Retortamonas intestinalis
Classification
Protozoa. Phylum Sarcomastigophora. Flagellates.
Disease
Vaginal trichomoniasis in females and occasional symptomatology associated
with infections of the prostate and epididymis of males (T. vaginalis); P. hominis, E.
hominis, and R intestinalis are non pathogenic (Fig. 3).
Geographic Distribution. Worldwide.
Location in Host
Mucosal surface of the vagina, the prostate gland and seminal vesicle
(T. vaginalis), and intestinal tract (P. hominis, E. hominis, and R.. intestinalis).
Morphology. Trophozoites
Trophozoites of T. vaginalis are pyriform in shape, 7-30 µm long, with a width
of 6-15 µm. Living trophozoites have jerky, nondirectional movement. They have
18
Ivermectin suppresses microfilaria production but its overall effectiveness remains
untried and elephantiasis can be treated surgically.
Culex mosquitoes
Class: Insecta
Order: Diptera
Genus: Culex
General Characteristics: Culex are distinguished by their lack of colouration
and feature. The thorax, abdomen, legs and wings are often covered with brown-
black scales giving a generally dark appearance. The abdomen may occasionally
also have white scales arranged in segments. Culex breeds mainly in aquatic habitats,
often in areas containing large quantities of organic waste.
Life Cycle: Female Culex lay dark brown eggs in characteristic clumps of
approximately 300 eggs. As mentioned these eggs are often found in organic waste
deposits or polluted waters. Culex larvae have a long and narrow siphon with more
than one pair of subventral tufts.
Disease: Culex mosquitoes are vectors of Bancroftian filariasis throughout Africa,
but most importantly arboviruses such as Japanese encephalitis. Encephalitis occurs
throughout the world, with Culex acting as an important vector for spread and
infection. Culex mosquitoes are similar to the Culicine and Aede mosquitoes, but
prefer to bite at night and breed in organic refuse.
Control and Treatment: Culex mosquitoes are most easily controlled by
improving sanitation and removing static water sources from the affected area. In
general the most effective control for Culex mosquitoes are also repellents and fine
screening or netting. Treatment with insecticides will also serve to reduce the vector
population, but increased problems are encountered with Culicine mosquitoes because
they also feed during the daytime. If filarial infection occurs treatment with
Diethylcarbamazine (DEC) will kill microfilaria. Ivermectin suppresses microfilaria
production but its overall effectiveness remains untried and elephantiasis can be
treated surgically.
Phlebotomus
Phlebotomus is a genus of flies, or diptera, that generally includes «sand flies»
(Fig. 43). In the Old World, Phlebotomus sand flies are primarily responsible for the
transmission of leishmaniasis, an important parasitic disease. Leishmaniasis is
generally transmitted in the New World by sand flies of the genus Lutzomyia. The
parasite itself is a species of the genus leishmania, a protozoan. The disease normally
finds a mammalian reservoir in rodents and other small animals such as canids and
hyraxes. The sand fly carries the leishmania protozoa from infected animals after
feeding, thus transmitting the disease.
147
 Aedes mosquitoes

Class: Insecta
Order: Diptera
Genus: Aedes
General Characteristics: Aedes can generally be distinguished by patterns of
black and silvery scales present on the abdomen and thorax. The legs appear to have
black and white rings along their length. The wings are generally covered with black
scales. Aedes breed in marshes and other wetland areas and have a worldwide
distribution.
Life Cycle: Female Aedes lay eggs on damp areas such mud, detritus, clay and
rock. The eggs are very robust and can survive desiccation and other environmental
pressures. The eggs hatch in waves depending on the environmental cues. Aedes
larvae have a stout barrel shaped siphon with one pair of subventral tufts. There are
three pairs of setae on the ventral brush, and large setae are not present on the
abdominal segments.
Disease: Aedes are vectors of Bancroftian filariasis and arboviruses such as yellow
fever and dengue. Wuchereria bancrofti is the main cause of «elephantiasis»
(Bancroftian filariasis) and the most widely distributed filarial parasite of Man. The
adults live in the lymphatic system, and can survive for 30 years or more. They
copulate and generate a pre-larval form, the microfilaria. Both the adults and the
microfilaria may play a role in generating the symptoms and signs. Microfilaria
measure 240-300 mm in length by 7-10 mm in width. They are sheathed (derived
from ovum membrane) and nuclei terminate 15-20 microns proximal to the pointed
tail. There are fewer, more distinct nuclei than in other species and there are less
body curves. Adult worms are slender and white (males 4 cm; females stout and 10
cm in length).
Initial infection with Wuchereria is usually asymptomatic. There may be
recurrence of attacks of «cellulitis» affecting the limbs, breast, scrotum or elsewhere.
Infection is associated with fever, lymphangitis, lymphadenopathy and occasionally
abscess formation. These initially settle but later on the tissues eventually become
oedematous and hypertrophied. Further effects may include scrotal involvement and
hydrocoele, which can lead to scrotal enlargement and lymph scrotum. This is
«elephantiasis» and is associated with dermal hypertrophy, verrucous changes and
the rupture of lymph varices into various sites.
Yellow fever and dengue haemorrhagic fever are serious viral infections spread
by the Aedes mosquito. Dengue is now the most important mosquito borne virus,
with global infection increasing. Figure 3. Trichomonas vaginalis
Control and Treatment: In general the most effective control for Culicine
mosquitoes are also repellents and fine screening or netting. Treatment with four anteriorly directed flagella and a fifth one directed posteriorly along the outer
insecticides will also serve to reduce the vector population, but increased problems margin of the undulating membrane; the latter extends only half the distance to the
are encountered with Culicines because they also feed during the daytime. If filarial posterior end of the body. The nucleus is usually elongated in the anterior portion of
infection occurs treatment with Diethylcarbamazine (DEC) will kill microfilaria. the organism. The interior of the nucleus contains many chromatin granules and a
146 19
small karyosome. The cytoplasm contains many dark-staining granules
(hydrogenosomes), but these are not readily seen in Giemsa-stained specimens. A
rodlike, pointed axostyle protrudes from the posterior end of the body.
Pentatrichomonas hominis trophozoites are usually 6-14 µm long but some may be
up to 20 µm long. They are morphologically similar to T. vaginalis except that the
posteriorly directed flagellum that forms the outer edge of the undulating membrane
projects beyond the posterior end as a free flagellum. The axostyle is a slender rod
extending from the anterior end through the middle of the body and projects from
the posterior end. The anteriorly placed nucleus contains a small karyosome.
Hydrogenosomes are readily visible in trichrome-stained organisms.
Trophozoites of Enteromonas are usually 6-8 µm long but may be somewhat
shorter or longer. They have three anteriorly directed flagella and a fourth one that
extends posteriorly beyond the end of the body. They move in a rapid, jerky fashion.
Their nucleus is near the anterior end and has a large, central karyosome. With fixation,
trophozoites are spherical or ellipsoidal and are somewhat smaller in size.
Trophozoites of Retortamonas are ovoid or pyriform, 4-10 µm long by 3-7.5 µm
wide, and have two flagella, one directed anteriorly and the other extending
posteriorly. A cytostome is present at the anterior end; it extends posteriorly for
nearly half the length of the organism and is bordered by a fibril. The nucleus is
spherical, located in the anterior end, and it contains a small karyosome. There is a
fine layer of peripheral chromatin.
Cysts
Trichomonas vaginalis and P. hominis lack a cyst stage. Cysts of E. hominis are
small and inconspicuous, usually 4-8 µm long by 3-5 µm in width, and ellipsoidal in
shape. Cysts will contain one, two, or four nuclei but binucleate forms predominate.
When two nuclei are present, they frequently are at opposite ends; in those cysts with
four nuclei, they are often paired at opposite ends. These nuclei have large, central
karyosomes surrounded by a clear space; the nuclear membrane lacks peripheral
chromatin. Cysts of Retortamonas are ovoid or pyriform in shape and measure 4-7 µm
long by 3.5-4.5 µm in width. Mature cysts are uninucleate, have a compact central
karyosome, and varying amounts of peripheral chromatin. The fibril associated with
the cytostome in the trophozoite may be seen in the cyst, often near the nucleus.
Life Cycle
Direct transmission of the trophozoite during sexual intercourse (T. vaginalis)
and presumably by ingestion of the trophozoite stage {P. hominis) or the cyst stage
(Enteromonas and Retortamonas).
Diagnosis
The demonstration of trichomonads in vaginal secretions, scrapings, urethral
discharge and sedimented urine from women, and in prostatic secretions and
sedimented urine from men provide definitive diagnosis of T. vaginalis infection. In
women, material for examination is best recovered by using a platinum loop or cotton-
tipped swab applied to the fornices of the vagina. A sample of the scrapings from the
loop is placed directly in a drop of saline solution on a slide, a cover glass is applied,
20
Infection is associated with fever, lymphangitis, lymphadenopathy and occasionally
abscess formation. These initially settle but later on the tissues eventually become
oedematous and hypertrophied. Further effects may include scrotal involvement and
hydrocoele, which can lead to scrotal enlargement and lymph scrotum. This is
«elephantiasis» and is associated with dermal hypertrophy, verrucous changes and
the rupture of lymph varices into various sites.
Brugian (Malayan) filariasis is less widespread, less common and less serious
than its Bancroftian counterpart. The life cycle is identical to that of Wuchereria
bancrofti with Brugia malayi limited to Asia and B.timori restricted to Indonesia.
Infection results in lymphadenopathy involving most frequently the inguinal area,
lymphoedema normally below the knee, eosinophilia, and in rare cases chyluria.
Treatments and Control
Malaria: If the infective species is not known, or the infection is known to be
mixed, initial treatment should be with quinine, mefloquine or rarely halofantrine.
Falciparum (malignant) malaria is often resistant to chloroquine and should be
treated with quinine, mefloquine, halofantrine, quinidine or pyrimethamine-
sulphadoxine. Benign malaria (P. vivax) should be treated with chloroquine al-
though resistance has been reported from New Guinea. Malarial prophylaxis is
relative and not absolute.
The UK Consensus Group on Malaria Prophylaxis (1997) recommend mefloquine
for UK travellers to West, Central and East Africa for periods of greater than 2
weeks and for travellers to specific areas within south-east Asia: prophylaxis should
be commenced 2 weeks before departure. Doxycycline can be used in older children
and adults who cannot tolerate mefloquine.
Prevention is most dependent upon coverage of exposed skin and the use of
insect repellent, mosquito nets impregnated with permethrin and correct
prophylaxis. The vector may be controlled by water clearance programs, house
spraying (DDT) and destruction of breeding areas. Drug resistance to DDT and
ethical resistance to its use have limited its effectiveness. Natural immunity
involves both antibody and cell-mediated systems and appears to require frequent
boosting; antigens from different stages of the parasite’s life cycle will be
important in vaccine development.
Filariasis
Diethylcarbamazine (DEC) kills microfilaria. Ivermectin suppresses microfilaria
production but its overall effectiveness remains untried and elephantiasis can be
treated surgically. Control measures comprise draining of mosquito breeding sites
and killing larvae. Many mosquitoes are resistant to insecticides but mosquito
repellents and nets are effective. The infective pool may be reduced by periodic
mass treatment with DEC. Brugia malayi is more susceptible to diethylcarbamazine
(DEC) than is Wuchereria bancrofti. Anopheline larvae may be suffocated in their
breeding sites but culicine larvae (Mansonia sp.) derive oxygen from plants and
are not amenable to such measures. Control depends upon the use of mosquito
nets and periodic mass treatment.
145
lytic or due to toxic marrow suppression. Splenomegaly occurs in all malaria: it
may be acute or chronic (+/- hypersplenism). Jaundice may be haemolytic and/or
hepatic (only P. falciparum). In addition, there may be headache, myalgia, arthral-
gia, diarrhoea and vomiting.
Plasmodium falciparum is the most virulent form (invades mature and immature
RBCs) and is often fatal if untreated. Blood schizogony takes place in deep capillaries
and micro-circulatory failure can occur in individuals with little immunity to malaria.
It does not relapse but recrudescence may occur. The time between paroxysms is 48
hours but fever may last for 24-36 hours. Very rapid progression and complications
include diarrhoea and vomiting; delirium; coma; convulsions; renal failure, including
haemoglobinuria (blackwater fever); jaundice; pulmonary oedema; hypoglycaemia
and abortion. Cerebral malaria often results in delirium, disorientation, stupor, coma,
convulsions and death.
P. vivax / ovale exhibit 48 hours between paroxysms; relapses may occur up to 8
years after primary infection and only infects immature RBCs of those with Duffy
blood group. Plasmodium malariae generally results in72 hours between paroxysms,
only infects older RBCs, and recrudescence may occur decades after primary infection.
The global malaria situation is serious and becoming worse: 300-500 million
clinical cases occur annually. 1.5-2.7 million people die of malaria each year
with approximately one million deaths among children under five years of age
are attributed to malaria alone or in combination with other diseases. Countries
in tropical Africa account for more than 90% of the total malaria incidence and
the great majority of malaria deaths (WHO data). The death toll of African
children with malaria is expected to double by 2010, conceivably reaching 4
million deaths per year. Many factors influence the epidemiology of this disease
including: breeding habits of the various mosquito vectors; agricultural practices;
economic conditions; industrialisation and pesticide use. Increasing air-traffic
from malaria endemic areas has led to the possibility of malaria developing in
non-endemic areas where the mosquito vector has been imported onboard aircraft.
Filariasis
Anopheline mosquitoes also transmit the filarial worms Wuchereria bancrofti,
Brugia malayi and Brugia timori. Wuchereria bancrofti is the main cause of
«elephantiasis» (Bancroftian filariasis) and the most widely distributed filarial parasite
of man. The adults live in the lymphatic system, and can survive for 30 years or
more. Once they have mated they produce a pre-larval form, the microfilaria. Both
the adults and the microfilaria may play a role in generating the symptoms and signs.
Microfilaria measure 240-300mm in length by 7-10mm in width. They are sheathed
(derived from ovum membrane) and nuclei terminate 15-20mm proximal to the
pointed tail. There are fewer, more distinct nuclei than in other species and there are
less body curves. Adult worms are slender and white (males 4 cm; females stout and
10 cm in length).
Initial infection with Wuchereria is usually asymptomatic. There may be recur-
rence of attacks of «cellulitis» affecting the limbs, breast, scrotum or elsewhere.
144
and then the sample is examined by light, darkfield, or phase contrast microscopy
for the presence of typical motile trophozoites. Organisms must be distinguished
from squamous epithelial cells and polymorphonuclear leukocytes, which are also
collected in the sampling procedure. Material taken by cotton swab should be agitated
in a few drops of saline solution in a small test tube and the suspension then examined
in a microscope slicie preparation. Fluorescence microscopy using vital dyes such
as acridine orange has also been used but to no apparent advantage over direct wet
mount preparations. Giemsa-stained smears have been used when it is not feasible
to examine wet mounts immediately. Trichomonads can be found in Papanicolaou-
stained smears, but the morphology of the organisms is often altered by the staining
procedure, making identification more difficult. Various culture media, including
Diamond’s medium, can be inoculated with material from the vagina, incubated
anaerobically, and examined several days after inoculation for the presence of motile
trophozoites. Commercially available culture systems have been developed that are
useful for diagnosis of this infection. Also, urine samples can be sedimented and
examined for the presence of organisms.
The isolation of T. vaginalis from men is difficult and is best accomplished by
examination of urine or prostatic secretions. Scrapings of urethral mucosa with a
platinum loop and examination of this material as a wet mount preparation, or by
inoculation of the material into culture media, are the usual procedures employed. If
urine is examined, the sample should be taken at the first opportunity in the morning,
preferably after prostatic massage; the sediment can be examined directly for the
presence of organisms or it can be placed into culture media.
The diagnosis of P. hominis, Enteromonas, and Retortamonas trophozoites in
feces is best accomplished by direct wet mount preparations that reveal the typical
jerky movement of these organisms. All of these organisms can be found in stained
fecal smears; however, their affinities for stain are inconsistent.
Diagnostic Problems
Trophozoites of T. vaginalis frequently are difficult to find in wet mount
preparations; however, the use of culture methods will increase the possibility of
detecting an infection. A variety of staining methods have been used for demonstrating
organisms. In Papanicolaou-stained smears, however, organisms are frequently
distorted and difficult to identify; if a Gram stain is used, the organisms stain similarly
to polymorphonuclear leukocytes and can be misidentified. In trichrome-stained fecal
smears, P. hominis rarely stains well and is easily overlooked. The small size of both
Enteromonas and Retortamonas results in these infections being rarely diagnosed
or reported. The small size of E. hominis cysts leads to their being confused with
Endolimax nana cysts.
Comment
Trichomonas vaginalis infection is an extremely important sexually transmitted
disease throughout the world. In the United States, more than 1 to 2 million cases
occur annually, and it is believed that on a worldwide basis there may be 100 to 200
million cases each year.
21
 Trichomonas hominis
Introduction
This flagellate is cosmopolitan in its distribution. It is thought to be non-pathogenic
although it has been associated with diarrhoeic stools. It is the most commonly found
flagellate next to G. lamblia and D. fragilis. Found in a wide host range including
non-human primates, cats, dogs and various rodents.
Morphology of trophozoites
Trichomonas hominis do not have a cystic stage. The trophozoites measure from 5-15
mm in length by 7-10 mm in width. The shape is pyriform and has an axostyle which runs
from the nucleus down the centre of the body and extends from the end of the body. They
also possess an undulating membrane, which extends the entire length of the body and
projects from the body like a free flagellum (this feature distinguishes it from other
trichomonads). The characteristic number of flagella is five; there is some deviation from
this number. They also have a single nucleus at the anterior end. Trichomonads swim with
a characteristic wobbly movement, which makes them unmistakable during diagnosis.
Laboratory Diagnosis
In a fresh stool, the flagellates move very rapidly in a jerky, non-directional manner.
The axostyle and undulating membrane are diagnostic. The flagellates are difficult to
stain, however, the axostyle can be seen on a stained preparation and is diagnostic.
Table 1
Differential morphology of flagellates found in stool samples of humans.
* Not a normal feature for identifying species in routine stool samples
22
General Characteristics: Anopheles mosquitoes are characterised by dark
and pale scale blocks arranged on their wings. They have palps that are of equal
length to the proboscis, which appear terminally clubbed in males. Anopheles
always rest at an angle when standing on surfaces and preferring to feed at twilight
or night. Breeding sites are varied but Anopheles prefer unpolluted fresh or
saltwater.
Life Cycle: Anopheles lay 50 to 200 dark colour eggs in aquatic environments,
and hatch in several days to several weeks depending on the external temperature.
Anopheles larvae have a dark brown head and 6-7 anterior segments covered with
dorsal palmate hairs. Accessory tergal plates are present on the dorsal side of
segments 1-10 and two sets of anal papillae emerge from the last abdominal
segment.
There are four larval instars that survive by filter feeding and breathing oxygen
through their spiracles. Anopheles larvae occur throughout many different habitats
including both permanent marshes and swamps, and temporary locations such as
pots filled with water. In general Anopheles prefer to inhabit clean habitats. The
larval period lasts about a week, but may be extended depending on the
environmental conditions. The pupa is comma shaped with a set of trumpet shaped
breathing tubes. The abdomen is covered with setae, and segments 2-7 have distinct
spines. The pupal period may last a few days to weeks depending on the temperature.
Disease
Malaria: Anopheles are vectors of malaria, Bancroftian and Brugian filariasis
and of multiple arboviruses (dengue fever; yellow fever; encephalitides and
haemorrhagic fevers). Malaria is caused by Plasmodium falciparum, P. vivax, P.
malariae and P. ovale. Transmission of the disease occurs in virtually all of
tropical Africa, Central and South America, and the Middle and Far East. South
East Asia is a particular problem due to multiple drug resistance. P. falciparum
is found in Africa and other tropical countries as well as in subtropics. P. malariae
has a low prevalence in both tropics and subtropics. P .vivax is the most
widespread in temperate regions and subtropics but may also be found in the
tropics. P. ovale has a low prevalence in West Africa. In Africa alone, 370 million
people live in endemic areas.
P.vivax causes benign tertian malaria (43% of cases) and P. falciparum results
in malignant tertian or sub-tertian malaria and pernicious malaria (50% of cases).
P .ovale (mild tertian malaria, 1% of cases) and P. malariae (quartan malaria, 7%
of cases) contribute a small percentage of malarial cases.
Clinical features including fever and chills are due to the host inflammatory
response and are associated with rupture of erythrocytic schizonts. Fever presents
in three stages – a) Cold: rigors and fever lasting 15 minutes to 1 hour; b) Hot: the
patient is flushed with tachycardia and is pyrexial (40C) for 2-6 hours; c) Sweating:
the temperature falls (over 2-4 hours). Each paroxysm lasts 8-12 hours in total. All
erythrocytes containing a trophozoite will be destroyed within 48-72 hours. Peri-
odic fever often takes more than 7 days to develop, and anaemia can be haemo-
143

Figure 42. Comparison of malarial and nonmalarial mosquitoes. 1. Eggs. 2. Respiratory
foramen in larvae. 3. Respiratory tubes in larvae. 4. Antennae. 5. Bristle;like antennae.
6. Sucking tubes. 7. Eyes. 8. Thoracic part. 9. Abdomen.
males are pilose (few spidery hairs). The male Anopheline palps are long and clubbed;
those of the male Culicine are long but not clubbed. The female Anopheline palps
are long; the female Culicine are short. If a mosquito is incorrectly sexed, a female
Anopheline may be confused with a male Culicine (Fig. 42).
Mosquitoes
Class: Insecta
Order: Dipthera
Genus: Anopheles
142
Trypanosoma species
Introduction
Trypanosomes are haemoflagellates and three species of the genus Trypanosoma
are responsible for disease in humans such as sleeping sickness.
Trypanosomes occur in the blood of the majority of vertebrate animals. The life
cycle involves intermediate host, which usually is an insect. Many species of
trypanosomes can live in harmony with their hosts producing no pathogenic effect,
but the best known species are those that are pathogenic to their definitive hosts. The
disease in caused by the pathogenic types is called, trypanosomiasis.
Salivarian trypanosomes
Trypanosoma brucei rhodesiense and Trypanosoma brucei gambiense – The
metacyclic trypanosomes are found in the proboscis of the insect vector – infection
is therefore inoculative. The above are the aetiological agents of African
trypanosomiasis, it is a zoonotic species in the fact that it multiplies in the blood of
a range of animals, domestic and wild animals as well a man.
Trypanosoma brucei rhodesiense causes an acute form of sleeping sickness in
East Africa, while T. b. gambiense causes chronic sleeping sickness in West Africa.
These are known as salivarian trypanosomes as they complete their development
in the salivary system (anterior portion of the vector). Transmission takes place by
innoculation of the metacyclic stage.
Stercorarian trypanosomes
Trypanosoma cruzi – The metacyclic trypanosomes occupy a posterior position
in the gut of the insect vector and are passed out in the faeces – infection is therefore
contaminative. This is the aetiological agent of South American trypanosomiasis.
These trypanosomes are known as stercocarian as they complete their development
in the posterior region of the vector, so that the infective forms appear in the insects’
faeces. Hosts are infected by the contaminative route.
African Trypanosomiasis
Life Cycle
Transmission from one vertebrate to another is carried out by blood-sucking
invertebrates, usually an insect. The vector for African Trypanosomes is the Tse tse
fly, Glossina, and the species which cause the disease are T. b. gambiense (Fig. 4)
and T. b. rhodesiense.
Metacyclic (infective) trypomastigotes are inoculated through the skin when a
tsetse fly takes a blood meal. The parasites develop into long slender trypomastigotes,
which multiply at the site of inoculation where ulceration occurs. The trypanosomes
23
continue to develop and then may invade the lymphatic tissues, the heart, various
organs and in later stages, the central nervous system. Trypomastigotes are taken up
by the tsetse fly (male and female) during a blood meal. The parasites develop in the
midgut of the fly where they multiply. 2-3 weeks later the trypomastigotes move to
the salivary glands transforming from epimastigotes into metacyclic (infective)
trypomastigotes. The tsetse fly remains infective for life i.e. about 3 months.
The mode of transmission mentioned above, metacyclic transmission, requires to
be separated from mechanical transmission, a process in which trypanosomes survive,
for a short time, on and about mouth parts of an insect and are inoculated into a new
host when the vector bites again, without undergoing any developmental cycle.
Metacyclic transmission requires a lapse of time to allow the trypanosomes to
reach an infective stage by a particular developmental sequence in the vector, usually
a period of several days.
Morphology
The parasite is an elongated cell with single nucleus, which usually lies near the
centre of the cell. Each cell bears a single flagellum, which appears to arise from a
small granule – the kinetoplast. The kinetoplast is a specialised part of the
mitochondria and contains DNA. The length and position of the trypanosome’s
flagellum is variable. In trypanosomes from the blood of a host the flagellum originates
near the posterior end of the cell and passes forward over the cell surface, its sheath
is expanded and forms a wavy flange called an undulating membrane.
Development is characterised by the occurrence of three types of blood forms
(polymorphic), these are:
1) Slender forms: long and thin, about 29mm long, free flagellum.
2) Stumpy forms: thick and short, average length 18mm, typically no free
flagellum, but a short one may be present.
3) Intermediate forms: about 23mm long with a moderately thick body and a
free flagellum of medium length.
Clinical disease
The early stages of African trypanosomiasis may be asymptomatic and there is a
low grade parasitiaemia. This period may last for several weeks to several months.
The disease may terminate untreated at this stage or go on to invade the lymph
glands. Invasion of the lymph glands is usually accompanied by a high irregular
fever with shivering, sweating and an increased pulse rate. The lymph glands near
the bite often become swollen, in T. b. gambiense the glands at the back of the neck
and T. b. rhodesiense usually the glands under the jaw are affected (Winterbottoms
sign). As the disease progresses oedema of the eyelids, face and sleeplessness are
features along with increasing lethargy and listlessness.
Trypanosomes may invade the central nervous system giving symptoms of
meningoencephalitis, confusion, apathy, excessive sleeping and incontinence. At this
stage, the cerebro-spinal fluid (CSF) usually contains mononuclear cells and a few
trypanosomes may be detected. If untreated, character changes, mental deterioration
and coma develop, finally resulting in death. Such signs are more commonly seen
24
the transmission of plague. Infection is often spread by the bite alone, but can also poten-
tial be transmitted through fecal abrasion. Tunga penetrans does not transmit disease to
humans, but females will burrow into host skin. The pinpoint lesion enlarges to pea-size
within 2 weeks necessitating removal of the gravid female using a pin, a needle or a
sliver of bamboo. This may potentially lead to a secondary bacterial infection.
Control and Treatment: Control of fleas is generally mediated through
insecticidal powders and aerosols. If outbreaks of murine typhus or plague occur
steps to control the rodent populations in the affected area may be employed.
Diptera
Diptera are insects in which the hind wings are reduced to halteres. The study of
the diptera is called dipterology.
Synonums and common names: Flies, mosquitoes, gnats and midges are the
species in this order. In compound names containing «fly» for members of this order,
the name is written as two words as in «crane fly». For insects that are members of
other orders the name is written as a single word as in “butterfly”.
Subdivision: About 85,000 species of insect have only two wings and are classified
as members of Diptera, There are two generally accepted sub-orders of Diptera. The
Nematocera are usually recognized by their elongated bodies and feathery antennae as
represented by mosquitoes and crane flies. The Brachycera tend to have a more roundly
proportioned body and very short antennae. Beyond that, considerable revision in the
taxonomy of the flies has taken place since the introduction of modern cladistic
techniques, and much remains uncertain. The secondary ranks between the sub-orders
and the families are more out of practical considerations than out of any strict respect
for phylogenetic classifications. (Modern cladists tend to spurn the use of any rank
names.) Several of the classifications used now in this article remain paraphyletic
groupings; this is particularly notable in the Orthorrapha.
Mosquitoes
Class: Insecta
Order: Dipthera
Genus: Anopheles, Aedes, Culex
Introduction: Mosquitoes are small with a clearly demarcated body and very
long slender legs. The head contains a large pair of kidney shaped compound eyes,
a pair of antennae, and a single long proboscis for feeding. The thorax, abdomen and
wings are often covered with scales. Differential colouration and pattern of these
scales provides a means of visually distinguishing species. The large wings are folded
over the segmented abdomen, which generally appears brown-black and slender but
turns a bright red and swells following feeding.
Mosquitoes may be classified as Anopheline (Anopheles) or Culicine (Aedes,
Culex). The antennae of male mosquitoes are plumose (many feathery hairs); fe-
141
 Figure 41. Fleas

eight months depending on the temperature, humidity, food, and species. Normally after a
blood meal, the female flea lays about 15 to eggs per day – up to 600 in its lifetime – usually
on the host (dogs, cats, rats, rabbits, mice, squirrels, chipmunks, raccoons, opossums, foxes,
chickens, humans, etc.). Eggs loosely laid in the hair coat drop out almost anywhere, espe-
cially where the host rests, sleeps or nests (rugs, carpets, upholstered furniture, cat or dog
boxes, kennels, sand boxes, etc.). Eggs hatch between two days to two weeks into larvae
found indoors in and along floor cracks, crevices, along baseboards, under rug edges and in
furniture or beds. Outdoor development occurs in sandy gravel soils (moist sand boxes, dirt
crawlspace under the house, under shrubs, etc.) where the host may rest or sleep. Sand and
gravel are very suitable for larval development, which is the reason fleas are erroneously
called «sand fleas.» Larvae are blind, avoid light, pass through three larval instars and take
a week to several months to develop. Their food consists of digested blood from adult flea
feces, dead skin, hair, feathers, and other organic debris; larvae do not suck blood. Pupae
mature to adulthood within a silken cocoon woven by the larva to which pet hair, carpet
fiber, dust, grass cuttings, and other debris adheres. In about five to fourteen days, adult
fleas emerge or may remain resting in the cocoon until the detection of vibration (pet and
people movement), pressure (host animal lying down on them), heat, noise, or carbon
dioxide (meaning a potential blood source is near). Most fleas overwinter in the larval or
pupal stage with survival and growth best during warm, moist winters and spring. Flea bites
can be treated with Calamine Lotion or 0.5-1% conc. hydrocortisone cream. Lufenuron is
a veterinary medicine that attacks the larval fleas ability to produce chitin.
Disease: Fleas are a general nuisance, often biting humans on exposed surfaces
resulting in discomfort. Flea-bites induce pruritic papular urticaria commonly on the
unprotected lower leg of women and all over the body of children who have intimate
animal contact; a generalised allergic response may occur.
Certain fleas, notably the rat fleas, spread plague (Yersinia pestis) and murine typhus
(Rickettsia typhi), and serve as intermediate hosts for species of tapeworm (Hymenolepis
sp.). Cat and dog fleas serve as intermediate hosts for another common tapeworm (Dipy-
lidium caninum), which can be spread to humans, especially children with exposure to
pet animals. Pulex irritans is not a major vector of disease but may play a minor role in Figure 4. Trypanosoma gambiense
140 25
with gambiense than in rhodesiense in which patients often die before these symp-
toms develop fully.
Laboratory diagnosis of African trypanosomiasis
Laboratory diagnosis of African trypanosomiasis is by:
1. Examination of blood for the parasites
2. Examination of aspirates from enlarged lymph glands for the parasites
3. Examination of the CSF for the parasite
4. Detection of trypanosomal antibodies in the serum
1. Examination of blood
a) Thick and thin blood films blood film (thick and thin blood films are made
and stained with Fields stain and examined as for malaria parasites).
b) Triple centrifugation technique.
This method is carried out as follows:
(²) 5 to 10 ml of citrated blood is centrifuged at 2000 rpm for 5 minutes to pack
the red blood cells.
(²²) The plasma and white cell layer are removed by a Pasteur pipette and
transferred to a clean centrifuge tube.
(²²²) This is centrifuged for a short time in order to deposit any red blood cells
carried over.
(²V) The supernatant fluid is removed by pipette to a clean tube.
(V) This is centrifuged at 5000 rpm for 10 minutes.
(V²) The supernatant fluid is removed with a pipette and discarded.
(V²²) The deposit is examined microscopically for trypanosomes.
c) Miniature anion-exchange centrifugation technique (ref. Transactions Royal
Society of
Tropical Medicine and Hygiene. 1979. 73. 312-317)
Heparinised blood is passed through an anion exchange column. As the blood
travels down the column the red cells are adsorbed while the less strongly charged
trypanosomes are washed through with saline. The eluate is centrifuged and examined
microscopically for motile trypanosomes.
d) Buffy coat examination.
Trypanosomes are centrifuged in a microhaematoctit tube for 5 minutes. Parasites
can be seen microscopically at the junction of the packed red cells and plasma.
2. Examination of lymph gland aspirates
The aspirate can be examined microscopically by making a wet preparation, or if
there is not much material, it can be allowed to dry on a slide and then stained with
either rapid Field’s stain or with Giemsa and examined microscopically.
3. Examination of CSF
In the late stages of African trypanosomiasis, trypanosomes may be found in the
CSF together with IgM – containing morula (Mott) cells, lymphocytes and other
26
Head lice are normally spread by close contact but can also be spread by sharing clothes.
Treatment: The most common Western treatment is with chemical insecticides such
as pyrethrin, however there is increasing controversy over possible toxic side effects. For
more information as well as alternate treatments, see Treatment of human head lice.
Crab Lice
Class: Insecta
Order: Phithiraptera
Genus: Phthirus
Introduction: The crab louse is 1-2mm long and distinguished by a square,
undifferentiated body and massive claws on the two posterior sets of forelegs. These
claws are able to grasp both pubic and facial hair (including eyelashes), and allow
the louse to remain tightly bound to the host. They are spread mostly by sexual
contact, but may also be transmitted through fomites.
Life Cycle: The life cycle of Phthirus is very similar to Pediculus. Females lay
bundles of eggs on the coarse pubic hairs and dense facial hairs of humans. The crab
lice proceed through a cycle similar to the head and body lice, with the nymphal
stage proceeding several days longer. Phthirus are less active than Pediculus, but
similarly cannot survive for very long without a host and blood meals.
Disease: There appears to be very little evidence of disease transmission by
Pthirus, but have the ability to cause severe localised allergic reactions during
infestations.
Fleas
Fleas are external parasites, living by hematophagy off the blood of mammals
and birds.
Note: There is also a genus of Protozoa named Siphonaptera.
Some well known flea species include:
! Cat Flea (Ctenocephalides felis),
! Dog Flea (Ctenocephalides canis),
! Nothern rat flea (Nosopsyllus fasciatus),
! Oriental Rat Flea (Xenopsylla cheopis).
In most cases fleas are just a nuisance to their hosts, but some people and some
animals suffer allergic reactions to flea saliva resulting in rashes. Flea bites generally
result in the formation of a slightly-raised swollen itching spot with a single puncture
point at the center. However, fleas can transmit disease. One devastating example of
this was the bubonic plague, transmitted between rodents and humans. Murine typhus
(endemic typhus) fever, and in some cases tapeworms can also be transmitted by fleas.
Life cycle
Fleas pass through a complete life cycle consisting of egg, larva, pupa and adult
(Fig. 41). Completion of the life cycle from egg to adult varies from two weeks to
139

Figure 40. Lice
brownish or greyish, but almost always lighter colored. Eggs normally undergo a
7-9 day incubation before hatching as a baby nymph. Classically, a louse egg does not
become a «nit» until after it has completed its incubation stage, thus leaving a «nit». A
«nit» is either the empty shell remaining after the nymph has departed or the dead egg
that remains if incubation was not successful. Dead eggs will appear dark, or raisin-
like, as they dry out. «Nits» are usually found one-half inch or more away from the
scalp and are not considered a sign of an active infestation. There are three nymph
instar stages as the baby louse matures, with the louse shedding its exoskeleton at the
end of each stage, as it gets larger. The nymph stage typically lasts 10 to 12 days.
Whether a louse is male or female is not apparent until they are nearly mature. Fertili-
zation of eggs takes place once as the female reaches the mature stage. The female can
then lay 3-7 eggs each day for the next 28 to 30 days, her normal life span. There are
three main stages in the life of a head louse: the nit, the nymph, and the adult.
Nit: Nits are head lice eggs. They are hard to see and are found firmly attached
to the hair shaft. They are oval and usually yellow to white. Nits take about 1 week
to hatch.
Nymph: The nit hatches into a baby louse called a nymph. It looks like an adult
head louse, but is smaller. Nymphs mature into adults about 7 days after hatching.
To live, the nymph must feed on blood. It metamorphoses 3 times before it reaches
the adult stage.
Adult: Females lay nits (a few hundreds of eggs); they are usually larger than
males. To live, adult lice need to feed on blood. If the louse falls off a person, it dies
within 1-2 days.
Symptoms: The louse feeds on human blood, and the bite causes itching. Bites
can become secondarily infected; scratching may break the skin and help cause this
secondary infection. The most common symptom is itching of the scalp.
138
mononuclear cells. Once the CSF has been collected it must be examined as soon
as possible. The parasites are unable to survive for more than 15-20 minutes in CSF
once it has been removed. The parasites become inactive, are rapidly lysed and will
not therefore be detected. The CSF should be examined wet and spun down in a
sterile tube and a film made from the deposit. The film is then stained with rapid
Field’s or Giemsa and examined microscopically.
NB. It is impossible to distinguish between T. b. gambiense from T. b. rhodesiense
on a stained film, as the two subspecies, which infect man, are identical.
South American trypanosomiasis
Introduction
Trypanosoma cruzi occurs throughout South and Central America, especially in
Brazil, Argentina and Mexico causing the disease known as Chagas’ disease. It is
estimated that over 24 million people are infected with this species. It is a zoonotic
parasite with over 150 species of wild animals known to harbour the parasites, for
example opposums, dogs, rates, pigs and cats (Fig. 5).
It is transmitted to man by brightly coloured bugs belonging to the Reduviidae
family, subfamily Triatominae. All stages of these bugs are known to become infected.
The bugs live in the crack of the walls and vegetal roofs of the poorly maintained
houses, coming out at night to feed on the exposed parts of the host’s body.
Life Cycle
Metacyclic trypomastigotes are deposited in faeces on the skin as the triatomine
bug (reduviid bug) feeds. The bug usually bites round the edges of the mouth and
eyes. The trypomastigotes are either rubbed into the skin by scratching the irritated
area or penetrate the conjunctiva or membranes of the nose and mouth.
Trypomastigotes become amastigotes in localised reticulo endothelial cells and
multiply. The amastigotes develop into trypomastigotes, which are released into the
blood when the cell ruptures. No multiplication of the parasite takes place in the
blood in its trypomastigote stage. The trypomastigotes reach tissue cells especially
heart muscle, nerves, skeletal muscle and smooth muscle of the gastrointestinal system
by way of the blood and lymphatic system. The trypomastigotes become amastigotes
and multiply forming pseudocysts. Within the pseudocyst some amastigotes become
elongated and develop first into epimastigotes and then trypomastigotes. When the
cell ruptures the trypomastigotes are released into the blood and continue to circulate
whilst others invade further tissue cells. The life cycle completes when a triatomine
bug vector ingests circulating trypomastigotes. In the vector the trypomastigotes
transform and develop into epimastigotes, multiply by binary fission in the gut of the
bug. After about 10-15 days, metacyclic trypomastigotes are formed and can be
found in the hindgut of the bug.
Morphology
Trypanosoma cruzi has a single form (monomorphic), about 20mm in length,
and characteristically curved. The kinetoplast is large, considerably larger than the
27

Figure 5. Trypanosoma cruzi
Trypanosoma bruceii species already discussed. They sometimes appear as a bulge
at the posterior end. The flagellum is medium in length.
28
or less-developed areas can reduce their chances of being bitten by staying in repu-
table, well-maintained accommodations that appear clean. Travelers can also re-
duce chances of an infestation at home by washing all clothing and luggage upon
returning home.
Bedbugs can leave itchy bites that typically heal over a few days. The itch common
to the bites can usually be solved with a variety of common bug-bite remedies. A
doctor should be consulted in more serious cases.
Bedbug infestations can be difficult to eliminate, as the pests can hide in a number
of places. Mattresses and carpets suspected to contain bedbugs can be thoroughly
vacuumed to remove the pests, with mattresses then being covered and left in a
sunny place for as long as possible. Bedding and clothing can be washed thoroughly
in hot water. However, professional pest control should also be considered, especially
in larger or repetitive cases.
Anoplura
Sucking lice (Anoplura) have around 500 species and represent the smaller of
the two traditional suborders of lice. The Anoplura are all blood-feeding ectoparasites
of mammals. They can cause localised skin irritations and are vectors of several
blood-borne diseases.
At least three species of Anoplura are parasites of humans. Pediculus humanus is
divided into two subspecies, Pediculus humanus humanus, or the body louse,
sometimes nicknamed «the seam squirrel» for its habit of laying of eggs in the seams
of clothing, and Pediculus humanus capitis, or the head louse. Phthirus pubis (the
pubic louse) is the cause of the embarrassing condition known as crabs.
Head lice
Head lice (Pediculus humanus capitis) are one of the many varieties of sucking
lice (singular «louse») specialized to live on different areas of various animals. As
the name implies, head lice are specialized to live among the hair present on the
human head and are exquisitely adapted to living mainly on the scalp and neck hairs
of their human host. Lice present on other body parts covered by hair are not head
lice but are either Pubic lice (Pthirus pubis) or Body lice (Pediculus humanus
humanus).
Description: The adult head louse resembles a miniature ant that appears flat
when viewed from the side through a strong magnifying glass (Fig. 40). Head lice
have a head, thorax and abdomen with six legs, but their two front legs are very large
in order to grab onto the hair shafts. Head lice are tan to greyish-white in color.
Life cycle: Lice eggs on the hair very close to the scalp are the primary sign of an
active infestation. The female louse glues her eggs, sometimes called «nits», which
look like tiny white beads, to hair shafts very close to the scalp. Eggs are very small,
about the size of a period (full stop) in normal printing. Eggs may appear yellowish,
137
 Species of order Hemiptera occur worldwide; they are distinguished from all
other insects by both adults and nymphs having piercing and sucking mouthparts
housed in a long «beak». These are used mostly to feed on plant juices, but some
species are adapted to suck blood from animals or other insects.
Bedbugs
Bedbugs (or bed bugs) are small nocturnal insects of the family Cimicidae that
live by hematophagy, feeding on the blood of humans and other warm-blooded hosts.
The common bedbug (Cimex lectularius) is the best adapted to human environments.
It is found in temperate climates throughout the world and has been known since
ancient times.
Bedbugs are small, brown, wingless parasites that feed off blood (Fig. 39). Even
though they cannot fly, they are not easily seen, as they are less than a quarter-of-an-
inch (half a centimeter) long and often hide during the day. While bedbugs prefer to
bite humans, they will also feed from other mammals if necessary.
Bedbugs get their name from the fact they often live in unsanitary mattresses and
bedding, but they can be found in other places such as carpets and cracks in walls.
Although they are often found in dirty accommodation where poor sanitation is a
problem, bedbugs have been known to travel in a person’s clothing or luggage to
other locations.
One sign of bedbugs is finding spots of blood in or around beds. Bedbugs typically
leave tiny, itchy bites in orderly rows on a body. Some may notice that rooms that
have many bedbugs have a sweet smell.
Bedbugs are mainly prevented by good sanitation and frequent cleaning, such as
regular housecleaning and washing of bedding. As bedbugs are found worldwide,
travelers abroad should also be watchful for signs of infestation. Travelers in rustic
Figure 39. Bedbug
136
Clinical Disease
Many people infected with T. cruzi remain asymptomatic and free from Chagas’
disease or experience only an acute infection without progressing to the chronic stage.
The most severe form of the disease is most commonly seen in children younger than
5 years of age. Multiplication of T. cruzi at the site of infection can produce an inflamed
swelling (chagoma) which persists for weeks. Trypomastigotes or amastigotes may be
seen in the aspirate of the chagoma. Regional lymph nodes may become infected which
frequently involve one side of the face. Unilateral oedema of the upper and lower
eyelid may occur along with conjuctivitis. This is known as Romana’s sign.
In the acute stage of infection trypomastigotes can be found in the blood. Symptoms
may pass unnoticed, but there may be fever, malaise increased pulse rate and enlargement
of lymph glands, liver and possibly spleen. Muscle aches and pains are characteristic
at this stage and parasites may be seen in blood films. The acute form is most often
seen in young children and occasionally can cause serious damage to the heart and
other complications leading to death caused by central nervous system involvement.
Chronic manifestations include signs of cardiac muscle damage with a weak
and irregular heartbeat, oedema, heart enlargement leading to heart failure. Dilation
of the digestive tract resulting in megaoesophagus and megacolon may also occur.
About 10% of persons infected with T. cruzi develop chronic Chagas cardiopathy.
Laboratory Diagnosis of South American trypanosomiasis
Laboratory diagnosis of South American trypanosomiasis is by
1. Examination of blood.
2. Xenodiagnosis
3. Blood culture
4. Serology
1. Examination of blood
a) Thick and thin blood films are made and stained with Fields stain and examined
as for malaria parasites. Wet preparations of blood can also be examined for motile
trypanosomes.
b) Buffy coat examination – Trypanosomes are centrifuged in a microhaematoctit
tube for 5 minutes. Parasites can be seen microscopically at the junction of the packed
red cells and plasma. This technique is rapid and sensitive.
Trypanosoma cruzi can often be seen in C, U or S shapes in stained films.
2. Xenodiagnosis
Xenodiagnosis is useful in chronic and sub acute (low parasitaemia) disease.
Sterile bugs are fed on patients by attaching a black bag containing the bugs to the
arm of the patient and allowing them to feed for 30 minutes. Twenty five to thirty
days later the bugs are dissected and the contents of the hindgut and rectum are
examined microscopically for the presence of trypanosomes.
29
 3. Blood Culture
Blood culture is as sensitive as xenodiagnosis but it requires sterile conditions.
4. Serology
Serology tests include:
(²) IFAT indirect fluorescence antibody test
(²²) CFT complement fixation test
(²²²) IHAT indirect haemaglutination test
(²V) ELISA enzyme linked immunoabsorbent assay
Other lab findings include:
Raised ESR, marked lymphocytosis with atypical mononuclear lymphocytes
NB. In certain areas of S. America where Trypanosoma rangeli (non pathogenic
species transmitted by Rhodnius bug) is found with T. cruzi all positive preparations
should be checked to confirm T. cruzi.
Leishmania species
Introduction
Leishmaniasis is caused by parasites of the genus Leishmania and is endemic in
many parts of Africa, Asia and South America. It is transmitted by Phlebotomus
species, sandfly.
Leishmania species are mainly parasites of man and other animals, especially dogs
and rodents. They cause diseases collectively known as Leishmaniasis; causing 3 types
of disease i.e. visceral leishmaniasis, cutaneous leishmaniasis and muco-cutaneous
leishmaniasis. These are all debilitating and disfiguring diseases, which occur throughout
the Old and New World. The parasites are unusual in that they live entirely within the
cells of the reticulo-endothelial cells, they have become perfectly adapted as the proteolytic
enzymes which attack other foreign bosies in the blood stream do not destroy them.
Visceral leishmaniasis
Human visceral leishmaniasis (VL), sometimes known as Kala-azar, is caused by
Leishmania donovani complex; L. donovani and L. donovani infantum in the Old World
and L. donovani chagasi in the New World (Fig. 6). The clinical features –azar caused
by these species are similar, but they have different epidemiological features. The parent
species L. donovani occurs in Asia (Northeastern China, India and Iran) and Africa
(primarily Sudan, Kenya and Ethiopia) and can affect people of all ages. The parasite (L.
d. infantum), which causes VL in countries bordering the Mediterranean, (Southern Europe
as well as North Africa) affects young children as well as infants. It is now being seen in
the immunocompromised. In the New World also, VL is mainly a disease of young
children, with the causative organism L. d. chagasi being closely related to, but slightly
30
they eat garbage. They are called the custodians of nature. They only live in houses
where there are crumbs to eat or the garbage can is uncovered. They lay eggs inside
the house’s hollow walls.
The roach is also one of the hardiest insects on the planet, capable of living for a
month without food and remaining alive headless for up to a week. It can also hold
its breath for 45 minutes and has the ability to slow down its heart rate. Cockroaches
also have a very high resistance to radiation.
Select species
! Periplaneta americana, American cockroach
! Eurycotis floridana, Florida woods cockroach
! Blatta orientalis, Oriental cockroach
! Blattella germanica, German cockroach
! Blattella asahinai, Asian cockroach
! Pycnoscelus surinamensis, Surinam cockroach
! Supella longipalpa, Brown-banded cockroach
! Periplaneta australasiae, Australian cockroach
! Periplaneta fuliginosa, Smokybrown cockroach
! Parcoblatta pennsylvanica, Pennsylvania woods cockroach
! Periplaneta brunnea, Brown cockroach
Behavior: New research being conducted at the University of Florida shows
that cockroaches leave chemical trails in their feces. Other cockroaches will
follow these trails to discover sources of food, water, and where other cock-
roaches are hiding. One of the major implications of this research is a new tech-
nique in cockroach pest control. Cockroaches could be potentially removed from
a home by leaving a chemical trail that leads away from the home.
Miscellaneous:
! The largest known cockroach by wingspan is a Megaloblatta longipennis, with
an 18-cm wingspan.
! The largest by weight is a 50g Macropanesthia rhinoceros.
! The smallest species is Attaphila fungicola, reaching only 4 mm.
Heteroptera
Heteroptera (also called true bugs) is a suborder of the order Hemiptera. There
are 25,000 known species in over 60 families. The name Heteroptera comes from
their forewings having both membranous and hard portions. Suborder Heteroptera
includes 25,000 known species in over 60 families.
Hemiptera is an order of insects, comprising some 67,500 known species in two
suborders, Heteroptera and Homoptera. Originally the Homoptera were treated as
a separate order. Members of the Hemiptera, and of the Heteroptera in particular,
are sometimes called «true bugs». The name «heteroptera» comes from their forewings
having both membranous and hard portions. It is also this, which gives the order its
name, hemiptera, coming from the Greek for half-wing.
135
 Human attempts to control pests by insecticides can backfire, because important
but unrecognized insects already helping to control pest populations are also killed
by the poison, leading eventually to population explosions of the pest species.
Cockroaches

Cockroaches are insects of the order Blattodea (the name Blattaria is also seen).
The names of the order are derived from Greek blatta, meaning «cockroach». There
are roughly 3,500 species in 6 families. Cockroaches exist worldwide, with the
exception of the polar regions and in elevations above 2,000 m (6,500 ft).
Among the most well-known species are the American cockroach, Periplaneta
americana, which is about 3 cm long, and the German cockroach, Blattella germanica,
about 1.5 cm long. Tropical cockroaches are often much bigger. When infesting
buildings, cockroaches are considered pests.
The earliest fossils of cockroaches are from the Carboniferous period between
354–295 million years ago.
Biology: Cockroaches are generally either scavengers or omnivores. The
exception to this is the wood eating Cryptocercus species found in China and the
United States. Although they are incapable of digesting the cellulose themselves,
they have a symbiotic relationship with a protozoan that digests the cellulose, allowing
them to extract the nutrients. In this, they are similar to termites. They are most
common in tropical and subtropical climates. Some species are in close association
with human dwellings and widely found around garbage or in the kitchen.
Female cockroaches are sometimes seen carrying egg cases on the end of their
abdomen; the egg case of the German Cockroach holds about 30-40 long, thin eggs,
packed like frankfurters in the case called an ootheca.
The eggs hatch from the combined pressure of the hatchlings gulping air and are
initially bright white nymphs that continue inflating themselves with air and harden
and darken within about four hours. Their transient white stage while hatching and
Figure 6. Leishmania donovani
later while molting has led to many individuals claiming to have seen albino cockroaches.
A female German cockroach carries an egg capsule containing around 40 eggs.
She drops the capsule prior to hatching. Development from eggs to adults takes 3-4 different from, L. donovani. The main geographical foci of VL in Latin America are in
months. Cockroaches live up to a year. The female may produce up to eight egg cases northern and northeastern Brazil. Small foci are found in northern Argentina, Columbia
in a lifetime. In other words, in favorable conditions it can produce 300-400 offspring. and Venezuela. Sporadic cases are found in Central American countries, including Mexico.
A regular cockroach, however, can produce an extremely high number of eggs in
her lifetime. She lays up to 100 eggs in each egg sac. She only needs to be impregnated Cutaneous leishmaniasis
once to be able to lay eggs for the rest of her life, allowing one single cockroach to
lay over a million eggs in her lifetime. Cutaneous leishmaniasis is caused by L. tropica, L. major and L. aethiopica in
The world’s largest cockroach is the Australian giant burrowing cockroach, which the Old World and L. mexicana complex in the New World (Fig. 7). Leishmania
can grow to 9 cm in length and weigh more than 30 grams. Comparable in size is the tropica is widely distributed around the Mediterranean basin, Afghanistan, Kenya,
giant cockroach Blaberus giganteus, which grows to a similar length but is not as massive. Kenya, Armenia, Azerbaijan, Turkmenistan and Uzbekistan. Leishmania aethiopica
Cockroaches are mainly nocturnal, and will run away when exposed to light. A is seen in the highlands of Ethiopia and L. major occurs in the Middle East, West
peculiar exception is the Oriental Cockroach which is attracted to light, thus making Africa, North Africa and Kenya. Leishmania mexicana complex is found in Central
it a far more annoying pest. Roaches are actually very clean insects, even though America and the Amazon Basin.
134 31
 Social insects, such as the ant and the bee, are the most familiar species of eusocial
animal. They live together in large well-organized colonies that are so tightly inte-
grated and genetically similar the colonies are sometimes considered superorganisms.
Roles in the environment and human society: Many insects are considered
pests by humans, because they transmit diseases (mosquitos, flies), damage struc-
tures (termites), or destroy agricultural goods (locusts, weevils). Many entomolo-
gists are involved in various forms of pest control, often using insecticides, but more
and more relying on methods of biocontrol.
Although pest insects attract the most attention, many insects are beneficial to
the environment and to humans. Some pollinate flowering plants (for example wasps,
bees, butterflies, ants). Pollination is a trade between plants, which need to reproduce,
and pollinators which receive rewards of nectar and pollen. A serious environmental
problem today is the decline of populations of pollinator insects, and a number of
species of insects are now cultured primarily for pollination management in order to
have sufficient pollinators in the field, orchard or greenhouse at bloom time.
Insects also produce useful substances such as honey, wax, lacquer or silk.
Honeybees, (pictured above) have been cultured by humans for thousands of years
for honey, although contracting for crop pollination is becoming more significant
for beekeepers. The silkworm has greatly affected human history as silk-driven trade
established relationships between China and the rest of the world. Fly larvae (maggots)
were formerly used to treat wounds to prevent or stop gangrene, as they would only
consume dead flesh. This treatment is finding modern usage in some hospitals. Insect
larvae of various kinds are also commonly used as fishing bait.
In some parts of the world, insects are used for human food («Entomophagy»),
while being a taboo in other places. There are proponents of developing this use to
provide a major source of protein in human nutrition. Since it is impossible to entirely
eliminate pest insects from the human food chain, insects already are present in
many foods, especially grains. Most people do not realize that food laws in many
countries do not prohibit insect parts in food, but rather limit the quantity. According
to cultural materialist anthropologist Marvin Harris, the eating of insects is taboo in
cultures that have protein sources that require less work like farm birds or cattle.
Many insects, especially beetles, are scavengers, feeding on dead animals and
fallen trees, recycling the biological materials into forms found useful by other
organisms. The ancient Egyptian religion adored beetles and represented them as
scarabeums.
Although mostly unnoticed by most humans, arguably the most useful of all insects
are insectivores, those that feed on other insects. Many insects, such as grasshoppers
can potentially reproduce so fast that they could literally bury the earth in a single
season. However there are hundreds of other insect species that feed on grasshopper
eggs, and some that feed on grasshopper adults. This role in ecology is usually assumed
to be primarily one of birds, but insects, though less glamorous, are much more
significant. For any pest insect one can name, there is a species of wasp that is either
Figure 7. Leishmania tropica a parasitoid or predator upon that pest, and plays a significant role in controlling it.
32 133
 Insects have a complete digestive system. That is, their digestive system consists
basically of a tube that runs from mouth to anus, contrasting with the incomplete
digestive systems found in many simpler invertebrates. The excretory system con-
sists of Malpighian tubules for the removal of nitrogenous wastes and the hindgut
for osmoregulation. At the end of the hindgut, insects are able to reabsorb water
along with potassium and sodium ions. Therefore, insects don’t usually excrete wa-
ter with their feces, a fact, which allows them to store water in the body. This process
of reabsorption enables them to withstand hot, dry environments.
Most insects have two pairs of wings located on the second and third thoracic
segments. Insects are the only invertebrate group to have developed flight, and
this has played an important part in their success. The winged insects, and their
secondarily wingless relatives, make up the subclass Pterygota. Insect flight is not
very well understood, relying heavily on turbulent atmospheric effects. In more
primitive insects it tends to rely heavily on direct flight muscles, which act upon
the wing structure. More advanced flyers, which make up the Neoptera, generally
have wings that can be folded over their back, keeping them out of the way when
not in use. In these insects, the wings are powered mainly by indirect flight muscles
that move the wings by stressing the thorax wall. These muscles are able to contract
when stretched without nervous impulses, allowing the wings to beat much faster
than would be otherwise possible.
Insects use tracheal respiration in order to transport oxygen through their bodies.
Openings on the surface of the body called spiracles lead to the tubular tracheal
system. Air reaches internal tissues via this system of branching trachea. The
circulatory system of insects, like that of other arthropods, is open: the heart pumps
the hemolymph through arteries to open spaces surrounding the internal organs;
when the heart relaxes, the hemolymph seeps back into the heart.
Insects hatch from eggs, and undergo a series of moults as they develop and
grow in size. This manner of growth is necessitated by the exoskeleton. Moulting is
a process by which the individual escapes the confines of the exoskeleton in order to
increase in size, then grows a new outer covering. In most types of insects, the young,
called nymphs, are basically similar in form to the adults (an example is the
grasshopper), though wings are not developed until the adult stage. This is called
incomplete metamorphosis. Complete metamorphosis distinguishes the
Endopterygota, which includes many of the most successful insect groups. In these
species, an egg hatches to produce a larva, which is generally worm-like in form.
The larva grows and eventually becomes a pupa, a stage sealed within a cocoon or
chrysalis in some species. In the pupal stage, the insect undergoes considerable change
in form to emerge as an adult, or imago. Butterflies are an example of an insect that
undergoes complete metamorphosis.
Behaviour: Many insects possess very refined organs of perception. In some
cases, their senses can be more capable than humans. For example, bees can see in
the ultraviolet spectrum, and male moths have a specialized sense of smell that enables
them to detect the pheromones of female moths over distances of many kilometers.
132
Mucocutaneous leishmania
Is caused by the L. braziliensis complex and is found in Brazil, Eastern Peru,
Bolivia, Paraguay, Ecuador, Columbia and Venezuela (Fig. 8).
Life cycle
All forms of infection starts when a female sandfly (Phlebotomus species) takes
a blood meal from an infected host. Small amounts of blood, lymph and macrophages
infected with Leishmania amastigotes are ingested. Once ingested the amastigotes
transform to promastigotes in the sandfly, the non-infective promastigotes divide
and develop into infective metacyclic promastigotes. These are formed in the midgut
of the sandfly and migrate to the proboscis. When the sandfly bites the extracellular
inoculated promastigotes at the site of the bite is phagocytosed by macrophages.
After phagocytosis, transformation to dividing amastigotes occurs within 24 hours.
Reproduction at all stages of the lifecycle is believed to occur by binary fission. No
sexual stage has been identified.
Morphology
Leishmania exist as flagellated extracellular promastigotes in the Sandfly vector
and as a flagellar obligate intracellular amastigotes within mononuclear phagocytes
of their vertebrate hosts. The various species are not distinguishable
morphologically from one another. When stained with Romanowsky stains such
as Giemsa, amastigotes appear as round or oval bodies ranging from 2-3mm in
diameter with a well defined nucleus and kinetoplast, a rod shaped specialised
mitochondrial structure that contains extranuclear DNA. The flagellated
promastigote form is spindle shaped, measuring 10-20mm in length, not including
the length of the flagellum. As in the amastigote form a nucleus and kinetoplast are
clearly visible.
Clinical Disease – Visceral leishmaniasis
The incubation period of VL may vary between 2 weeks and 18 months. The
onset of VL is usually insidious with fever, sweating, weakness and weight loss. The
most prominent findings are fever, hepatosplenomegaly and anaemia. The sites mainly
affected are the liver, spleen and bone marrow. Enlargement of the liver is due to
hyperplasia of Kupffer cells which are packed with amastigotes. The bone marrow
is infiltrated with parasitised macrophages. Some organs, notably the kidneys, may
show pathological changes secondary to deposition of immune complexes. In
advanced cases, ascites and oedema can develop. Deaths are usually due to secondary
bacterial infections such as pneumonia, tuberculosis or dysentery.
Laboratory Diagnosis of visceral leishmaniasis
1. Microscopy
Parasites may be found in a splenic aspirate, liver biopsy or bone marrow biopsy.
These techniques, especially splenic aspirate and liver biopsy can be hazardous and
require previous expertise in the procedure.
a) Air dry smears.
b) Fix in methanol for 1 minute
33

Figure 8. Leishmania brasiliens
c) Stain with Giemsa 1 in 10 in buffered distilled water pH 6.8 for 30 minutes (or
use the rapid Field’s stain)
d) Wash the slide in buffered water and drain dry
Amastigotes of leishmania should be seen in positive smears. They are
approximately 2-4µm in size, oval and are frequently seen within the cytoplasm of
the macrophage. The amastigotes possess a nucleus and a rod – shaped kinetoplast
within the cytoplasm. In many samples a very small number of parasites are present.
Extensive searching of the film is necessary.
34
begin to rise. At this time, they resume the quest for hosts in a last-ditch effort to
obtain a blood meal allowing them to mate and reproduce. This second activity peak
typically occurs in March and early April.
Adult female ticks that attach to deer, whether in the fall or spring, feed for
approximately one week. Males feed only intermittently. Mating may take place on
or off the host, and is required for the female’s successful completion of the blood
meal. The females then drop off the host, become gravid, lay their eggs underneath
leaf litter in early spring, and die. Each female lays approximately 3,000 eggs. The
eggs hatch later in the summer, beginning the two-year cycle anew.
TOPIC: INSECTA, DIPTERA
Insects
Insects are invertebrate animals of the Class Insecta, the largest and (on land)
most widely distributed taxon within the Phylum Arthropoda. Insects comprise the
most diverse group of animals on the earth, with over 800,000 species described—
more than all other animal groups combined: «Indeed, in no one of her works has
Nature more fully displayed her exhaustless ingenuity,» Pliny exclaimed. Insects
may be found in nearly all environments on the planet, although only a small number
of species have adapted to life in the oceans where crustaceans tend to predominate.
There are approximately 5,000 dragonfly species, 2,000 praying mantis, 20,000 grass-
hopper, 170,000 butterfly and moth, 120,000 fly, 82,000 true bug, 350,000 beetle,
and 110,000 bee and ant species. Estimates of the total number of current species,
including those not yet known to science, range from two to thirty million, with most
authorities favouring a figure midway between these extremes. The study of insects
is called entomology.
Relationship to other arthropods: A few smaller groups with similar body
plans, such as springtails (Collembola), are united with the insects in the
Subphylum Hexapoda. The true insects (that is, species classified in the Class
Insecta) are distinguished from all other arthropods in part by having
ectognathous, or exposed, mouthparts and eleven (11) abdominal segments. Most
species, but by no means all, have wings as adults. Terrestrial arthropods, such
as centipedes, millipedes, scorpions and spiders, are sometimes confused with
insects due to the fact that both have similar body plans, sharing (as do all ar-
thropods) a jointed exoskeleton.
Morphology and development: Insects range in size from less than a millimeter
to over 18 centimeters (some walkingsticks) in length. Insects possess segmented
bodies supported by an exoskeleton, a hard outer covering made mostly of chitin.
The body is divided into a head, a thorax, and an abdomen. The head supports a pair
of sensory antennae, a pair of compound eyes, and a mouth. The thorax has six legs
(one pair per segment) and wings (if present in the species). The abdomen has
excretory and reproductive structures.
131
 Larva: Eggs laid by an adult female deer tick in the spring hatch into larvae later
in the summer. These larvae reach their peak activity in August. No bigger than a
newsprinted period, a larva will wait on the ground until a small mammal or bird
brushes up against it. The larva then attaches itself to its host, begins feeding, and
engorges with blood over several days.
If the host is already infected with the Lyme disease spirochete from previous
tick bites, the larva will likely become infected as well. In this way, infected hosts in
the wild (primarily white-footed mice, which exist in large numbers in Lyme-endemic
areas of the northeast and upper mid-west) serve as spirochete reservoirs, infecting
ticks that feed upon them. Other mammals and ground-feeding birds may also serve
as reservoirs.
Because deer tick larvae are not born infected, it is believed that they cannot transmit
Lyme disease to their human hosts. Instead, «reservoir» hosts, as mentioned above,
can infect the larvae. Having already fed, an infected larva will not seek another host,
human or otherwise, until after it reaches the next stage in its life cycle. It is not
completely known whether larvae, in themselves, pose a threat to humans or their pets.
Nymph: Most larvae, after feeding, drop off their hosts and molt, or transform,
into nymphs in the fall. The nymphs can remain active throughout the winter and
early spring.
In May, nymphal activity begins. Host-seeking nymphs wait on vegetation near
the ground for a small mammal or bird to approach. The nymph will then latch on to
its host and feed for 4 or 5 days, engorging with blood and swelling to many times its
original size. If previously infected during its larval stage, the nymph may transmit
the Lyme disease spirochete to its host. If not previously infected, the nymph may
become infected if its host carries the Lyme disease spirochete from previous
infectious tick bites. In highly endemic areas of the northeast, at least 25% of nymphs
have been found to harbor the Lyme disease spirochete.
Also humans are the hosts often that come into contact with infected nymphs during
their peak spring and summer activity. Although the nymphs’ preferred hosts are small
mammals and birds, humans and their pets are suitable substitutes. Because nymphs
are about the size of a poppy seed, they often go unnoticed until fully engorged, and
are therefore responsible for the majority of human Lyme disease cases.
Adult: Once engorged, the nymph drops off its host into the leaf litter and molts
into an adult. These adults actively seek new hosts throughout the fall, waiting up to
3 feet above the ground on stalks of grass or leaf tips to latch onto deer (its preferred
host) or other larger mammals (including humans, dogs, cats, horses, and other
domestic animals). Peak activity for adult deer ticks occurs in late October and early
November. Of adults sampled in highly endemic areas of the northeast, at least 50%
have been found to carry the Lyme disease spirochete.
As winter closes in, adult ticks unsuccessful in finding hosts take cover under
leaf litter or other surface vegetation, becoming inactive when covered by ice and
snow. Generally, winters in the northeast and upper mid-west are cold enough to
keep adult ticks at bay until late February or early March but not when temperatures
130
2. Culture
The aspirates can be cultured in Novy-Nicolle-MacNeal (NNN) or Schneider’s
Drosophila medium. In culture the amastigote stage converts to the promastigote
stage. However, this is not a rapid technique, as the parasites may take anything
from 10-21 days to grow.
3. Serodiagnosis
VL produces large amounts of specific IgG, which can be used for diagnosis.
Currently the most used sero diagnostic tests are Indirect-immuno Fluorescent
Antibody Test (IFAT), Enzyme Linked Immunosorbent Assay (ELISA) and Direct
Agglutination Test (DAT).
Clinical Disease – Cutaneous Leishmaniasis
Following a bite from an infected sandfly, a small red papule appears at the site
of the bite about 2-8 weeks later. The papule increases in size centrifugally. The
patient then mounts either a hypersensitive response or an anergic response. In a
hypersensitive response, the papule eventually ulcerates, becomes depressed and
then eventually heals through scarring. The patient is now immune from subsequent
bites. In an anergic response, the nodule grows and spreads over large areas of skin.
This resembles leprosy.
Laboratory Diagnosis of Cutaneous leishmaniasis
1. Slit skin smear.
The margin of the lesion contains amastigotes whereas the centre contains debris
and dead skin material. This margin of the lesion is aseptically punctured with a
hypodermic needle and syringe containing a small amount of saline. The aspirate,
which is drawn up into, the needle is examined microscopically and/or cultured
using the method described in visceral leishmaniasis.
2. Polymerase chain reaction
Gene amplification techniques are powerful and sensitive methods and are useful
in diagnosis of cutaneous leishmaniasis particularly when organisms cannot be
detected microscopically. It is also very useful for the speciation of Leishmania
parasites thus the correct treatment can be administered.
Clinical Disease – Mucocutaneous leishmaniasis
Mucocutaneous leishmaniasis or espundia initially develops like cutaneous
leishmaniasis but develops into lesions in the mucocutaneous junction of the pharynx
resulting in the break down of the palate of the mouth and nose or more rarely the
genitalia or anus. This occurs from a few weeks to several years after the cutaneous
lesion has healed. These lesions result in disfiguring deformities of the nose and
mouth.
Laboratory Diagnosis of Mucocutaneous leishmaniasis
1. Microscopy
Finding the organisms in a histological section of the lesion provides definitive
diagnosis of mucocutaneous leishmaniasis. However, the organisms are very rare in
this form of the disease and culture can be a more sensitive method (see visceral
leishmaniasis).
35
 2. Polymerase chain reaction
The PCR method has the advantage of not only low numbers of parasites in
aspirates but also histological sections. This makes this a very sensitive method in
diagnosing mucocutaineous leishmaniasis when parasites are difficult to detect.
TOPIC: APICOMPLEXA, SPOROZOAE, CILIOPHORA
Toxoplasma gondii
Introduction
Toxoplasma gondii, the causative organism of toxoplasmosis, was first observed in
1927 in the gondi, a North African rodent. The first human case of toxoplasmosis was
also reported that year. The organism is a coccidian protozoa belonging to the subphylum
Apicomplexa and has a world wide distribution occurring in all warm-blooded animals.
Cats are the definitive hosts and they become infected by ingesting oocysts or
cysts in tissues of paratenic hosts, such as mice, or transplacentally. Man becomes
infected either by direct ingestion of oocysts from a cat or by eating raw or
undercooked meat. Those who handle raw meat are particularly at risk. Infection
can be transmitted transplacentally.
Life cycle
The development of the entereoepithelial (sexual) cycle in a cats intestine is
brought about by the ingestion of sporulated oocysts of a mouse with cysts. The pre-
patent period up to the shedding of the oocysts varies with the stage of T. gondii
ingested, for example only 3-10 days if the cat has ingested a mouse containing
cysts, but about 19-20 days or longer after direct infection with oocysts or ingestion
of a mouse containing only tachyzoites. Women most at risk of delivering an infected
infant are those who acquire the infection just prior to gestation (Fig. 9).
Humans can acquire infection by;
1) Accidental ingestion of the oocyst shed in the cats’ faeces
2) Ingestion of the tachyzoite in infected milk or transplacentally
3) Ingestion of the tissue cyst in undercooked or raw meat.
4) Transplant of an infected organ in a seronegative recipient
Clinical Disease
Serological evidence has shown that approximately one third of the world’s
population has Toxoplasma antibodies. This suggests that the majority of infections
are benign with most people exhibiting few or no symptoms, but fever and swelling
may be seen. However Toxoplasma gondii can cause severe illness in congenital
infections, acquired infections and in immunocompromised patients. This may lead
to ocular toxoplasmosis and ultimately to fatal CNS disorders such as encephalitis.
Congenital toxoplasmosis
This occurs approximately in 1 per 1000 pregnancies. It can cause severe damage
to and even death of the foetus. Proliferation of tachyzoites leads to intracellular
calcification, corioretinitis, hydrocephaly, psychomotor disturbances and convulsions.
36
to feed, while hard ticks will attach themselves to the skin of a host for long periods
of time. Most reside in the Northwestern US. Tick bites look like mosquito bites, but
can also sometimes bruise or resemble a bullseye.
Ticks as disease vectors
Hard ticks can transmit human diseases such as relapsing fever, Lyme disease,
Rocky Mountain spotted fever, tularemia, equine encephalitis and several forms of
ehrlichiosis. Additionally, they are responsible for transmitting livestock diseases,
including babesiosis and anaplasmosis. Diseases such as HIV/AIDS and malaria
can be transmitted by soft ticks.
Generally, tick-borne diseases correspond to a specific tick-host combination,
and are limited in their geographical extent.
Location: Ticks are often found in tall grass, where they will rest themselves at
the tip of a blade so as to attach themselves to a passing animal or human. It is a
common misconception that the tick can jump from the plant onto the host. Physical
contact is the only method of transportation for ticks. They will generally drop off of
the animal when full, but this may take several days. Ticks contain a structure in
their mouth area that allows them to anchor themselves firmly in place while sucking
blood. Pulling a tick out forcefully may squeeze contents of the tick back into the
bite and often leaves the mouthpiece behind, which may result in infection. Methods
for removing a tick without it leaving its mouthpiece inside the skin include
anesthetizing the tick with a substance such as ether.
According to the Rhode Island Department of Health, roughly 70% of people
who develop Lyme disease catch it from ticks in their own yard. Some ways to
reduce the risk of this happening include:
! Eliminating salt licks, birdbaths, and other features that attract wildlife;
! Keeping the grass short and free of clippings, leaves, and other debris that
shelter ticks;
! Building a fence to keep out deer, since deer can carry hundreds of ticks;
! Creating a 3-foot wide, 3-inch deep gravel border between the yard and any
adjacent wooded areas.
Life cycle: Deer (black-legged) tick. The deer (or black-legged) tick, and the
related western black-legged tick, are the primary known transmitters of Lyme disease
in the United States. Both are hard-bodied ticks with a two-year life cycle. Like all
species of ticks, deer ticks and their relatives require a blood meal to progress to
each successive stage in their life cycles.
The life cycle of the deer tick comprises three growth stages: the larva, nymph
and adult. In both the northeastern and mid-western U.S., where Lyme disease has
become prevalent, it takes about two years for the tick to hatch from the egg, go
through all three stages, reproduce, and then die. A detailed description of this life
cycle and the seasonal timing of peak activity, as they occur in these regions, is
provided below. The best way to get rid of a tick is to rub it with rubbing alcohol or
hold a flame to it.
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 It is quite easy to look for your own demodex mites, by carefully removing an
eyelash or eyebrow hair and placing it under a microscope. A related species of
demodex mite, (Demodex canis), lives on the domestic dog. While, like with hu-
mans, most dogs live with their mites without harm, a minority does not have im-
mune systems capable of completely controlling the mites, leading to a potentially
dangerous infestation called demodectic mange. While direct treatment for severe
cases is possible using a drug known as Mitaban, which is applied to the animal’s
skin, improved nutrition and checking for other, immune-system suppressing dis-
eases are also recommended.
Demodex mites are also known to cause acne in most teenagers going through
puberty. Because of the excessive sebum production, the mites become more active
and this in turn causes acne. When a large amount of demodex mites are present in a
hair follicle, it can cause the area to become infected and in return leaving pimples
or small bumps on the face.

Ixodid

Ixodid or hard ticks are ticks of the family Ixodidae. Hard ticks are distinguished
from Argasidae or soft ticks by their chitinous external shell. Hard ticks may remain
attached to the skin of a host for long periods of time. Most tick-borne diseases are
carried by hard ticks, although there are some exceptions, such as relapsing fever.
Tick is the common name for the small wingless arachnids that, along with
mites, constitute the order Acarina. Ticks are external parasites, living by
hematophagy on the blood of mammals, birds, and occasionally reptiles and
amphibians. Ticks are important vectors of a number of human and animal
diseases (Fig. 38).
Characteristics: The major families of tick include the Ixodidae or hard ticks,
which have thick outer shells made of chitin, and Argasidae or soft ticks, which have
a membraneous outer surface. Soft ticks typically live in crevices and emerge briefly

Figure 38. Stages of ticks’ development Figure 9. Toxoplasma gondii

128 37
A small, proportion of babies who are asymptomatic at birth develop retinocoroiditis
or mental retardation as children or young adults. When a mother is first exposed to
the parasite in later pregnancy the infant is likely to be less severely damaged or
asymptomatic.
Acquired infections
Infections with T. gondii are often mild with flu-like symptoms thus they often
go unnoticed. However lympadenopathy is the most easily recognized symptom and
it can be accompanied by fever, headache and myalgia. Toxoplasmosis may also
produce infectious mononucleosis like symptoms. Ocular toxoplasmosis is also a
common manifest however it is not yet proven whether this is due to congenital or
acquired infections. Other manifestations of Toxoplasma infections are
meningoencephalitis, hepatitis, pneumonitis and myocarditis.
Immunocompromised patients
Toxoplasmosis has been shown to occur as an opportunistic pathogen in
immunocompromised patients and can cause severe complications. Toxoplasmosis in
immunocompromised patients almost always arises from a reactivation of latent infections.
Conditions, which can predispose to toxoplasmosis are malignancies, organ transplants,
leukaemias and patients with acquired immune deficiency syndrome (AIDS). In
immunocompromised patients, the central nervous system is primarily involved with
diffuse encephalopathy, meningoencephalitis or cerebral mass lesions. Toxoplasma
encephalitis has been reported as a life threatening among patients with AIDS.
Laboratory Diagnosis
1. Serological techniques
The detection of toxoplasma specific antibodies is most commonly used in clinical
laboratories. Specific IgG antibodies typically persist for life whereas specific IgM
antibodies begin to decline after several months. Most laboratories carry out
preliminary tests for IgG antibodies and more definitive tests including IgM and IgA
are carried out in reference laboratories. The Sabin-Feldman Dye Test is the gold
standard for detecting the presence of specific antibodies. It measures the total amount
of specific antibody in a serum, which is capable of participating in antibody-mediated
killing of tachyzoites by complement. This test involves the use of live tachyzoites,
which are derived from infected mice or rats. Because of the use of live organisms,
this test is not recommended in the use of routine laboratories and is thus only
employed in reference centres.
2. Isolation Techniques
Culture of parasites in animals is the best overall method but it can take up to six
weeks before the result is available and is thus a disadvantage. Tissue culture is
more rapid taking three or four days to obtain a result, but is not as sensitive.
3. Antigen detection
The direct detection of very small amounts of specific nucleic acid has been
made possible by the introduction in 1985 of the polymerase chain reaction (PCR).
This technique results in the amplification of a specific fragment of DNA from within
the parasite genome, which is detected by ethidium bromide staining, following gel
38
Treatment
Topical (surface) medications are often effective and must be applied thoroughly
to all skin from the face down, especially to areas known to be primarily affected
(skin folds, etc.). Medication should remain on for more than 12 hours, and prefer-
ably 24, and then washed off. Although the mites may be rapidly killed, improve-
ment is sometimes slow and residual inflammation may take some time to finally
subside. The topical medication of choice is 5% Permethrin because it is safe for all
age groups. Lindane (hexachlorocyclohexane) creams or lotions are considered his-
torical treatments, and should be avoided because they have been shown to have neu-
rotoxic effects in children and infants. Similarly, 5–10% sulfur ointments are consid-
ered historical. A single dose of Ivermectin (dosing: 200 µg/kg) has been reported to
cure, but is an off-label use, and thus considered experimental. Additional topical
treatments include 10% crotamiton (except to eyes, nose, mouth), 25% benzyl ben-
zoate cream or lotion; permethrins offer a simpler, one-application treatment which
may be applied with in a 5% cream that remains on overnight or for 8-14 hours.
A person can be reinfected with scabies. Without a host, scabies mites survive
for a few days. Therefore it is recommended, after treatment, to wash all material
(such as clothes and bedding) that has been in prolonged contact with the infested in
the last four days.
To prevent reinfestation, all social contacts and members of the family, even if
not infested, should be treated similarly and most importantly at the same time.
Approximately 300 million cases of infestation with scabies occur worldwide
annually. Scabies also occurs in dogs; see Mange. Note: Although the mange mites
are not able to complete their life cycle on humans, they will cause quite a bit of
itching before they finally die.
The demodex mite
The demodex mite is a tiny parasitic mite, which lives around human hair follicles,
particularly those of the eyelashes and eyebrows (Demodex folliculorum hominis) or
in sebaceous glands connected to hair follicles (Demodex brevis). Measuring between
0.1mm and 0.4mm, each mite has eight segmented legs for locomotion, a long, scale-
covered body for anchoring itself in the hair follicle, and pin-like mouth-parts for
eating skin-cells and oils which accumulate in the hair follicles. Interestingly, the mite’s
digestive system is so efficient and results in so little waste that there is no excretory
orifice! With a life cycle lasting around two weeks, the mites are transferred between
hosts through contact of hair, eyebrows and of the sebaceous glands on the nose.
A surprising fact is that an estimated 96-98% of all people carry such mites – with
up to 25 in each follicle, each person can have a potentially huge population of mites.
In the vast majority of cases, the mites go unobserved, without any adverse symptoms,
but in certain cases (usually related to a suppressed immune system, caused by stress
or illness) mite populations can dramatically increase, resulting in a condition known
as demodicosis, characterised by itching, inflammation and other skin disorders.
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Figure 37. Sarcoptes scabiei. 1. Epiderm. 2,4. Sarcoptes scabiei.
3. Egg.
burrow. Most burrows occur in the webs of fingers, flexing surfaces of the wrists,
around elbows and armpits, the areolae of the breasts in females and on genitals of
males, along the belt line, and on the lower buttocks. The face usually does not
become involved in adults. The rash may become secondarily infected; scratching the
rash may break the skin and make secondary infection more likely. In persons with
severely reduced immunity, such as those with HIV infection, or people being treated
with immunosuppressive drugs like steroids, a widespread rash with thick scaling may
result. This variety of scabies is called Norwegian scabies. Scabies is frequently
misdiagnosed as intense pruritis (itching of healthy skin) before papular eruptions
form. Upon initial pruritus the burrows appear as small, barely noticeable bumps on
the hands and may be slightly shiny and dark in color rather than red. Initially the
itching may not exactly correlate to the location of these bumps. As the infestation
progresses, these bumps become more red in color. Generally diagnosis is made by
finding burrows, which often may be difficult because they are scarce, because they
are obscured by scratch marks, or by secondary dermatitis (unrelated skin irritation). If
burrows are not found in the primary areas known to be affected, the entire skin sur-
face of the body should be examined.
The suspicious area can be rubbed with ink from a fountain pen or alternately a
topical tetracycline solution, which will glow under a special light. The surface is then
wiped off with an alcohol pad; if the person is infected with scabies, the characteristic
zigzag or S pattern of the burrow across the skin will appear. When a suspected burrow
is found, diagnosis may be confirmed by microscopy of surface scrapings, which are
placed on a slide in glycerol, mineral oil or immersion in oil and covered with a coverslip.
Avoiding potassium hydroxide is necessary because it may dissolve fecal pellets.
Positive diagnosis is made when the mite, ova, or fecal pellets are found.
126
electrophoresis. PCR is so sensitive it should detect Toxoplasma DNA in one cyst.
However this may indicate a latent infection rather than an active infection. However
its sensitivity may create problems since it will detect very small amounts of DNA
from latent as well as active infections and it does not differentiate between cyst and
tachyzoite DNA. Thus samples like blood, CSF, urine and and amniotic fluid should
be used as they do not contain the latent stages. PCR shows great promise but as yet
is still labour intensive and expensive for routine use in the laboratory.
Malaria
Blood Parasites
Red Blood cells offer parasites an excellent environment for invasion and survival.
Haemosporina are the only protozoan parasites, which can invade the red blood
corpuscles of vertebrates. Most, if not all, have multiplicative phases in the reticulo-
endothelial system.
The red blood cells are thin-walled and constantly moving, with the result that
absorption of food materials and elimination of waste products of metabolism are relatively
easy to achieve. In addition, they also contain rich supplies of protein and oxygen.
It is now known that malarial parasites do not actually penetrate the red blood
cell but, in fact, enter the cell membrane by endocytosis and enclosed in a
parasitophorous membrane.
Malaria
Introduction
Malaria is the most important tropical disease known to mankind, causing many
deaths and much morbidity throughout the world. It remains a significant problem in
many tropical areas especially in sub-Saharan Africa. In many areas of the world the
situations deteriorating as a result of environmental changes, including global
warming, civil disturbances, increasing travel and drug resistance. (Greenwood, B.M,
1997) There are approximately 100 million cases of malaria worldwide with about 1
million of these proving fatal.
Malaria is caused by protozoa of the Plasmodium species. There are 4 species
which infect both humans and aniamls; Plasmodium malariae (quartian malaria),
Plasmodium vivax (benign tertian malaria), Plasmodium falciparum (malignant
tertian malaria, subtertian malaria) and Plasmodium ovale (ovale tertian malaria).
The transmission of the protozoa, Plasmodium requires two hosts, an intermediate
invertebrate host (vector), and a definitive vertebrate host (mammals, birds and
lizards).
All Plasmodium species undergo the general haemosporina developmental cycle,
which can be summarised as:
1. Initial or continual schizogny in the vertebrate host with initiation of
gametogeny;
39
 2. Formation of gametes in the arthropod host and subsequent fertilisation and
formation of a zygote;
3. Formation of sporozoites from the zygote by repeated nuclear division followed
by cytoplasmic divisions. (Smyth, J.D, 1994)
There is no requirement for resistant stages since the transfer of the parasites
between the vertebrate and invertebrate hosts is made by withdrawal or injection
during the bloodsucking act, there is little or no exposure to the hazards of the outside
world; thus by blood transfusion or inoculation, via the blood stages of the parasite.
Malaria is transmitted by the female anopheline mosquito. The life cycle of all
species of human malaria parasites is essentially the same (Fig. 10). It comprises an
exogenous sexual phase (sporogony) with multiplication in certain Anopheles
mosquitoes and an endogenous asexual phase (schizogony) with multiplication in
the vertebrate host. The latter phase includes the development cycle in the red cells
(erythrocytic schizogony) and the phase taking place in the parenchyma cells in the
liver (pre-erythrocytic schizogony).
When a female Anopheles mosquito bites an infected person, it ingests blood,
which may contain the mature sexual cells (male and female gametocytes), which
undergo a series of developmental stages in the stomach of the mosquito.
Exflagellation occurs resulting in the production in a number of male and female
gametes. Fertilisation occurs producing a zygote, which matures to an ookinete.
This penetrates the stomach wall of the mosquito where it grows into an oocyst and
it further matures to become a motile sporozoite.
The length of the developmental stage in the mosquito not only depends on the
Plasmodium species but also the mosquito host and the ambient temperature. This may
range from 8 days in Plasmodium vivax to as long as 30 days in Plasmodium malariae.
The sporozoites migrate from the body cavity of the mosquito to the salivary
glands and the mosquito now becomes infective. Sporozoites enter into the blood
stream of a host when the mosquito feeds on blood. Following the inoculation, the
sporozoites leave the blood within 40 minutes and enter the parenchymal cells of the
liver (hepatocytes). In all 4 species, asexual development occurs in the liver cells, a
process known as pre-erythrocytic schizogony, to produce thousands of tiny
merozoites, which are relaeased into the circulation after about 16 days. However in
P. vivax and P. ovale some sporozoites differentiate into hypnozoites, which remain
dormant in hepatocytes for considerable periods of time. When they are “reactivated”
they undergo asexual division and produce a clinical relapse.
In P. falciparum and P. malariae hypnozoites are not formed and the parasite
develops directly into pre-erythrocytic schizonts.
Once in the circulation, the merozoites invade the red cells and develop in to
trophozoites. In the course of their development they absorb the haemoglobin of
the red cells leaving as the product of digestion a pigment called haemozoin, a
combination of haematin and protein. This iron-containing pigment is seen in the
body of the parasite in the form of dark granules, which are more obvious in the later
stages of development.
40
Figure 36. Mites
members of the Sarcoptic Mange mites (family Sarcoptidae), which burrow under the
skin. Perhaps the most well known, though, is the house dust mite (family Pyroglyphidae).
Insects may also have parasitic mites. Examples are Varroa destructor, which
attaches to the body of the honeybee and Acarapis woodi, which lives in the tra-
cheae of honeybees.
The scientific discipline devoted to the study of ticks and mites is called
Acarology.
Sarcoptes scabiei
Etiology
Caused by the mite Sarcoptes scabiei, variety hominis, it produces intense, itchy
skin rashes when the impregnated female tunnels into the stratum corneum of the
skin and deposits eggs in the burrow (Fig. 37). The larvae, which hatch in 3-10 days,
move about on the skin, molt into a «nymphal» stage, and then mature into adult
mites. The adult mites live 3-4 weeks in the host’s skin. The motion of the mite in
and on the skin produces an intense itch, which may resemble an allergic reaction in
appearance. The presence of the eggs produces a massive allergic response, which,
in turn, produces more itching. Scabies is transmitted readily, often throughout an
entire household, by prolonged skin-to-skin contact with an infected person (e.g.
bed partners), and thus is sometimes classed as a sexually transmitted disease. Spread
by clothing, bedding or towels is a less significant risk, though possible.
Signs, Symptoms, and Diagnosis: A delayed hypersensitivity (allergic) response
resulting in a papular eruption (red, elevated area on skin) often occurs 30-40 days
after there may be hundreds of papules, less than 10 burrows are typically found.
The burrow appears as a fine, wavy and slightly scaly line a few millimeters to one
centimeter long. A tiny mite (0.3 to 0.4 mm) may sometimes be seen at the end of the
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are exchanged correctly, the male spider must make a timely departure after mating
to escape before the female’s normal predatory instincts come back into operation.
Unusually, sperm transmission is an indirect process. When a male is ready to mate,
he will spin a web pad onto which the contents of the abdominal reproductive organs
are discharged. He then dips his palps (also known as “palpi”), the small, leg-like
appendages on the front of his cephalothorax, into the sperm, absorbing it. Mature
male spiders characteristically have swollen bulbs on the end of their palps for this
purpose, and this is a useful way to identify the sex of a spider in the field. With his
palps thus “charged” he then goes off in search of a female. The act of copulation
occurs when the male inserts one or both palps into the female’s genital opening,
known as the epigyne. He transfers his sperm into the female by contracting his
palps. Very unusual behaviour is seen in spiders of the genus Tidarren, as the male
amputates one of his palps before maturation and enters his adult life with one palp
only. The palpi constitute 20% of its body mass, and since this weight greatly impedes
its movement, the spider detaches one of the two to gain mobility. In the Yemeni
species Tidarren argo, the remaining palp is then torn off by the female. The separated
palp remains attached to the female’s epigynum for about four hours and apparently
continues to function independently. In the meantime the female feeds on the palpless
male. (Journal of Zoology (2001).
Ecology: Spiders have a great range of variation and lifestyle, although all are
predatory.
While spiders are generalist predators, in actuality their different methods of
prey capture often limits the type of prey taken. Thus web-building spiders rarely
capture caterpillars and crab spiders that ambush prey in flowers capture more bees,
butterflies and some flies than other insects. Groups of families that tend to take
certain types of prey because of their prey capture methods are often called guilds. A
few spiders are more specialized in their prey capture. Dysdera captures and eats
sowbugs, pillbugs and beetles, while pirate spiders eat only other spiders. Bolas
spiders in the family Araneidae use sex pheromone analogs to capture only the males
of certain moth species. Despite their generally broad prey ranges spiders are one of
the most important links in the regulation of the populations of insects. Every day on
a meadow they devour over 10 g/m² of insects and other arthropods. Figure 10. Life cycle of malarial parasites

Mites and ticks
The mites and ticks, order Acarina or Acari, belong to the Arachnida and are
among the most diverse and successful of all the invertebrate groups, although some
way behind the insects (Fig. 36). They have exploited an incredible array of habitats
and because of their small size (some are microscopic) most go totally unnoticed.
Many live freely in the soil or water, but there is also a vast array of species that live
as parasites on plants or animals.
Some of the plant pests include the so-called Spider mites (family Tetranychidae)
and the Gall mites (family Eriophyidae). Among the species that attack animals there are
124
After a period of growth the trophozoite undergoes an asexual division,
erythrocytic schizogony. When the mature trophozoite starts to divide in the red
blood cell, separate merozoites are formed resulting in a schizont. When fully
developed, the schizont ruptures the red blood cell containing it, liberating the
merozoites into the circulation. These merozoites will then infect new red cells and
the process of asexual reproduction in the blood tends to proceed. Some of the
merozoites entering red blood cells do not form trophozoites then schizonts but
develop into gametocytes and this process takes place in deep tissue capillaries.
This erythrocytic cycle of schizogony is repeated over and over again in the course
of infection, leading to a progressive increase of parasitaemia.
41
 Infection with all four strains of malaria has many clinical features in common.
These are related to the liberation of fever-producing substances, especially during
schizogony. The common features are:
Fever: Often irregular. The regular pattern of fever does not occur until the illness
has continued for a week or more. Where it depends on synchronised schizogony.
Anaemia: The anaemia is haemolytic in type. It is more severe in infections with
P. falciparum because in this infection cells of all ages can be invaded. Also, the
parasitaemia in this infection can be much higher than in other malarias.
Splenomegaly: The spleen enlarges early in the acute attack of malaria. When a
patient has been subjected to many attacks, the spleen may be of an enormous size
and lead to secondary hypersplenism.
Jaundice: A mild jaundice due to haemolysis may occur in malaria. Severe
jaundice only occurs in P. falciparum infection, and is due to liver involvement.
Species Specific Characteristics
Plasmodium falciparum
Introduction
Plasmodium falciparum is the most important malaria parasite, found in the tropics
and sub-tropics, being responsible for approximately 50% of all malaria cases. The
incubation period of P. falciaprum malaria is the shortest, between 8 and 11 days
and has a periodicity of 36-48 hours. It can be differentiated from the other species
by the morphology of the different stages found in the peripheral blood. In infections
with Plasmodium falciparum usually only young trophozoites and gametocytes
are seen in peripheral blood smears, the schizonts are usually found in capillaries
sinuses of internal organs and in the bone marrow. The disease it produces runs an
acute course and often terminating fatally. It is a significant cause of abortions and
stillborns and even death of non-immune pregnant women.
Life cycle
The aspects of the life cycle, which are specific to P. falciparum, are as follows:
a) It attacks all ages of erythrocytes so that a high density of parasites can be
reached quickly. In extreme cases up to 48% of the red blood cells can be parasitised.
b) Multiple infections resulting in several ring forms in a corpuscle are not
uncommon.
c) The latter stages in the asexual cycle do not occur in the peripheral blood as
in other forms of malaria, so that only rings and crescents are found in blood films.
After 24 hours the ring forms and older trophozoites show a tendency to clump
together and adhere to the visceral capillary walls and become caught up in the
vessels of the heart, intestine, brain or bone marrow in which the later sexual stages
are completed.
d) Sporulation is not as well synchronised as in other malaria forms so that the
fever may last longer.
42
filled with haemolymph, which is pumped through arteries by a heart into spaces
called sinuses surrounding their organs.
Spiders have developed several different respiratory anatomies, based either on
book lungs, a tracheal system, or both. Primitive mygalomorph spiders generally
have only a pair of book lungs filled with haemolymph, where openings on the
ventral surface of the abdomen allow air to enter and diffuse oxygen. Modern
araneomorph spiders often have a single book lung in addition to spiracles, which
deliver air into the tracheae, where oxygen is then diffused into the haemolymph. In
the tracheal system oxygen interchange is much more efficient, enabling cursorial
hunting (hunting involving rapid pursuit) and other advanced characteristics.
Vision: Spiders usually have eight eyes in various arrangements, a fact which is
used to taxonomically classifiy different species. Sometimes one pair of eyes is better
developed than the rest, or there are only six pair, or no eyes at all. Several families
of hunting spiders have developed well to excellent vision, such as wolf spiders and
jumping spiders. However most spiders that lurk on flowers, webs and other fixed
locations waiting for prey have very poor eyesight, but possess extreme sensitivity
to vibrations for hunting.
Defense: Some primitive spiders, like the tarantula, have a patch of urticating
hairs on their abdomens for defense, which are generally absent on modern spiders.
Certain other species have specialized defense tactics. For example, the Golden
Wheeling spider of the desert escapes Tarantula Wasps (a species of wasp that lays
it’s eggs in a paralyzed spider so the larvae have enough food when they hatch) by
flipping onto its side and cartwheeling away.
Life cycle: The spider life cycle progresses through three stages: the embryonic,
the larval, and the nympho-imaginal. Between the time an egg is fertilized and the
spider begins to take the shape of a spider is referred to as the embryonic stage
(Foelix, 1996). As the spider begins to look more like a spider it enters the larval
stage (Foelix, 1996). It enters the larval stage as a prelarva and, through subsequent
molts; it reaches its larval form, a spider-looking, non self-sufficient animal feeding
off its yolk supply (Foelix, 1996). After a few more molts, also called instars, body
structures become differentiated; all organ systems are complete and the animal
begins to hunt on its own; it has reached the nympho-imaginal stage (Foelix, 1996).
This stage is differentiated by two sub-stages: the nymph, or juvenile stage and the
imago, or adult stage (Foelix, 1996). A spider does not transition from the nymph to
the imago until it has become sexually mature (Foelix, 1996). Once a spider has
reached the imago stage, it will remain there until its death. Many spiders may live
only about a year, but a number will live two years or more, overwintering in sheltered
areas (the annual influx of “outdoor” spiders into houses in the fall is due to this
search for a nice warm place to spend the winter).
Reproduction: Spiders reproduce by eggs laid in silk bundles called egg sacs.
Spiders often use elaborate mating rituals (especially in the visually advanced
jumping spiders) to allow the male to approach close enough to inseminate the fe-
male without triggering a predatory response. Assuming that the approach signals
123

Figure 35. Lathrodectus tredecimguttatus. 1. Pedipalps. 2. Duct of poisoning gland. 3.
Poisoning gland. 4. Eyes. 5. Main nervous node. 6. Stomach. 7. Sucking stomach. 8.
Heart. 9. Liver. 10. Medium intestine. 11. Excretory system. 12. Anal foramen. 13.
Arachnidium. 14. Trachea. 15. Ovary. 16. Genital foramen. 17. Book lungs. 18. Chelicerae.
prosoma) and an abdomen, supported by a hard exoskeleton composed mainly of
chitin (Fig. 35).
Spiders also have eight legs (insects have six), no antennae, and their eyes are
single lenses rather than compound eyes. Additionally spiders have pedipalps (or
just palps), which are two appendages next to their mouths that aid in manipulating
food and are used by the males in mating.
Respiration and circulation: Spiders have an open circulatory system, mean-
ing they don’t have true blood or veins for it to travel in. Rather, their bodies are
122
e) Exo-erythrocytic forms do not persist in the tissues and hence relapses do not occur.
Morphology of trophozoites
Red blood cells in Plasmodium falciparum infections are not enlarged and they
may have a spiky outline, which is common in cells, which have dried slowly. The
typical arrangement cytoplasm in young trophozoites is the well-known ring
formation, which thickens and invariably contains several vacuoles as the trophozoite
develops. Chromatin is characteristically found as a single bead, but double beads
and small curved rod forms frequently occur.
Maurer’s dots are slow to appear and are first seen as minute purplish dots, 6 or
less in number. The points become spots, still few in number and are now characteristic
enough to be recognised. Maurer describes them as fine ringlets, loops or streaks.
They are seldom absent from the red blood cells containing large rings but the staining
of the spots is very sensitive to pH and is seldom seen if the pH falls below 6.8.
Trophozoites of P. falciparum can be found on the edge of the red blood cells.
These are known as acole forms and are found as three distinct types:
1. Common: The single chromatin bead lies on the edge of the cell with most of
the cytoplasm extended along the edge on both sides of the bead.
2. Rim: The complete parasite lies in a thickened line along the edge of the cell
with no evidence of ring formation.
3. Dispalced: The parasites are displaced beyond the edge of the host cell. All
degrees of displacement may occur, from partial to marked displacement with most
of the parasite lying beyond the cell margin.
Gametocytes
Gametocytes are the sexual stage of the malaria parasite. Plasmodium falciparum
gametocytes appear in the peripheral circulation after 8-11 days of patent parasitaemia
and by then, they have assumed their typical crescentic shapes. They soon reach their
peak density, and then decline in numbers, disappearing in about 3 months as a rule.
The female form, or macrogametocyte, is usually more slender and somewhat
longer than the male, and the cytoplasm takes up a deeper blue colour with Giemsa
stain. The nucleus is small and compact, staining dark red, while the pigment granules
are closely aggregated around it. The male form, or microgametocyte, is broader
than the female and is more inclined to be sausage shaped. The cytoplasm is either
pale blue or tinted with pink and the nucleus, which stains dark pink, is large and less
compact than in the female, while the pigment granules are scattered in the cytoplasm
around it.
In humans, gametocytes do not multiply, nor cause symptoms but they are the
forms, which are infective to the mosquito. When a female Anopheline mosquito
takes a blood meal, the male and female gametocytes continue their sexual
development.
Schizonts
Schizonts are rarely seen in the peripheral blood and their presence may indicate
a potentially serious parasitaemia. Schizonts have 8-36 merozoites and a large mass
of golden brown pigment (haemozoin) is seen in the pre-schizont and schizont stage.
43
 Clinical Disease
Symptoms include headache, photophobia, muscle aches and pains, anorexia, nausea
and vomiting. Complications include severe anaemia cerebral malaria, renal disease,
black water fever, dysentery, pulmonary oedema and tropical splenomegaly syndrome.
Plasmodium vivax
Introduction
Plasmodium vivax is found almost everywhere malaria is endemic and is the
most predominant of the malaria parasites. Causing 43% of all cases of malaria in
the world, it also has the widest geographical distribution. Although the disease
itself is not usually life threatening, it can cause severe acute illness.
Plasmodium vivax does not infect West Africans due to the fact that West Africans
do not possess the Duffy Antigen on the red blood cells, which the parasite requires
to enter the red blood cell. It has an incubation period of between 10 and 17 days,
which is sometimes prolonged to months or years due to the formation of hypnozoites.
It has a periodicity of 48 hours. Plasmodium vivax infections are usually characterised
by the presence of more than one developmental stage in the peripheral blood film.
The parasites parasitise young enlarged erythrocytes and Schüffner’s dots develop
on the erythrocyte membrane (Fig.11).
Life cycle
The aspects of the life cycle, which are specific to P. vivax, are as follows:
a) The degree of infectivity is low, only the young immature corpuscles are
infected; about 2% of erythrocytes are parasitised.
b) The periodicity of the asexual cycle is closely synchronised.
c) Hypnozoites develop in the liver, so that relapses may occur.
Morphology of trophozoites
Most trophozoites of P. vivax are already several hours old when they appear in
peripheral blood and by that time the Schüffner’s dots are already visible. The
trophozoites are actively amoeboid and contain single or sometimes double chromatin
dots that are either circular or ovoid. As the trophozoites mature, the Schüffner’s
dots increase in number and size and the parasite changes from large irregular rings
to rounded or ovoid forms in mature trophozoites.
Gametocytes
Mature female gametocytes are large rounded parasites, which fill or nearly fill the
host cell. The cytoplasm is blue and fairly homogenous. The nuclear chromatin is a
single, well-defined purplish mass, varied in form and usually peripheral in distribution.
Mature male gametocytes can be distinguished from females by the large, loose and
ill-defined mass of chromatin and by their paler colour and smaller mass.
Schizonts
The parasitised red cells are much enlarged containing Schüffner’s dots. The
parasites are large, filling the enlarged red cell. There are between 12-24 merozoites
in the schizonts (usually16). The pigment is a golden brown central loose mass.
44
passed to the formidable chelicerae for crushing. Only liquid and very fine particles
are ingested; the pulp that results after a meal is discarded.
Mating takes place when the male encounters a receptive female. No spermato-
phore is produced by the male; instead a sperm droplet is transferred from the substrate
to the female gonopore (genital opening) by means of the male’s chelicerae. In one
southwestern species, the male turns the female on her back, emits seminal fluid
directly into the female gonopore, then tamps it in with his chelicerae.
After mating, the female constructs a burrow and nest in the ground, where she
lays 50-200 eggs. She remains with the eggs until they hatch. When the young
solpugids hatch, they emerge from the burrow with their mother and remain together
for some time while the mother captures prey to feed the entire family.
Envenomation: Solpugids are commonly considered to be venomous, but poison
glands have not been found to be associated with the chelicerae. It has been suggested
(but not confirmed) that poisoning might result from toxins secreted through the
setal (bristle) pores that can be traced along the tips of the chelicerae. Apparently
authentic cases of aftereffects resulting from a solpugid bite have been recorded, but
these symptoms were probably caused by bacterial infection of the wound.
The solpugid has undoubtedly been maligned because of its appearance. An old
wives’ tale holds that animals drinking from a water trough in which a solpugid is
present will die. There is no foundation for this.
Treatment: If a bite results in broken skin, wash with soap and water and apply
antiseptic to the wound.
Precautions and Control: Don’t handle solpugids. If one is found in the house,
it can be brushed into a dustpan or other container and returned to the outdoors.
Solpugids are usual and beneficial components of Arizona’s varied ecosystems. These
animals cannot be controlled, nor would it be desirable to control them.
Spiders
Spiders are invertebrate animals that produce silk, have eight legs and no wings.
More precisely, a spider is any member of the arachnid order Araneae, an order
divided into three sub-orders in newer systems: the Mygalomorphae (the primitive
spiders), the Araneomorphae (the modern spiders) and the Mesothelae, which contains
the Family Liphistiidae, rarely seen burrowing spiders from Asia. The study of spiders
is known as arachnology, although it is often included in the more general term
entomology.
Many spiders hunt by building webs to trap insects. These webs are made of
spider silk, a thin, strong protein strand extruded by the spider from spinnerets on
the end of the abdomen. All spiders produce silk, although not all use it to spin
elaborate traps. Silk can be used to aid in climbing, forming smooth walls for burrows,
cocooning prey, and for many other applications.
Morphology and development: Spiders, unlike insects, have only two body
segments instead of three; a fused head and thorax (called a cephalothorax or
121
the Devonian and the Oligocene; there is a severe lack of fossils known from the
intervening period.
These earliest scorpiones are considered to be Protoscorpions, since they pos-
sess many traits, which are plesiomorphic for scorpions. For example, in all scorpi-
ons the thick front portion on the abdomen is made up of seven segments, but the
number of sternite plates, which cover this region varies among the earliest fossils,
while all living species have five. All scorpions have an additional five segments
after the initial seven, ending in a sharp sting. This sting contains a pair of poison
glands, which can paralyze prey, usually insects or small rodents, or may deliver a
painful sting to incautious persons. Most scorpion stings are merely painful, leading
to swelling in the immediate region of the sting, but some scorpions of northern
Africa and the American southwest can be deadly. In the US, the deadliest scorpions
are to be found in Arizona, where it is a good idea to shake out shoes before putting
them on in the morning! Besides their unusually long and dangerous tails, scorpions
also differ from other arachnids in having large pedipalps. These are the second pair
of appendages on the body, and are usually rather inconspicuous in arachnids, but in
scorpions, they are large and powerful pincers, which may be used to grasp and
subdue prey. Scorpions may also have more eyes than other arachnids, some species
possessing as many as six pairs, though most do not have this many. They have three
joints in their chelicerae, or the first pair of appendages, located next to the mouth.
Most scorpions are nocturnal, hiding under rocks, in crevices, or within burrows
during the day, and coming out after sunset. Because of this, and because of their
painful stings, it can be dangerous to travel at night in scorpion territory without
shoes, even inside homes. (I have nearly stepped on a scorpion in my parents’ home
near Dallas.) One unusual feature of scorpions that has helped many field biologists
is the UV fluorescence of scorpion bodies. Biologists hunting for scorpions wave an
ultraviolet light near the ground as they walk along, watching for an eerie greenish
light to be reflected back. The UV light is absorbed by the scorpion’s armor and is
reflected back as visible light.

Solpugids

Description: The solpugids (sun spiders, wind scorpions; order Solpugida) are
spiderlike and hairy, with two closely placed median eyes. Their most striking
characteristic is the enormous size of their chelicerae. These projects in front of the
head, and each of the pair is composed of two pieces forming a pincer that works in
a vertical plane. The pedipalps are leglike, but have a specialized adhesive terminal
segment. The pedipalps and the first pair of legs (that have a tactile function) are Figure 11. Plasmodium vivax
usually carried above the cephalothorax while the solpugid is standing or running.
Solpugids are common in the hot desert regions of the world. Clinical Disease
Biology: Solpugids are voracious predators that feed on all types of small Symptoms include headache, photophobia, muscle aches and pains, anorexia,
arthropods. While they are most active at night, it is not unusual to see them out and nausea and vomiting. Complications due to P. vivax are relatively rare and arise due
about during the day. Prey is captured with the sticky ends of the pedipalps and do a previous debility or pre-existing disease.
120 45
 Plasmodium ovale
Introduction
Plasmodium ovale is widely distributed in tropical Africa especially the west
coast, despite that it is a species that is rarely encountered. It has also been reported
in South America and Asia. It has an incubation period of 10-17 days, which is
sometimes prolonged to months or years due to the formation of hypnozoites. It
has a periodicity of 48 hours; the fever it produces is milder than the benign tertian
P. falciparum.
Life cycle
The features of the life cycle, which are specific to P. ovale are as follows:
a) It morphologically resembles P. malariae in most of its stages.
b) The changes produced in the erythrocytes in general are similar to those
produced by P. vivax, but Schüffner’s dots appear considerably earlier (in the ring
stage) and are coarser and more numerous.
c) In the oocyst the pigment granules are (usually) characteristically arranged in
two rows crossing each other at right angles.
d) Hypnozoites develop in the liver so that relapses may occur.
Morphology
Parasites of P. ovale are usually found in enlarged and stippled red blood cells
(James’s dots), similar to those found in P. vivax infections. Host cells show an oval
shape, particularly those containing younger stages of the parasites and the host cell
may also show “spiking” or fimbriation.
Trophozoites
Young trophozoites are found as compact rings in cells containing Schüffner’s
dots. The trophozoite rings remain compact as they develop and show little of the
amoeboid features common in P. vivax. Small, scattered pigment granules can be
seen in developing trophozoites, which disperse as the trophozoite matures. Late
trophozoites are round and consolidated with an increase in cytoplasm, they are
very similar to P. vivax at this stage.
Gametocytes
The mature gametocytes are round, filling two thirds of the red cell. The red
blood cell is slightly enlarged and stippled and contains pigment, which has a distinct
arrangement of concentric rods, mostly at the periphery.
Schizonts
The parasite is smaller than red blood cells and contains 6-12 merozoites, usually
8 in a single ring. The pigment is a brown / greenish central clump. The red cell
slightly enlarged, stippled, frequently oval and fimbriated.
Clinical Disease
Symptoms, like those of P. vivax, include headache, photophobia, muscle
aches and pains, anorexia, nausea and vomiting. Complications due to
P. ovale are relatively rare and arise due do a previous debility or pre-existing
disease.
46
birds, and mammals combined! Although the insects still rule in terms of numbers, the
crustaceans are the most diverse in terms of form. The largest of the crustaceans include
the the giant Japanese spider crab (Macrocheira kaempferi) with its four-meter legspan,
the Alaskan king crab (Paralithodes camtschatica), which can weigh more than 10
kilograms, and the giant Tasmanian crab (Pseudocarcinus gigas), which has been recorded
at an impressive 14 kilograms. On the other end of the spectrum, some crustaceans never
grow larger than 0.25 millimeters, even as adults. Crabs, shrimps, and lobsters are well-
known crustaceans. However, barnacles, pillbugs, amphipods, copepods, krill, crayfishes,
sea fleas, clam shrimps, fairy shrimps, and many others also belong to the Crustacea, an
ancient group that arose in the early Cambrian nearly 600 million years ago.
The Arachnida
The Arachnida include the terrestrial chelicerates that everyone is familiar with,
and that nearly everyone would rather not be too familiar with: spiders (Araneae),
ticks and mites (Acari), and scorpions (Scorpiones). Arachnids also include a number
of less familiar taxa: Opiliones (harvestmen or daddy-longlegs); Thelyphonida
(whip-scorpions); Pseudoscorpiones (false scorpions); and many others. Most are
predators, and some are venomous. All are terrestrial, except for some mites and
spiders that have become secondarily aquatic.
Scorpions
Scorpions are the oldest arachnids for which fossils are known, and they were
the first arachnid fossils to be found in Paleozoic strata (Fig. 34). The Silurian
scorpions appear to have lived in the water, since their fossils have gills, but by the
Carboniferous scorpions with such features are no longer found – fossils from the
Pennsylvanian age Mazon Creek beds have book lungs covered by protective
plates, and so were probably land-dwellers. The best scorpion fossils come from
Figure 34. Scorpions
119
thorax and abdomen, and the process and condition of fusion is called tagmosis. The
body is covered with an exoskeleton made up primarily of the protein chitin; lipids,
other proteins, and calcium carbonate also play a role. Primitively, each body seg-
ment bears a pair of segmented (jointed) appendages; in all living arthropods, many
of these appendages are dramatically modified or even lost. Arthropods generally
grow by molting their exoskeletons in a process called ecdysis. Movement of ap-
pendages is controlled primarily by a complex muscular system, divided into smooth
and striated components as in chordates. Cilia are not present. Most arthropods have
a pair of compound eyes and one to several simple («median») eyes or ocelli; either
or both kinds of eyes may be reduced or absent in some groups. Arthropods are
eucoelomate with the coelom formed by schizocoely, but the volume of the coelom
is much reduced and usually restricted to portions of the reproductive and excretory
systems. Most of the body cavity is an open «hemocoel,» or space filled loosely with
tissue, sinuses, and blood. The circulatory system is open and consists of a heart,
arteries, and the open spaces of the hemocoel. The gut is complete. Respiration
takes place through the body surface, and/or by means of gills, tracheae, or book
lungs. The nervous system is annelid-like, with a brain (=cerebral ganglion) and a
nerve ring surrounding the pharynx that connects the brain with a pair of ventral
nerve cords. These cords contain numerous ganglia. Most arthropds are dioecious
and have paired reproductive organs (ovaries, testes). Fertilization is internal in most
but not all groups. Most lay eggs, and development often proceeds with some form
of metamorphosis.
Subphylum Chelicerata
Class Merostomata (horseshoe crabs, eurypterids)
Class Pycnogonida (sea spiders)
Class Arachnida (spiders, ticks, mites)
Subphylum Crustacea
Class Remipedia
Class Cephalocarida
Class Branchiopoda (fairy shrimp, water fleas, etc.)
Class Maxillopoda (ostracods, copepods, barnacles)
Class Malacostraca (isopods, amphipods, krill, crabs, shrimp, etc.)
Subphylum Uniramia
Class Chilopoda (centipedes)
Class Diplopoda (millipedes)
Class Insecta
Crustacea
Crustaceans are members of the phylum Arthropoda. They are primarily marine, but
many also inhabit freshwater and terrestrial habitats from the deep-sea to the highest
mountain lakes. More than 52,000 species of crabs, shrimps, lobsters and their close
relatives have been described; that figure is twice the number of all amphibians, reptiles,
118
Plasmodium malariae
Introduction
Plasmodium malariae occurs mainly in the subtropical and temperate areas where
P. falciparum and P. vivax occur. However it is less frequently seen, responsible for
approximately 7% of all malaria in the world. It has an incubation period of 18-40
days and a periodicity of 72 hours.
Life cycle
The features of the life cycle, which are specific to P. malariae, are as follows:
a) Infected erythrocytes are not larger than uninfected ones and sometimes even
smaller.
b) Mature erythrocytes are attacked and rarely reticulocytes, so that the density
of parasites is very low; about 0.2% of erythrocytes are parasitised.
c) It is often difficult to distinguish between a large trophozoite and an immature
gametocyte.
Morphology
Parasites of P. malariae are typically compact heavily pigmented parasites, which
are usually smaller and more deeply stained than normal. They tend to parasitise
small, old red blood cells, they do not contain any inclusion dots and the parasitaemia
is usually low.
Trophozoites
Trophozoites are found as fairy large fleshy rings with a single chromatin dot.
These can be very distorted and can often take the form of bands across the cell. All
trophozoites have a single chromatin dot and contain pigment.
Gametocytes
Gametocytes contain large amounts of black pigment, with chromatin present as
a compact mass in females and diffuse in males. They occupy less than two thirds of
the red blood cell.
Schizonts
Schizonts are usually few in numbers with 6-12 large merozoites in a single ring.
Pigment is usually present as a central black mass. The parasites present are generally
only found at one stage of schizogony development.
Clinical Disease
Symptoms include headache, photophobia, muscle aches and pains, anorexia,
nausea and vomiting. Plasmodium malariae, like P. vivax and P. ovale are relatively
benign. However, chronic infections in children may lead to nephrotic syndrome
due to immune complexes depositing on the glomerular wall.
Diagnosis of malaria parasites
Introduction
The definitive diagnosis of malaria infection is still based on finding malaria
parasites in blood films. In thin films the red blood cells are fixed so the morphology
47
of the parasitised cells can be seen. Species identification can be made, based upon
the size and shape of the various stages of the parasite and the presence of stippling
(i.e. bright red dots) and fimbriation (i.e. ragged ends). However, malaria parasites
may be missed on a thin blood film when there is a low parasitaemia. Therefore,
examination of a thick blood film is recommended. With a thick blood film, the red
cells are approximately 6-20 layers thick which results in a larger volume of blood
being examined.
Thick Blood Films
In examining stained thick blood films, the red blood cells are lysed, so diagnosis
is based on the appearance of the parasite. In thick films, organisms tend to be more
compact and denser than in thin films.
Field’s stain method for Thick blood films
The method recommended for staining thick blood is Field’s Stain, which is
made from 2 components. Field’s A is a buffered solution of azure dye and Field’s B
is a buffered solution of eosin. Both Field’s A and B are supplied ready for use by the
manufacturer.
Method
1. Place a drop of blood on a microscope slide and spread to make an area of
approximately 1 cm2. It should just be possible to read small print through a thick film.
2. The film is air dried and NOT fixed in methanol.
3. The slide is dipped into Field’s stain A for 3 seconds.
4. The slide is then dipped into tap water for 3 seconds and gently agitated.
5. The slide is dipped into Field’s stain B for 3 seconds and washed gently in tap
water for a few seconds until the excess stain is removed.
6. The slide is drained vertically and left to dry.
Microscopic Features of the Field’s stained thick blood film
a) The end of the film at the top of the slide when it was draining should be
looked at. The edges of the film will also be better than the centre, where the film
may be too thick or cracked.
b) In a well-stained film the malaria parasites show deep red chromatin and pale
blue cytoplasm.
c) White cells, platelets and malaria pigment can also be seen on a thick film.
The leucocyte nuclei stain purple and the background is pale blue. The red cells are
lysed and only background stroma remains. The occasional red cell may fail to lyse.
d) Schizonts and gametocytes, if present, are also easily recognisable.
e) A thick film should be examined for at least 10 minutes, which corresponds to
approximately 200 oil immersion fields, before declaring the slide negative.
N.B.
a) As a result of haemolysis of the red blood cells due to staining of an unfixed
film, the only elements seen are leucocytes and parasites, the appearance of the
latter being altered. Consequently:
b) The young trophozoites appear as incomplete rings or spots of blue cytoplasm
with detached chromatin dots.
48
Occasionally, if the stool specimens are several hours or 1-2 days old, eggs may
develop to more advanced stages. Ascaris eggs usually have only 1 cell when passed
in the faeces; however, the single cell may divide and, in old specimens, eggs with 2
or 4 cells may be seen. Hookworm eggs in specimens that are several hours old may
contain 16, 32 or more cells. In 12 – 24 hours, the egg may be embryonated and later
still the larvae may hatch. Therefore, when observing the stage of development of
helminth eggs, be sure that the stool specimen is freshly passed. If it is several hours
or a day old, expect to see changes in the stage of development of some species.
Ideally only fresh samples should be accepted for diagnosis.
4. Thickness of the egg shell: Some species, like Ascaris, have thick shells; others,
like hookworm, have thin shells.
5. Colour: Some eggs are colourless (e.g., hookworm, Enterobius), others are
yellow or brown (Ascaris, Trichuris).
6. Presence of characteristic like opercula (lids), spines, plugs, hooklets, or
mammillated outer coats.
TOPIC: ARTHROPODA, ARACHNOIDEA, ACARINA
Phylum Arthropoda
Arthropods are said to be the most successful phylum. It includes about 80% of
all species of animals with well over one million species known, although it’s thought
that over six million could exist. This phylum includes the most diverse species in
terms of distribution, being found in water, on land, and in the air, and it includes the
highest number of individuals within each species. They are found in a variety of
environments, from mountain tops to deep oceans, and are able to survive tremendous
environmental conditions. There are several features characteristic of all arthropods,
including a hardened exoskelton, specialized segments, jointed appendages, modified
mouthparts, and specialized respiratory structures such as gills, trachea, or book
lungs. The sexes are usually separate. The group includes a variety of feeding forms:
herbivores, carnivores, and filter feeders to name a few.
Arthropods include an incredibly diverse group of taxa such as insects,
crustaceans, spiders, scorpions, and centipedes. There are far more species of
arthropods than species in all other phyla combined, and the number of undescribed
species in the largest assemblage of arthropods, the insects, probably numbers in the
tens of millions. Members of the phylum have been responsible for the most
devastating plagues and famines mankind has known. Yet other species of arthropods
are essential for our existence, directly or indirectly providing us with food, clothing,
medicines, and protection from harmful organisms.
A number of important characteristics are shared by most members of this phylum.
Arthropods are bilaterally symmetrical protostomes with strongly segmented bod-
ies. Segmentation affects both external and internal structure. Some segments are
fused to form specialized body regions called tagmata; these include the head,
117
 If the microfilariae is ingested by an appropriate species of Cyclops, they break c) The stippling of P. vivax and P. ovale may be less obvious although occasion-
though the soft mid-intestine wall and come to lie in the body cavity. The larvae un- ally ghost stippling may be seen.
dergo two moults and become infective in approximately 3 weeks. Humans become d) The cytoplasm of late trophozoites of P. vivax and P. ovale may be fragmented.
infected by accidentally ingesting through drinking water the infective Cyclops. Upon e) Caution should be exercised when examining thick blood films as artefacts
ingestion the larvae are activated to penetrate through the gut wall, and migrate through and blood platelets may be confused with malaria parasites.
the tissues, moulting twice and finally becoming lodges in the viscera or subcutaneous Thin Blood Films
tissues. Maturation of the worms is slow taking about 1 year to reach sexual maturity When examining thin blood films for malaria you must look at the infected red
before the females are ready to migrate to the skin to release their larvae. blood cells and the parasites inside the cells.
Morphology 1. Rapid Field’s stain for thin films
The adult female worm measures up to 1 metre in length whereas the male This is a modification of the original Field’s stain to enable rapid staining of
measures about 2cm. fixed thin films. This method is suitable for malaria parasites, Babesia sp., Borrelia
Clinical disease sp. and Leishmania sp.
After ingestion of the Cyclops, there is no specific pathology associate with the Method
mucosal penetration and larval maturation in the deep connective tissues. Erythema 1. Dry the film
and tenderness can be associated with blister formation. The patient can also exhibit 2. Fix in methanol for 1 minute.
vomiting, diarrhoea, asthmatic attacks. Symptoms usually subside when the lesion 3. Flood the slide with 1 ml of Field’s stain B, diluted 1 in 4 with distilled water.
erupts. If the worm is removed, healing usually occurs without any problems. If the 4. Immediately, add an equal volume of undiluted Field’s stain A, mix well and
worm is damaged or broken during removal, there may be intense inflammatory allow to stain for 1 minute.
reaction with possible cellulitis along the worms migratory tract. This can result in 5. Rinse well in tap water and drain dry.
arthritis and synovitis. Uses
Laboratory diagnosis This is a useful method for rapid presumptive species identification of malarial
The best remedy for removing the adult worm is a slow process of daily gently parasites. It shows adequate staining of all stages including stippling (mainly Maurer’s
rolling the worm around a small stick and slowly pulling it out of the skin. With this dots). However, staining with Giemsa is always the method of choice for definitive
method you must be careful not to pull apart the worm as it will recoil back into the species differentiation.
skin and cause secondary infections.
At this moment in time this parasite is being effectively controlled due to a strict 2. Giemsa stain for thin films.
control program. The program includes stopping people from drinking infected water, Method
putting muslin over the water jars, which they use to collect the water in, thus 1. Air dry thin films
preventing the cyclops from being collected in the water. Educating the communities 2. Fix in methanol for 1 minute
about the parasite and adding temphos to the water thus killing off any microfilariae 3. Wash in tap water and flood the slide with Giemsa diluted 1 in 10 with buffered
in the water. distilled water pH 7.2. The diluted stain must be freshly prepared each time.
Identifying Intestinal Helminths 4. Stain for 25-30 minutes.
The usual diagnostic stages for identifying medically important helminths are 5. Run tap water on to the slide to float off the stain and to prevent deposition of
the eggs and larvae. Occasionally, adult worms like Ascaris and Enterobius may be precipitate on to the film. Dry vertically.
seen and segments or proglottids are used for diagnosing certain tapeworms. 6. Examine the film using the x100 objective.
If an egg is found the following features as described should be carefully observed Microscopic features of the thin blood film
in order to make a specific identification. 1. Examine the tail end of the slide where the red cells are separated into a one-
1. Size: The length and width are measured and are generally within a specific cell-layer thick.
range. 2. An alkaline buffer pH 7.2 is vital for clear differentiation of nuclear and
2. Shape: Each species has its own particular shape. cytoplasmic material and to visualise inclusions such as Schüffner’s / James’s dots
3. Stage of development when passed: In some species, the eggs consist of a in the red cells. Acidic buffer is unsuitable.
single cell; in some, there may be several cells; and some species are usually em- 3. The red cells are fixed in the thin film so the morphology of the parasitised
bryonated (i.e., they contain a larva) when passed in the faeces. cells and the parasites can be seen.
116 49
 4. On a well stained film the chromatin stains red/purple and the cytoplasm blue. The life cycle usually involves copepod intermediate host. They are parasitic in the
Leucocytes have purple nuclei, the red stippling, if present should be clearly visible. connective tissue or coelom of vertebrates. The disease associated with this parasite
Rings of the four main species of malaria may look alike. If you see rings, look is known as Dracunculiasis (Fig. 33).
for older stages. Patients with a P. falciparum infection only, rings are usually seen; Mature female worms, which are gravid with microfilariae, migrate to the
older stages are present only in severe infections. superficial layers of skin of humans, especially those regions, which are most likely
In poorly stained slides, Schüffner’s dots may not be visible, so it is essential that to come in contact with water, such as the ankle, foot, arms and shoulders. Here the
correct staining methods are used. Also Schüffner’s dots may not be seen in the worms secrete a substance (substance is unknown), which causes a blister to rise
earlier rings of P. vivax or P. ovale. over its anterior end where it has pierced the lower layers. The blister eventually
forms into an ulcer which on contact with water, the uterus is projected out of the
Estimation of Percentage Parasitaemia of Plasmodium falciparum ulcer cavity, and a cloud of milky white secretion, containing hundred of active
larvae, is released. Once out of the water again the uterus dries and shrivels prevent-
Counting of red blood cells infected with parasites of P. falciparum is essential ing the release of further larvae.
and the percentage parasitaemia should always be reported as this has implications
for prognosis and the pattern of treatment employed.
The recommended procedure for estimating the percentage parasitaemia in a
thin blood film is by expressing the number of infected cells as a percentage of the
red blood cells e.g. 3 parasitised red cells / 100 red blood cells or 3% parasitaemia.
A red blood cell infected with multiple parasites counts as one parasitised red cell.
The percentage parasitaemia should be calculated by counting the number of
parasitised red bloodcells in 1000 cells in a thin blood film.
Method 2
Alternatively the World Health Organisation recommend a method, which compares
the number of parasites in a thick blood film with the white blood cell count.
The parasitaemia is estimated by first counting the number of parasites per 200
white blood cells in a thick blood film and then calculating the parasite count / ml
from the total white blood cell count / ml.
Knowledge of either % parasitaemia or total parasite count is essential for the
correct clinical management of P. falciparum malaria.
The Intersep Malaria One Step Rapid tests are qualitative membrane based assays
for the detection of malaria antibodies in serum, plasma or whole blood.
Separate devices are available for the detection of Plasmodium falciparum and
Plasmodium vivax antibodies.
The device is precoated with the appropriate Malaria antigen on the test line and
anti-conjugate antigen on the control line.
During the test the sample is exposed to the conjugate complex. The sample
migrates up the membrane binding with the test line in the presence of the malaria
antibody. Independent of a positive reaction the sample continues to migrate to the
control line where the conjugate binds to the membrane bound anti-conjugate antigen
to demonstrate correct device function.
A positive result is indicated by two lines and a negative result by the sole presence
of the control band.
Thin blood films for malaria diagnosis are best prepared from venous or
capillary blood taken directly from the patient, without the addition of anticoagulant. Figure 33. Dracunculus medinensis
50 115
 Mansonella streptocerca However, this is not usually possible in a clinical laboratory, as many samples are
received from general practices and other hospitals. All anticoagulants have some
Microfilaria of M. streptocerca were first reported in the skin of a West African effect on the morphology of malaria parasites and the red blood cell they inhabit.
patient in 1922. These microfilaria are primarily found in the skin but have been also This effect depends on the stage of the parasite, the time taken for the blood to
reported in the blood. This species occurs in Ghana, Cameroon and Zaire. The adults reach the laboratory and the type of anticoagulant used. If it is necessary to use an
are poorly known, and occur in the cutaneous tissue of man and chimpanzee. anticoagulant, the films should be prepared as soon as possible after the blood has
The microfilariae do not exhibit periodicity with the intermediate hosts being been taken. If the films cannot be made immediately, potassium EDTA is the
Culicoides grahamii and possibly other Culicoides species. anticoagulant of choice. However if the blood is left for several hours in EDTA,
Life cycle the following effects may be seen.
The life cycle is the same as that of the blood Mansonella species.
Clinical Disease Table №2
Infection is characterised by pruritic dermatitis and hypopigmented macules. Differential diagnostic features of human Plasmodia species – Giemsa stained thin
Laboratory diagnosis film of peripheral blood
Mansonella streptocerca can be diagnosed by demonstrating the microfilaria in
a skin snip. Snips are collected from several sites, usually the shoulders and buttocks
and sometimes the chest and calves. The snips are placed immediately in 0.5ml of
0.9% sodium chloride in a microtitre plate and left for 4 hours to allow the microfilaria
to migrate out of the tissues. After 4 hours, the wells are examined using an inversion
microscope. The microfilaria should still be moving and can be identified by staining
with Giemsa at pH 6.8
Small, thin, microfilaria. Does not have a sheath. Nuclei extend to end of tail.
Tail is hooked; its tip is rounded or forked. Found only skin.
Table 9
Differential features of Onchocerca volvulus and Mansonella streptocerca

Dracunculus medinensis

Introduction
Dracunculus medinensis is a non-filarial parasite as it only has one uterus whereas
filaria have two. It is usually associated with places where there is a lack of clean
drinking water e.g. step wells in India, covered cisterns in Iran, and ponds in Ghana.
114
1. Sexual stages may continue to develop and male gametocytes can exflagellate,
liberating gametes into the plasma. These can be mistaken for organisms such as
51
Borrelia. Gametocytes of P. falciparum, which have a characteristic crescent shape,
may round up and then resemble those of P. malariae.
2. Acole forms, which are characteristic of P. falciparum, may be seen in P. vivax
because of attempted re – invasion of the red blood cell by merozoites.
3. Mature trophozoites of P. vivax may condense when exposure becomes
prolonged and in cases of extreme exposure, red blood cells containing gametocytes
and mature schizonts may be totally destroyed along with the contained parasites.
The malaria pigment, haemozoin, always remains and can provide a clue to the
presence and, to an expert eye identity of the parasite.
4. The morphology of the red blood cell may be altered by shrinkage or crenation.
The Ciliates
The ciliates belong to the family Ciliophora. They possess simple cilia or
compound ciliary organelles, 2 types of nuclei and a large contractile vacuole. The
only member of the ciliate family to cause human disease is Balantidium coli.
Balantidium coli
Introduction
Balantidium coli is widely distributed in warmer climates, which is where human
infections most commonly occur. The organisms inhabit the large intestine, caecum
and terminal ileum where they feed on bacteria. The most common hosts being
humans, pigs and rodents. Human infection is usually from pigs and is rare. Figure
12 illustrates the life cycle of Balantidium coli.
Morphology of cyst
The cyst is spherical or ellipsoid and measures from 30-200mm by 20-120mm.
It contains 1 macro and 1 micronucleus. The cilia are present in young cysts and may
be seen slowly rotating, but after prolonged encystment, the cilia disappear. Cysts
form when diarrhoea subsides and the rectal contents become formed. The cyst,
ingested by a fresh host, excysts to liberate the trophozoite. Diagram 2 illustrates a
B. coli cyst.
Morphology of trophozoite
Trophozoites of B. coli measure approximately 30-150mm in length x 25-120mm
in width but have been known to attain lengths of up to 200mm. They are oval in
shape and covered in short cilia. A funnel shaped cytosome can be seen near the
anterior end. Multiplication is by binary fission in the trophozoite stage. In an
unstained preparation, the organisms are easily recognised because of their size and
rapid revolving rotation. In a stained preparation, the characteristic macro and
micronuclei may be observed.
Clinical Disease
Severe B.coli infections may resemble amoebiasis. Symptoms include diarrhoea, nausea,
vomiting and anorexia. The diarrhoea may persist for long periods of time resulting in
52
Clinical Disease
Clinical manifestations are due to microfilarie in the epidermis.
Light infections may be asymptomatic or cause pruritis. This leads to scratching
which can result in infection. Lyphadenopathy may also be a feature of early infection.
After months or years, onchodermatitis results in secondary stage of thickening due
to intradermal oedema and pachydermis. There is a loss of elastic fibres resulting in
hanging groin, hernias and elephantiasis of the scrotum. There is finally atrophy of
the skin resulting in loss of elasticity. There is mottled depigmentation of the skin.
Ocular lesions are related to the intensity of the microfilariae in the skin. Ocular
lesions include sclerosing keratitis, secondary glaucoma and cataract, coroidoretinitis
and fluffy corneal opacities. The major complication of onchocerciasis is the
development of lesions in the eye, which may result in blindness or other distressing
ocular diseases.
Laboratory diagnosis
1. Analysis of Skin Snips
Small amounts of skin are collected by using a needle to raise the skin and then
to slice about 1 mg of skin to a depth of 0.5mm. Snips are collected from several
sites, usually the shoulders or the buttocks and sometimes the chest and calves. The
snips are placed immediately in 0.5ml normal saline in a microtitre plate and left for
4 hours to allow the microfilariae to migrate out of the tissues. After 4 hours, the
wells are examined using an inversion microscope. The microfilariae should still be
moving and can be identified from the table below. The microfilariae can also be
collected by filtration or centrifugation and the deposit containing microfilariae can
be stained with Giemsa at pH 6.8.
2. Analysis of Biopsies
Biopsies of tissue nodules can be dabbed on to a slide to produce impression
smears and then stained with Giemsa stain at pH 6.8 for the presence of microfilariae.
Recent advances in diagnostic methods includes and ELISA-based antibody
detection assay which utilises a cocktail of recombinant antigens. The advantages of
using this test are that it is highly sensitive (almost 100% in onchocerciasis foci). It
is also highly specific (100%), it also uses finger prick blood. Therefore, reducing
the painful procedure of gaining a skin snip.
The disadvantages is that it requires advanced ELISA apparatus and reagents
and cannot distinguish between past and present infections due to it detecting
antibodies which stay present in the body for a long time after the infection. Another
modern detection method is for Parasite DNA detection, which is based on the
amplification of specific DNA sequences form microfilariae using molecular biology
technology. The advantages of this technique are its exquisite sensitivity and detect
active infections only. The disadvantages are that it requires specialised equipment
and expensive reagents. Also it still requires a skin snip but a urine assay is a possibility
for the future.
Thick microfilaria. Does not have a sheath. Head often spatulate. Nuclei do not
extend to tip of tail. Found only in skin.
113
 Figure 32. Onchocerca volvulus

The microfilariae of O. volvulus are unsheathed and are usually found in the dermis.
They measure between 221-287mm long. Figure 12. Balantidium coli
112 53
acute fluid loss. Balantidium coli also has the potential to penetrate the mucosa resulting in
ulceration just as those of Entamoeba histolytica, but perforation is more common. Metastatic
lesions do not occur. Extra-intestinal disease has also been reported, but is rare.
Laboratory Diagnosis
Wet preparations of fresh and concentrated stool samples reveal the characteristic
cysts and motile trophozoites. They are easier to identify in direct-smear saline
preparations than permanently stained faecal smears.
TOPIC: PLATHELMINTHES, TREMATODA
Helminth Parasites
The word “worm” is used loosely to describe organisms with elongated bodies
and a more or less creeping habit.
The word “Helminth” does mean “worm”, but in zoological terms it is more
restricted to members of the phyla Platyhelminths, Nematoda and Acanthocephala.
There are three groups of medically important helminths; Cestodes (tapeworms),
Nematodes (roundworms) and Trematodes (flukes).
These parasites live in both the body spaces (gut lumen, bile ducts, lungs, oral
cavity, etc.) and in tissues (blood, muscles and skin).
The Trematodes
The trematodes (or flukes) are leaf shaped and can outer cover called the tegument,
which may be smooth or spiny. There are 2 suckers or attachment organs, an anterior
oral sucker and a posterior ventral sucker. The suckers form a characteristic feature
of the group, from which the name Trematode is derived from (trematodes is a Greek
word for meaning holes). They can occur in a variety of host environments, with the
majority being endoparasitic but some are found to be ectoparasitic.
Most trematodes are hermaphroditic and most of their body consists of the
reproductive organs and their associated structures. The digestive system is well
developed; they generally feed on intestinal debris, blood, mucus and other tissues,
depending on the host environment.
Trematodes require an intermediate host in their life cycle with vertebrates being
the definitive host. Larval stages may occur in either invertebrate or vertebrate hosts.
There are three groups of trematodes: the Monogenea, which typically are external
parasites of fish with direct life cycles. The Aspidogastrea, these are endoparasites
with the entire ventral surface as an adhesive organ. Finally the third group is the
Digenea, these are endoparasites with simpler adhesive organs and life cycles
involving one or more intermediate hosts (indirect life-cycle). This section
concentrates on the Digenean trematodes.
Most digenean trematodes inhabit the alimentary canal of vertebrates and many
of the associated organs, such as the liver, bile duct, gall bladder, lungs, bladder and
54
Microfilaria worms found in tissue and skin
The main species of microfilariae found in the skin and tissue are Onchocerca
volvulus and Mansonella streptocerca. Microfilariae of Onchocerca volvulus and less
often, Mansonella streptocerca migrate through the dermis causing itching and skin
texture changes and occasionally arrive in the eye where they cause blindness. Detection
of these microfilariae is from skin snips or nodule biopsies. When high numbers of
microfilariae are present, they can occasionally be found in the blood and urine.
Onchocerca volvulus
Introduction
Onchocerca volvulus is mainly found in West Africa and Central and South America.
Onchocerciasis, also known as river blindness, is a major public health problem,
especially in West Africa; there an eradication program has been established. It is one
of the world’s most distressing diseases of helminth origin, often resulting in blindness.
Onchocerca volvulus is transmitted by the species Simulium or black fly whose breeding
habitat is by fast flowing rivers or streams, therefore there is a patchy distribution of
the disease as it is specified to where water courses are. The adult worms are found in
nodules or onchodermata in superficial sites, but may invade other tissues.
It is estimated that there are 18 million cases worldwide with 17.5 million being
found in Africa. Nigeria is the most infected region. The rate of morbidity is high in
relation to those with an infection.
The life cycle is similar to W. bancrofti, except that the intermediate hosts are
various species from the genus Simulium (Black flies), the most important species is
Simulium damnosum (Fig. 32).
The microfilariae are ingested by a Black fly during a blood meal, from where
they are carried to the midgut where they penetrate the epithelium and migrate, via
the haemocoele, to the indirect flight muscles. Here they undergo two moults,
L1 – L3 and develop into infective L3 larvae which move to the mouth parts.
Development is completed in 6-9 days.
When the infected fly takes another blood meal the infective larvae are once again
transmitted into another host (definitive host). The microfilariae are released from the
mouth parts and transmitted directly into the hosts bloodstream. Moulting takes place
form L3 – L4 within 2-5 days and the larvae then migrate widely through the body under
the skin and between muscles, ligaments and tendons. The final moult to L5 occurs at
1.5-2.5 months after transmission. Male worms are known to mature in about 4 months
later. Female worms initiate the formation of the nodules and the males may join later.
The sexually mature female worms release microfilariae, which migrate out from the
nodules into the skin and other tissues, most significantly into the eye.
Morphology
The whitish adult worm lies coiled within capsules in the fibrous tissue. The
female can measure up to 50cm while the males are shorter measuring up to 5 cm.
111
 Mansonella species
Introduction
Members of the genus Mansonella are filarial nematodes, which rarely cause
serious disease. However, they can be found in geographical areas where Wuchereria
bancrofti, Loa loa and Onchocerca volvulus also occur and therefore must be
differentiated from these pathogenic microfilariae. Unlike the pathogenic blood
filariae, they do not exhibit periodicity.
Life cycle
There is a general life cycle for the Mansonella species of filarial nematodes.
The microfilaria are picked up by the vector Culicoides sp. (biting midges) during a
blood meal. The larvae develop within the body of the Culicoides sp. and are re-
introduced into the human host when the vector takes another blood meal. They are
found in various sites around the human host body.
Table 8
Comparison of the main human filarial nematodes
110
ureter. These organs are rich in cavities containing food such as blood, mucus, bile
and intestinal debris.
The digenean trematodes have a complex life cycle, with rare exceptions, always
involve a mollusk host. There may be six larval stages – the miracidium, sporocyst,
redia, cercaria, mesocercaria (rare) and the metacercaria (the majority have 4 or 5 stages).
Trematode eggs have a smooth hard shell and the majority of them are operculate.
Intestinal and Liver Flukes: Gastrodiscoides hominis,
Nanophyetus salmincola,
Fasciolopsis buski, and Fasciola hepatica
Classification. Helminths. Phylum Platyhelminthes. Trematodes.
Diseases
Paramphistomiasis (Gastrodiscoides). Nanopbyetus infection, fasciolopsiasis
(Fasciolopsis). Fascioliasis or sheep liver fluke infection (Fasciola).
Geographic Distribution
Gastrodiscoides is a parasite of pigs and humans in India, Southeast Asia, and
parts of the former USSR; N. salmincola is found in North America. Fasciolopsis
occurs in China, Taiwan. Thailand, Indonesia, India. Bangladesh, and possibly
Cambodia. Myanmar and Vietnam. Fasciola hepatica has a worldwide distribution,
especially in sheep-raising countries.
Location in Host
Gastrodiscoides occurs in the cecum and ascending colon; both Nanophyetus
and Fasciolopsis live attached to the wall of the small intestine; and Fasciola lives
in the bile ducts of the liver.
Morphology. Adult Worms
Gastrodiscoides adults are thick-bodied, pinkish in color, and measure 5-14 mm
long by 5-8 mm in width. The body is pyriform with a small, conical anterior portion
and a larger, discoidal posterior part that is concave ventrally. Adults of Nanophyetus
are small, pyriform flukes measuring 0.8-1.1 mm in length.
Fasciolopsis buski adults have a large, broad, fleshy hotly measuring 20-75 mm long
and 20 mm wide. The anterior end is rounded, not cone-shaped. Although the reproductive
organs are extensively branched, the esophagus and intestinal ceca are not. The smaller
branched ovary is situated anterior to the paired, large, dendritic testes.
Fasciola hepatica adults are large, broadly flattened, fleshy worms measuring up
to 30 mm long and 13 mm wide. The anterior end is distinctly cone-shaped. All of the
internal organs, including the esophagus, intestinal ceca, and reproductive organs, are
extensively branched. The paired testes lie one behind the other, posterior to the ovary.
Eggs
Unembryonaled eggs of Gastrodiscoides passed in feces measure 130-160 µm.
long by 62-75 µm in width and have a pale greenish-brown color. Eggs of Nanophyetus
are light brown in color, broadly ovoid, and unembryonated when discharged in
55
feces. They measure from 64-97 µm long by 34-55 µm wide, and have a bluntly
pointed, thickened, and darkened area of the shell at the abopercular end. Eggs of
Fasciolopsis and F. hepatica are similar in size and appearance, and are difficult to
distinguish from each other. The operculated eggs are large, a light yellowish-brown,
broadly ellipsoidal, and unembryonated when excreted in feces. They measure 130-
150 µm by 63-90 µm. The operculum is small and indistinct. At the abopercular end
of the shell of Fasciola eggs there is often a roughened or irregular area that is not
seen in Fasciolopsis eggs.
Life Cycles
Although details of the Gastrodiscoides life cycle have not been clarified, it is
believed that cercariae liberated from the snail intermediate host encyst to the
metacercarial stage on aquatic vegetation. Nanophyetus salmincola infection is
acquired by ingestion of infected fish that serve as the second intermediate host.
The life cycles of Fasciolopsis and F. hepatica are similar (Fig. 13). Unembryonated
eggs pass in feces into water where they become embryonated after 2 weeks or longer
and infect appropriate snails. Cercariae emerge from these snails, attach to aquatic
vegetation, such as watercress and water caltrop nuts, and undergo encystation to the
metacercarial stage. Infection is acquired by ingestion of the metacercariae on this
vegetation. Fasciolopsis buski matures directly in the duodenum in approximately 3
months. Larval F. hepatica migrate through the intestinal wall into the abdominal
cavity, enter the liver, and burrow through the parenchyma to enter the bile ducts
where the adult worms reach maturity in 3 to 4 months. It is not uncommon for these
worms to end up in ectopic locations, including the abdominal wall, lungs, brain, and
orbit because of the extraintestinal migration of the larval stages of Fasciola.
Animals play a significant role in the life cycles of these flukes. Swine are the
usual host for Gastrodiscoides. Many mammals and even birds are infected with
Nanophyetus. In canids, an often fatal rickettsial disease known as salmon poisoning
is carried by these flukes; this is particularly prevalent in the northwest United States
and develops in dogs who have eaten raw infected salmon or other fish. Pigs are an
important reservoir host for Fasciolopsis infection. Sheep and cattle are important
hosts for F. hepatica infection, and in endemic areas economic losses in these animals
may be severe.
Diagnosis. Detection of characteristic eggs in feces.
Diagnostic Problems
Human Gastrodiscoides infection is uncommon but the large size of the eggs
should aid in diagnosis. Nanophyetus salmincola infection principally has been
reported from the northwestern United States, although people from eastern Siberia Figure 31. Loa loa
have a king history of infection with a different species of this parasite.
Infection is associated with eating raw, incompletely cooked or smoked salmon Laboratory diagnosis
and steelhead trout. Eggs of Nanophyetus must be distinguished from those of Kinked and sheathed microfilaria. Sheath does not stain with Giemsa stain; does
Diphyllobothrium latum, which they somewhat resemble in size and morphology. stain with haematoxylin stains. Nuclei crowded extending to tip of tail; tip of tail
The similarity in size and morphology of the eggs of F. hepatica and Fasciolopsis tapers. Cephalic space as long as it is broad. Innerbody does not usually stain. Found
may lead to difficulties in arriving at a correct diagnosis in areas of southeast Asia in blood.
56 109
 Loa loa

Introduction
Loa loa, also known as the African eye worm, is a filarial nematode endemic in
the rain forests of West and Central Africa. It is transmitted by Chrysops species,
also known as mango flies or horse flies and humans are the only known reservoir. It
is estimated that 2-13 million humans are infected with the larvae.
Adults migrate in the subcutaneous tissues of man and monkeys, with them
eventually migrating across the eyeball under the conjunctiva.
Life cycle
The adult worms live in the subcutaneous and deep connective tissues and the
microfilariae are found in the peripheral blood, where they can be in ingested by
the Crysops fly (day biting fly). The adults can live in the tissues for up to 17 years
(Fig. 31).
Once the microfilariae have been taken up by the Chrysops during a blood meal
they develop within the fat boduy. They develop through to L3 within 10-12 days.
The microfilariae, L3 re-enter the host’s blood stream when the fly takes another
blood meal. They reach adult worms within 4- 6 months living in the subcutaneous
and deep connective tissues.
The microfilariae exhibit diurnal periodicity, the highest numbers being detected
in blood between 10am and 2pm.
Morphology
Adult males of Loa loa are 2-3.5cm long and the females from 5-7cm. The
microfilariae of Loa loa are 250-300mm. They possess a sheath, which stains blue-
grey with Delafield’s haematoxylin. The sheath does not stain with Giemsa. The tail
gradually tapers to a rounded end, the densely packed nuclei extending to the tip.
Clinical disease
Many patients infected with Loa loa appear to be asymptomatic and the migration
of the adult worm through the subcutaneous tissues often goes unnoticed, unless
passing beneath the conjunctiva of the eye. They can be seen crossing the eye, but it
is a rapid process taking approximately 15-20 minutes. Hypereosinophilia and
increased antibody levels, especially IgE are also noted. Eyeworm episodes are as
equally common in man as well as women with common re-occurrences. There is an
increased incidence with age.
The most common pathology associated with Loa loa infections are Calabar
swellings, which are inflammatory swellings resulting in a localised subcutaneous
oedema. These swellings are due the host’s response to the worm or its metabolic
products and can be found anywhere in the body but most commonly in the Figure 13. Fasciola hepatica
extremities. These swellings last from 1-3 days. They develop rapidly and last one
to three days, usually accompanied by localised pain, urticaria and pruritis. There is where these species overlap in geographic distribution. In such instances, clinical
a higher frequency of calabar swellings in women with common re-occurences. evaluation of the patient may be an important aid in diagnosis. Whereas F. hepatica
Serious complications such as cardiomyopathy, encephalopathy, nephropathy is worldwide in distribution, a related species that infects herbivorous animals, Fasciola
and pleural effusion have been recorded. gigantica, can be found in the liver of humans in southeast Asia and Africa. The eggs
108 57
of F. gigantica typically are larger (160-190 µm long by 70-90 µm in width) than those
of F. hepatica and Fasciolopsis. The migration of F. hepatica and F. gigantica to
ectopic locations in humans may be a feature of these infections that can pose a
diagnostic problem. Spurious infections of F. hepatica and F. gigantica might be noted
in individuals who have eaten the liver of infected cattle, water buffalo, or sheep.
Comments
Patients with Nanophyetus infection typically present with abdominal discomfort,
diarrhea, and unexplained peripheral blood eosinophilia. Most infections with
Fasciolopsis are light and asymptomatic, but in heavy infections, diarrhea and
epigastric pain simulating a peptic ulcer are seen. In F. hepatica infection, extensive
damage to the liver parenchyma because of the migration of immature flukes may
occur. Acute Fasciola infections in children can be fatal.
Table 3
Table describing the characteristics, which differentiate the various Fasciola
species, which are important to man
Introduction
Fasciola, Fasciolopsis and Echinostoma species are trematodes which parasitise
the liver and intestines of a variety of vertebrates, they are hermaphroditic.
Fasciola hepatica trematodes are not thought to infect man but in fact man is not
an unusual host, with infections being reported in many countries including Europe
and the USA. The eating of unwashed watercress appears to be the source of infection,
with them ending up in the liver. The most common host is sheep where they can
cause severe disease.
Fasciolopsis buski (giant intestinal fluke) is a duodenal parasite infecting both
man and pigs. They are found widespread in Asia and China, but they have been
found to be endemic in Taiwan, Thailand, Bangladesh and India. Night soil is used
as a fertilizer in these countries on plants such as water chestnut and caltrops. The
snails graze on these crops and also the definitive hosts eat them raw and unwashed,
peeling the edible water plants with their teeth.
58
This species like W. bancrofti also parasitises the lymph nodes and lymphatics;
the adults of the two species are indistinguishable. Causing Malayan filariasis.
Life cycle
The adult worm inhabits the lymphatics and the female produces sheathed
microfilariae, which circulate in the peripheral blood. The mosquito acquires the
infection by ingestion of the microfilaria in the blood meal. The microfilaria lose
their sheath on arrival in the stomach of the mosquito. The larvae migrate to the
thoracic muscles and develop into infective larvae over a period of 6-14 days. The
larvae then migrate to the mouth parts of the mosquito and enter the skin of the
definitive host through the puncture wound when a blood meal is taken. The
infective larvae enter the peripheral lymphatics where they grow to mature male
and female worms.
Morphology
The adult worms of B. malayi are smaller than those of W. bancrofti. The
microfilariae of Brugia malayi are 170-230mm in length and have 2 terminal nuclei
that are distinctly separated from the other nuclei in the tail. The last terminal nucleus
is quite small and is at the tip of the tail. The sheath stains deep purple with
haematoxylin stain.
Clinical disease
Clinical features of B. malayi are similar to those of W. bancrofti, however in
B. malayi, unlike Wuchereria bancrofti, genital involvement, hydrocele and chyluria
are rare.
Many patients are asymptomatic. Patients may present fever. Lymphaginitis and
lymphadenitis develop in the lower extremities. An imflammatory reaction occurs in
the lymphatic vessels that harbour the adult worms. Oedema develops which may
resolve after the first few attacks. However, in long standing disease after several
episodes of lymphaginitis, thickening and verrucous changes in the skin known as
elephantiasis.
Some patients with lymphatic filariasis do not exhibit microfilaraemia. However,
they do have high eosinophilia, high IgE level and high anti-filarial antibody titres.
Laboratory diagnosis
Kinked microfilaria. Sheath stains deep pink with Giemsa stain. Does stain
with haematoxylin stains. Nuclei crowded and fill the whole body. Empty space
between nuclei and body wall. Cephalic space twice as long as it is broad. Innerbody
may or may not stain; when it does, it is prominent. Found in blood.
Brugia Timori
Brugia timori is found in the islands of Indonesia and exhibits a strictly nocturnal
periodicity. The lifecycle and disease closely resembles that of Brugia malayi.
However, the microfilariae can be distinguished from those of B. malayi in that they
are about 310mm in length. The sheath satins pink with Giemsa and the nuclei at the
tip of the tail are similar to those of B. malayi
107
 The microfilariae are 230-275mm in length. The tail of the microfilariae of W.
bancrofti tapers to a delicate point and exhibits no terminal nuclei. The sheath the
microfilariae of W. bancrofti stains with haematoxylin stain.
Clinical disease
Many patients are asymptomatic. Patients may present with fever, lymphangitis
and lymphadenitis. Lymphangitis commonly affects the lower extremities and there
may also be genital and breast involvement. An inflammatory reaction occurs in the
lymphatic vessels that harbour the adult worms. Oedema develops which may resolve
after the first few attacks. A late complication resulting in thickening and verrucous
changes in the skin known as elephantiasis may occur after recurring lymphangitis.
Secondary bacterial and fungal infections may occur in patients with long-standing
elephantiasis.
Obstruction of the genital organs may result in hydrocoele formation and scrotal
lymphoedema. Obstruction of the retroperitoneal lymphatics may cause the renal
lymphatics to rupture into the urinary tract producing chyluria.
Some patients with filariasis do not exhibit microfilaraemia but develop tropical
pulmonary eosinophilia, which is characterised by peripheral eosinophilia, wheeze
and cough. High eosinophilia, high IgE level and high anti-filarial antibody titres are
features of this syndrome.
Laboratory diagnosis
Sheath may or may not stain with Giemsa; does stain with haematoxylin stains.
Discrete nuclei. Empty space between the nuclei and the body wall. No nuclei in tip
of tail. Innerbody is rarely visible in Giemsa. Does not stain with haematoxylin.
Cephalic space as long as it is broad. Tip of tail may be bent underneath the body.
Found in blood.
Brugia malayi
Introduction
Brugia malayi is a nematode causing lymphatic filariasis in South East Asia.
There are two strains of B.malayi;
1. The nocturnal periodic strain, which is widely distributed in Asia, the microfilariae
being in their highest concentrations between the hours of 10pm and 2am.
2. The sub-periodic strain which is found in Malaysia, Indonesia and the
Philippines where humans exhibit a microfilaraemia all the time with the highest
numbers being detected between noon and 8pm.
Nocturnally periodic Brugian filariasis is primarily a rural disease, being
transmitted by various Anopheles species of mosquitoes and also by Mansonia, a
mosquito that usually bites during the night.
Nocturnally sub-periodic B. malayi is transmitted almost exclusively by Mansonia
species, often, different species than those involved in transmitting the periodic form.
Mansonoa bonneae are important vectors in Malaysia, breeding in swamp forest
and biting by night, although sometimes by day as well.
106
Infection with Echinostoma species is thought to be contracted by injestion of
fresh water snails containing metacercaria. Such as Echinostoma ilocannum which
occurs in the Philippines. The metacercariae infect the large snail Piola luzionica
and in return are eaten raw.
Despite the large numbers of these flukes they are of little medical importance,
the most important being F. buski.
Life cycle and transmission
The life cycles of Fasciola, Fasciolopsis and Echinostoma species are complex,
requiring more than one intermediate host.
Adult worms inhabit the liver or bile ducts of the definitive host (human), where
they lay many eggs, which are deposited into the environmnent in the faeces. They
are immature when passed. If they are passed into water they become mature in 9 to
15 days at the optimum temperature of 22-25°C.
The larvae (miracidia) develop within the eggs and hatch out into water, where
they then penetrate the snail (intermediate host). The miracidia must find a snail
within 8 hours of hatching out of the egg. Each species of fluke favours one particular
intermediate host; F. hepatica invade snails from the genus Lymnaea, the most
important being Lymnaea truncatula and F. buski invade snails of the family
Planorbidae, e.g. Segmentina hemisphaerula.
Within the snails the miracidia mature into sporozoites and then into redia and cercariae.
This development takes 1 to 2 months according to the temperature. (In summer there is
only one redial generation but in cooler weather there are two redial generations.)
The cercariae leave the snail, usually at night: they then swim around for several
hours and then encyst (forming a resistant external wall) on submerged vegetation or
fresh water fish.
If the cysts are ingested by a definitive host, the metacercariae excyst in the
duodenum. They then migrate through the intestinal wall and either reach the liver
tissue and bile ducts via the body cavity or via the lymphatics and the circulation.
They require 2 to 3 months to reach maturity.
The life cycle of the Echinostomes differs by one minor point: the cercariae
encyst wither within the tissues of the same molluscan host in which sporocysts and
rediae develop, or penetrate and encyst in other animals such as amphibians or fish.
Morphology
The morphology of the adult flukes of Fasciola, Fasciolopsis and Echinostoma
species is well documented. They are large leaf-shaped parasites about 2-3 cm long.
There are two suckers, an anterior oral sucker surrounding the mouth and a ventral
sucker (acetabulum) on the ventral surface.
The outer tegument is covered in tiny spines, which face backwards enabling
them to attach themselves along with their suckers to the tissues.
Ova are all thin shelled, ellipsoid, quinone coloured (bile stained) with an operculum
that is often inconspicuous. Although ova of Echinostoma species can usually be
differentiated by size due these flukes beign much smaller in size than F. Buski and F.
hepatica, there is much cross-over in the size of Fasciola and Fasciolopsis species.
59
 Pathogenesis
Light infections due to Fasciola hepatica may be asymptomatic. However, they
may produce hepatic colic with coughing and vomiting; generalised abdominal
rigidity, headache and sweating, irregular fever, diarrhoea and anaemia.
Infections due to Fasciola gigantica occur mainly in cattle raising areas and
cause clinical symptoms similar to those of Fasciola hepatica although human
infections are less common.
The adult flukes of Fasciolopsis buski attach to the intestine, resulting in local
inflammation and ulceration. Heavier infections may subsequently lead to abdominal
pain, malabsorption and persistent diarrhoea, oedema and even intestinal obstruction.
Marked eosinophilia may be seen.
The adult flukes of Echinostoma species attach to the intestine resulting in little
damage to the intestinal mucosa. Light infections are generally asymptomatic and
heavy infections may produce light ulceration, diarrhoea and abdominal pain.
Laboratory diagnosis
Definitive diagnosis is made by observing the ova in faeces; since the flukes are
very prolific any significant infection will be easily picked up. Where identification
cannot be made from the size of the ova, clinical information and the source of
infection may help to provide a diagnosis. Serological techniques are available for
the diagnosis of Fasciola hepatica.
The Intestinal Parasite IQA kit is a comprehensive kit containing stained protozoan
slides and formalin fixed suspensions of helminth ova and larvae and a semi-
interactive CD-rom (The CD-rom is an excellent form of interactive reference
material, allowing you to understand easily the complex subject of parasitology). It
is ideal for teaching, reference and internal quality assessments.
Parasep faecal parasite concentrators are provided enabling concentrations to be carried
out on the helminth suspensions. All reagents and products are supplied to prepare slides.
Quality assessments can then be carried out following the staining procedures as
recommended on the picture reference cards.

Liver Flukes:

Clonorchis sinensis, Opisthorchis species, and Dicrocoelium

dendriticum

Classification. Helminths. Phylum Platyhelminthes. Trematodes.

Figure 30. Wuchereria bancrofti
Diseases. Clonorchiasis (Chinese liver fluke infection), Opisthorchiasis,
Dicrocoeliasis. vector. Thus, in nocturnally periodic forms the microfilaria are present in the pe-
Geographic Distribution ripheral blood circulation at night; during the day they reside in the deep tissues,
China. Taiwan, Japan, Korea, Vietnam (Clonorchis); Thailand and Lao People’s particularly the lungs.
Democratic Republic (O. viverrini); Eastern Europe and the former USSR Morphology
(O. felineus}; Europe, former USSR, northern Africa, northern Asia, areas of the Far The adult worms are white and threadlike. The male measures between 2.5-4cm
East, and the western hemisphere (Dicrocoelium). whereas the female is larger, measuring between 8-10cm.
60 105
lion are infected with filariasis. The species which are primarily responsible for
these human filarial infections are; Wuchereria bancrofti, Brugia malayi and On-
chocerca volvulus.
Wuchereria bancrofti
Introduction
Wuchereria bancrofti is a nematode causing lymphatic filariasis throughout the
tropics and subtropics and is transmitted by the mosquito. There are two strains of
W. bancrofti;
1. The nocturnal periodic strain which is widely distributed in endemic regions i.e.
Africa, India and the Far East and also parts of China, Korea and Japan, the microfilariae
being in their highest concentrations between the hours of 10pm and 2am.
2. The sub-periodic strain which is found in the Pacific region, and has a
microfilaraemia all the time with the highest numbers being detected between noon
and 8pm.
Humans are the only known reservoir host of W. bancrofti. Infection rates in
some communities in East Africa exceed 30% of adults causing revolting swellings
of the legs or genital system, known as elephantiasis in man. The adult worm occurs
in tightly coiled nodular masses in the major lymphatic ducts.
The main vector is Culex quinquefasciatus , a mosquito that is particularly
common in towns and cities, breeding in organically polluted water, resting in houses
and feeding by night on their human occupants. Typical breeding sites include: storm
drains blocked with domestic refuse, accumulations of domestic waste water,
inadequately covered septic tanks and pit latrines.
In rural areas throughout Africa Anopheles gambiae and Anopheles funestus are
involved in transmission. Elsewhere other anopheline mosquito species may transmit
bancroftian filariasis in rural areas, while in Papua New Guinea Mansonia may act
as a vector.
Life cycle
Microfilariae enter the host during a blood meal when the vector, a mosquito,
punctures the skin. The infective larvae enter through the wound and migrate to the
peripheral lymphatics where they grow to mature male and female worms (Fig. 30).
They can live there for several years. After mating, the gravid females release sheathed
microfilariae into the peripheral blood where they can be detected 8-12 months after
the initial infected bite.
The mosquito acquires the infection by ingestion of the microfilaria in the blood
meal. The microfilariae lose their sheath on arrival in the stomach of the mosquito
due to gastric juices. The larvae migrate to the thoracic muscles and develop into
infective larvae over a period of 6-14 days. The larvae then migrate to the mouthparts
of the mosquito, which infects the host during a blood meal.
The blood stages of filariae, mifcrofilariae, vary in the times when they are
present in the peripheral blood, corresponding with the peak biting time of the
104
Location in Host. Biliary passages of the liver.
Morphology
Adult Worms. Adult Clonorchis are flattened and spatulate, measuring 10-25
mm long by 3-5 mm wide. They are hermaphroditic, with a single, rounded ovary
situated anterior to the two extensively branched testes. Adults of O. viverrini and
O. felineus from humans are usually smaller, 7-12 mm long, and their slightly lobed
testes are posterior to the oval or lobed ovary. Dicrocoelium adults measure 5-15
mm long by 1,5-2.5 mm in width, and their paired, slightly lobed testes are
immediately behind the ventral sucker and anterior to the ovary.
Eggs
The ovoid, yellow-brown eggs of Clonorchis have a moderately thick shell and
a seated operculum that results in its prominent «shoulders» appearance; they measure
27-35 µm by 12-19 µm. There is usually a small knob at the abopercular end. These
eggs contain a miracidium when passed in feces. Eggs of O. viverrini and O. felineus
are similar in morphology and size, measuring 19-29 µm long by 12-17 µm in width.
Operculate Dicrocoelium eggs are thick-shelled, have a deep golden brown color,
contain a miracidium, and measure 38-45 µm long by 22-30 µm in width.
Life Cycle
Clonorchis and Opisthorchis have similar life cycles involving appropriate
freshwater snails as the first intermediate hosts and many fish as second
intermediate hosts. Metacercariae encyst under the scales or skin of fish. Infection
is acquired by ingestion of raw or inadequately cooked infected fish. Clonorchis
metacercariae, ingested by a mammalian host, migrate to the bile ducts of the
liver via the common bile duct and mature in 3 to 4 weeks. The life span of adult
Clonorchis may be 20 to 25 years. Cats and dogs serve as animal reservoirs for
human infection. Dicrocoelium has a markedly different life cycle that involves
the use of appropriate terrestrial snails as first intermediate hosts. Large numbers
of cercariae are released from snails and agglomerate within the mucus of
molluscs to form «slime balls» that are deposited on soil or grass. These slime
balls are ingested by ants that serve as the second intermediate host; infection is
acquired by the ingestion of ants harboring metacercariae. Sheep, cattle, deer,
and other herbivores are the usual hosts for this parasite, but human infections
are common in many areas.
Diagnosis. Detection of characteristic eggs in feces.
Diagnostic Problems
Clonorchis eggs sometimes are confused with heterophyid eggs but generally
are somewhat larger and have a prominent seated operculum, whereas heterophyid
eggs usually have an inconspicuous operculum flush with its shell surface. Although
Clonorchis eggs typically have a knob or hooklike protrusion at the abopercular
end, it is often difficult to see or may be absent. Although Opisthorcbis eggs are
quite similar to those of Clonorchis, the shoulders of these eggs are not usually as
prominent. Ingestion of infected livers from herbivorous animals may result in
spurious passage of Dicrocoelium eggs in human feces.
61
 Comments
Clonorchis infections occurring in residents of Hawaii and the west toast of the United
States who have not been to the Orient probably are caused by the eating of imported
pickled fish containing still-viable metacercariae. In Canada, a related liver fluke, Metorchis
conjunctus, has been found to cause clinical illness (abdominal pain, anorexia, and liver
function abnormalities) in people eating raw fish, in particular the while sucker, Catostomus
commersoni. Eggs of this parasite measure approximately 28 µm in length by 16 µm in
width and are morphologically indistinguishable from the eggs of Opisthorchis species.
Clonorchis sinensis
Introduction
Clonorchis sinensis, also known as the Chinese liver fluke is a narrow elongate
liver fluke found in the Far East, mainly Japan, Korea, China, Taiwan and Vietnam.
It belongs to the group of Oriental liver flukes where there are three main species,
which commonly infect man. The other two species are Opisthorchis felineus and
Opisthorchis viverrini. The three species are so similar in their morphology, life
cycles and pathogenicity that they are very rarely discussed as separate species.
All members of this group are parasites of fish-eating mammals, particularly in
Asia and Europe. Man is the definitive hosts and water snails and fish are the
intermediate hosts. Infections can be easily avoided by man not eating raw fish since
this is the only way that infection can be passed on.
Clonorchis sinenesis parasitise the biliary duct in humans who become infected by
eating raw or undercooked fish. Dogs and cats are the most important reservoir hosts.
Life cycle and transmission
The adult flukes are found in the bile ducts and gall bladder where they deposit
eggs. The eggs are passed out into the environment in faeces (Fig. 14).
Entered the gut in the bile. Further development can only take place if they are
eaten by appropriate species of water snails (intermediate hosts), e.g. Bulimus
fuchsianus. Within the snails body the miracidium (which hatched out of the
operculated egg) matures into a sporocyst and then a redia (both are asexual replicative
stages). Within the redia, several cercariae develop with unforked tails. These escape
into the surrounding water when the redia finally bursts. They can live in the water
for 1-2 days waiting to come in contact with a suitable species of fish (over 80
species have been recognised as susceptible hosts), they force their way in through
the scales, lose their tails and encyst as infective metacercariae.
Humans become infected when they eat raw or slightly pickled fish, the
metacercariae excyst in the duodenum and descend the bile ducts. There they develop
into adult flukes within 4 weeks.
From infection of the snail to the formation of the infective metacercariae takes
about 8 weeks.
The ova of Clonorchis sinensis contain fully developed miracidia and possess
prominent opercular shoulders (flask shaped egg) and are operculate. They are bile
62
raw pork. Diarrhoea with or without abdominal pain may last for several weeks.
Eosinophilia and fever occur in most cases. Leucocytosis is common and hyper-
globulinaemia is characteristic. Myocytosis and circum orbital oedema are classic
signs. There can also be central nervous system involvement.
Pathogenicity
The primary pathogenic effect of Trichinella comes from the destruction of the
striated muscle fibres in which it encysts. There can be neurological manifestations
of trichinosis and death may be ascribed to myocarditis, encephalitis or pneumonitis.
Laboratory diagnosis
Diagnosis of trichinosis depends on the clinical signs, such as myalgia, periorbital
oedema, fever and eosinophilia in a patient with a history of eating pork or sausages.
Serological tests are available but may be negative if carried out within 3-4 weeks
post infection. Circulating antibodies to T. spiralis appear from 2-4 weeks after
infection. Redefined diagnostic antigens for their detection are currently being
developed. A simple IFAT employing fragments of larvae, as antigen is a useful
diagnostic tool. Latex tests with extracted larval antigens have also proved valuable
in the acute stage, during which high antibody titers develop.
Muscle biopsy is available with the muscle being digested in pepsin, which frees
the encapsulated larvae or by a simple device whereby the muscle sample is
compressed between 2 glass plates to make it semi-transparent, allowing you to see
any encapsulated larvae using a “trichinoscope” (a simple magnifying system).
The Blood Nematodes
These nematodes are known as filariae and consist a group of nematodes, which
have successfully invaded the blood stream, connective tissue, or serous cavities of
vertebrates. They are long thread –like nematodes.
Many of them are of medical and veterinary importance attacking man and various
domestic animals being transported by various vectors, including mosquitoes. The nematodes
from this order do require intermediate hosts for the completion of their life cycle.
The morphology of these nematodes consist of a cylindroid pharynx with an
anterior muscular portion and a posterior glandular portion; the males have well-
developed alae and spirally coiled tails.
Sexually mature female worms release microfilaria, which are pre-larval stages.
These are released into the bloodstream. Most species are known to be ovoviviparous
and some have “sheathed” microfilaria.
The filarial nematodes which parasitise man consist of Wuchereria bancrofti,
Brugia malayi, Brugia timori and Loa loa, Onchocerca volvulus, Mansonella perstans,
Mansonella streptoserca and Dipetalonema streptocerca.
They inhabit a range of locations within the body; lymph glands, deep connective
tissue, subcutaneous tissues or mesenteries. Invasions of these tissues usually result
in inflammatory reactions, which is a typical symptom of a human filarial infection.
In some cases these result in fleshy deformities known as elephantiasis.
It has been estimated that approximately 1 billion people in tropical and sub-
tropical countries are exposed to the risk of filarial infections and at least 200 mil-
103
 Figure 29. Trichinella spiralis

The female adult worms are ovoviparous and up to 1500 larvae may be released
by a single worm.
Clinical Disease
Symptoms during the intestinal phase may go unnoticed or may be severe. Epi-
demics can result in outbreaks of gastro-enteritis, 2 to 7 days after the ingestion of
102
Figure 14. Clonorchis sinensis
stained and measure 29mm by 16mm. In wet mounts they are transparent and you
can quite easily see their anatomy. There can be up to 6,000 worms present and a
daily egg output of 1,000 eggs per microlitre of bile or 600 per gram of faeces.
63
 The cercariae possess eyespots; the penetration and cystogenous glands are also
well developed.
Pathogenesis
Many millions of people become infected every year but only a minority suffers
from any illness. The pathology is related to the number of parasites present. Light
infections of up to 50 eggs or more are usually asymptomatic. A heavy infection of
500 or more eggs may cause serious illness.
Acute infections may be characterized by fever, diarrhoea, epigastric pain,
enlargement and tenderness of liver and sometimes jaundice. The invasion by these
worms in the gall bladder may cause cholecystitis, due to flukes becoming impacted
in the common bile duct.
Laboratory diagnosis
Definitive diagnosis is made by observing the characteristic ova in faeces
following concentration of the faeces or from duodenal aspirates when there is
complete obstructive jaundice or from the Entero-Test.
Table 4
Table summarising the less common flukes that are known to infect man
Metagonimus yokogawai
Classification. Helminths. Phylum Platyhelminthes. Trematodes.
Diseases. Metagonimiasis.
Geographic Distribution
Metagonimus yokogawai is found in China, Japan, southeast Asia, and the Balkan
states. On a worldwide basis, these intestinal fluke typically have highly localized
geographic distributions.
Location in Host. This intestinal fluke lives in the crypts and lumen of the small
intestine.
64
Trichinella spiralis
Introduction
Trichinella spiralis was first seen by James Paget but was named and described
by his Pofessor, Richard Owen. The family Trichinellidae contains only one single
genus Trichinella and was originally thought only to contain the one species,
Trichinella spiralis, which causes the serious and often fatal disease in man known
as trichinosis (trichinelosis). It is a parasite of carnivorous animals and is especially
common in rats and in swine fed on uncooked garbage and slaughter house scraps,
humans become infected by eating raw pork, with sausages being the most common
cause of infection. It is a cosmopolitan parasite and prevalent in many European
countries with the highest interest being in China.
It is now thought that there are four varieties of this species that exists worldwide;
Trichinella spiralis spiralis – temperate zone – high infectivity for pigs, rats and man.
Trichinella spiralis nelsoni – Tropics – low infectivity for pigs and rats and high
infectivity for lions, hyenas.
Trichinella spiralis nativa – Arctic – low infectivity for pigs, found in polar
bears, resistant to freezing.
Trichinella spiralis pseudospiralis- New Zealand – low infectivity for pigs, rats
and mice.
Trichinella spiralis is a “domestic” parasitic nematode long recognized to cause
a zoonosis transmitted to man by the ingestion of infected pork.
Life cycle
Infection in the definitive hosts is acquired by the hosts eating raw or undercooked
flesh e.g. pork, containing encapsulated larvae. Rats are probably the most highly
infected “natural” hosts and pigs become infected by eating infected pork scraps or
occasionally rats, which inhabit their stalls (Fig. 29). For man sausages are a dangerous
source of the parasite as a small fragment of infected pork, (after mincing), may
become widely distributed among a number of sausages.
Humans become infected by eating raw meat containing encysted larvae. The
cyst becomes digested and releases the larvae, which invade the intestinal mucosa.
They develop and mate in the second day. By the 6th day of infection, the female
adults deposit motile larvae, which are carried by the intestinal lymphatics or
mesenteric venules to other tissues in the body. The very active muscles, such as the
diaphragm, jaws, tongue, larynx and eyes, are invaded and the larvae become
encapsulated by the 21st day following infection. Calcification of the cysts occurring
as early as five months, but usually begins after 6-18 months. The cyst wall is derived
from the host’s muscle fiber and the larvae remain viable for many years with no
further development occurring. When muscles are eaten by the definitive host, sexual
maturation in the intestinal phase, as explained earlier, rapidly occurs.
Morphology
The adult female worm is about 2-3mm long and 90mm in diameter. The male is
smaller measuring 1.2mm long by 60mm in diameter.
101
morphology of the esophagus seen in adult worms is present in these larvae and aids
in the correct diagnosis.
Diagnostic Problems
Because eggs are not usually found in routine fecal examinations, cellulose tape
preparations are the most reliable means for detecting this infection.
Comments
Enterobiasis is a familial and group infection that is more prevalent in children.
It is very common in daycare nurseries and institutional settings A second species,
E. gregorii, has been observed from humans; males can be distinguished from those
of E. vermicularis by having a short spicule (70-80 µm long).
Figure 28. Enterobius vermicularis
100
Morphology. Adult Flukes
M. yokogawai are minute, pyriform-.shaped organisms, measuring 1.0-2.5 mm
long by 0.3-0.7 mm wide. Characteristically they have a fleshy collar at the anterior
end that is provided with spines and partially surrounds the oral sucker.
Eggs
Eggs are small, ovoid, operculate, and yellow-brown, measuring 20-30 µm by
15-17 µm. They contain a miracidium when discharged in feces. With most of the
genera listed above, the sizes of their embryonated eggs overlap considerably,
generally between 20-30 µm in length. M. yokogawai eggs resemble the eggs of
Fasciola and Fasciolopsis in shape but are considerably smaller, measuring 80-115
µm by 58-70 µm. These eggs are thin-shelled, unembryonated when laid, and have
an inconspicuous operculum. Frequently, they have a roughening or slight thickening
of their shell at the abopercular end.
Life Cycles
Intestinal flukes use fresh-water snails and fish as first and second intermediate
hosts, respectively. In Fish, the metacercarial stages typically are encysted under the
scales or in the skin, and humans become infected by eating raw or inadequately
cooked fish. In some instances, marine bivalves such as clams and oysters serve as
intermediate hosts. The prepatent period is usually short, from 1 to 3 weeks, and the
normal life span is up to several months.
Diagnosis. Presence of characteristic eggs in feces.
Diagnostic Problems
Because the egg-laying capacity of M. yokagawai and other small intestinal flukes
is limited, sedimentation concentration procedures may be needed to demonstrate
eggs in light infections. Accurate species identification is often difficult because the
eggs of most of these flukes are similar in size and morphology; without knowledge
of the types of intestinal flukes occurring in the animals in any geographic area a
specific identification is frequently impossible.
Eggs of many of the intestinal flukes may be confused with those of the liver
flukes, Cionorchis sinensis and Opisthorchis species. Usually the eggs of the liver
flukes are somewhat larger (27-35 µm by 12-19 µm), have a seated operculum, and
a knob at the abopercular end.
Comments
The presence of this infection in humans, as well as other infections caused by
related, small intestinal flukes, are generally a result of the lack of host specificity by
the parasites. In a particular geographic region where fish are eaten raw or
inadequately cooked, birds, rodents, dogs, cats, and other mammals often serve as
reservoirs for human infections. In addition to the trematodes described above,
numerous other small flukes of animals in southeast Asia occasionally have been
reported from humans.
Pathology caused by this small intestinal fluke is usually limited to mild
inflammatory changes resulting from the attachment of the parasites to the mucosal
epithelium. Mild, intermittent mucous diarrhea and colicky pain are often associated
65
with these infections, but these symptoms are usually limited to a period of several
months to 1 year, because of the short life span of the adult worms.
Paragonimus westermanni
Introduction
Paragonimus westermanni is a lung fluke found in both humans and animals.
The adults are 12mm long and are found in capsules in the lung. Although they are
hermaphroditic, it is necessary for worms to be present in the cyst for fertilization to
occur. The disease is seen in the Far East, China, South Eat Asia and America.
Life cycle
Humans become infected by ingestion of insufficiently cooked crayfish or crabs
containing metacecariae which excyst in the intestine, penetrate through the wall
into the peritoneal cavity and make their way through the diaphragm and pleura into
the lungs (Fig. 15).
The lung cysts in which the worms most commonly occur usually contain 1-3
flukes. The eggs become freed into the bronchial tubes and pass out with sputum,
but they may also appear in the faeces in large numbers, as a result of being swallowed.
Sporocyst and 2 redia generations occur, giving rise to creeping cercariae. These
penetrate a number of fresh-water crustaceans, in which they encyst invarious sites
such as gills, muscles, heart and liver. Encysted metacercariae are not immediately
infective but have to undergo further maturation.
Mammalian hosts ingest the cysts, which are then ingested in the duodenum and
the freed metacercariae penetrate through the intestinal wall into the body cavity to
reach the pleural cavity in about 4 days and the lungs in about 14 – 20 days. In the
lungs, a fibrous capsule is formed by the host, after about 6 weeks the worms mature
and produce eggs, which characteristically appear in the sputum.
Morphology
The adult worm is an ovoid, reddish brown fluke about 12 mm long.
The eggs are ovoid, brownish yellow, thick shelled and operculated. They measure
80-100mm by 45-65mm and may be confused with the ova of Diphyllobothrium
latum.
Clinical Disease
As the parasites grow in the lung cyst, inflammatory reaction and fever occurs.
The cyst ruptures and a cough develops resulting in an increase in sputum. The
sputum is frequently blood tinged and may contain numerous dark brown eggs and
Charcot-Leyden crystals. Haemoptisis may occur after paroxysms of coughing.
Dyspnoea and bronchitis develop with time. Bronchiectasis may occur and pleural
effusion is sometimes seen. The disease resembles pulmonary tuberculosis. Cerebral
calcification may also occur.
Laboratory Diagnosis
Diagnosis is based on finding the characteristic eggs in brown sputum. The eggs
can also be found in the faeces due to swallowing sputum. A chest x-ray may show
66
Strongyloides larvae may be present in the stool in very small numbers and cul-
ture methods maybe needed to encourage the rhabditiform larvae to develop into
filariform larvae and migrate from the sample.
The Enterotest or string test can be used to recover larvae from duodenal aspirates.
Larvae must be distinguished from hookworm larvae especially if it is an older
sample. Rhabditiform larvae are most commonly seen.
A good concentration technique is essential to increase the chances of seeing
larvae, though they are easily killed making diagnosis more difficult.
Serology
Serological tests are of value in the diagnosis of strongyloidiasis when larvae
cannot be found. An enzyme-linked immunosorbent assay (ELISA) using larva
antigen, is usually employed.
Enterobius vermicularis
Classification. Helminth. Phylum Nematoda, Nematode.
Disease. Enterobiasis (pinworm infection, oxyuriasis).
Geographic Distribution. Worldwide.
Location in Host. Cecum, appendix, colon, and rectum.
Morphology. Adult Worms
Males are 2.5 mm long by 0.1-0.2 mm wide, and have a blunt posterior end and
a single spicule, 100-140 µm long. Females are 8-13 mm long by 0.3-0.5 mm
wide, and have a long pointed tail. In both sexes, there are cephalic inflations, and
the esophagus is divided into three parts – corpus, isthmus, and bulb.
Eggs
Elongate, flattened on one side, with a thick, colorless shell, 50-60 µm by 20-30
µm. Eggs are partially embryonated when laid.
Life Cycle
Females usually emerge from the anus at night and discharge their partially
embryonated eggs on the perianal surface (Fig. 28). Eggs embryonate to the infective
first stage within 4 to 6 hours. Infection usually is by direct transmission of eggs to
mouth by hands or through fomites. Parasites develop in the lower intestinal tract,
and the prepatent period is 3 to 4 weeks. Adults normally live for only a few months.
Diagnosis
Eggs usually are detected in cellulose tape preparations applied to the patients
perianal region in the early morning prior to the patient’s bathing or using the toilet.
Eggs are sometimes found in fecal preparations; however, routine diagnosis by fecal
examination is unreliable because eggs are not introduced into the fecal stream.
Instead these eggs are discharged onto the surface of fecal material as it passes
through the rectum. Not infrequently, adult females are seen around the anus or on
the surface of stool specimens. Rarely, immature larval stages of pinworms, espe-
cially female worms, are found in fecal specimens. These developing larvae lack
cephalic inflations for their first 2 to 3 weeks of development, but the characteristic
99
duce eggs. Adult males are unable to attach themselves to the mucosa, therefore, for
any copulation to take place they must mate in the lumen of the intestine.
3.Thenon-infective rhabditiform larvae develop into infective filariform larvae
while passing down the small intestine. Autoinfection occurs when the larvae reinfect
the host by penetrating the intestinal mucosa or the perianal or perineal skin. The
larvae migrate to the lungs via the circulatory system and then return to the intestine.
From initial infection to maturity usually takes less than 4 weeks.
Morphology
The first stage rhabditiform larvae measure approximately250mm long by 20mm
wide. They have a bulbed oesophagus and a short buccal cavity. In an old specimen,
rhabditiform larvae of S.stercoralis must be differentiated from those of hookworm,
which have a longer buccal cavity. The third stage or filariform larva is approximately
500mm long and has a notched tail compared with that of hookworm, which is
sheathed and has a long slender tail.
Adults are slender and possess and extremely long oesophagus, which in the female
extends 1/3 to1/2 of the body. The anal opening is ventral and the tail is pointed.
Eggs are rarely found in the stool as they hatch in the intestine. They are oval and thin
shelled, resembling those of hookworm but are smaller measuring 50-58mm by 30-34mm.
Clinical disease
Disease associated with infections due to S.stercoralisis varied, ranging from
some patients being totally asymptomatic to the hyperinfection syndrome. There are
3 areas of involvement in Strongyloides infections; skin, lungs and intestine.
1. Initial skin penetration of the filariform larvae usually causes very little
reaction, however with repeated infections the patient may mount a hypersensitive
reaction thus preventing the larvae from completing its life cycle. The term larva
currens is used when there is a rapidly progressing urticarial track.
2. The migration of larvae through the lungs may stimulate an immune response,
which can result in a cough, wheezing and fever.
3. Symptoms associated with intestinal strongyloidiasis may mimic a peptic
ulcer due to ulceration of the intestinal mucosa. In heavy infections the intestinal
mucosa may be severely damaged resulting in malabsorption. There may also be
lower gastrointestinal bleeding. Eosinophilia may be high.
Hyperinfection syndrome
The autoinfective capability of larvae may be responsible for long-term infections,
which persist for many years. The parasite and host reach an equilibrium state where
neither host nor parasite suffers any adverse reactions. If this equilibrium is disturbed
e.g.immunosuppression, the infection proliferates with immense numbers of larvae
migrating to every tissue in the body, especially the lungs. This condition is referred
to as disseminated strongyloidiasis. This results in tissue damage, pneumonitis, brain
damage or respiratory failure. Figure 15. Paragonimus westermani
Laboratory diagnosis. Microscopy
Laboratory diagnosis depends on finding larvae in stool, sputum or duodenal cystic shadows and calcification. Serological tests, in particular, the ELISA method,
aspirates. are useful diagnostic tests.
98 67
 The Schistosomes

Introduction
The Schistosomes are blood trematodes belonging to the Phylum Platyhelmintha.
They differ from other trematodes in that they have separate sexes. The male worms
resemble a rolled leaf where they bear the longer and more slender female in a
ventral canal (the gynaecophoric canal). They require definitive and intermediate
hosts to complete their life cycle. There are 5 species of Schistosomes responsible
for human disease; S. mansoni, S. haematobium and S. japonicum with S. mekongi
and S. intercalatum being less common.
They are the only trematodes that live in the blood stream of warm-blooded
hosts. The blood stream is rich in glucose, and amino acids, so along with the plasma
and blood cells, it represents an environment, which is suitable for egg producing
trematodes.
Over 200 million people are infected over at least 75 countries with 500 million
or more people exposed to infection. With the disease spreading due to improved
water supplies being created therefore, forming potentially new habits for snails.
The disease caused is called schistosomiasis or Bilharzia and is the most important
of helminthic diseases.
Infection by the three most common species is the same in both sexes and in all
age groups. Though, S. mansoni and S. haematobium is seen to occur more often
and most heavily in teenagers especially males.
Life Cycle
Adult worms of S. mansoni live in the plexus of veins draining the rectum and
colon, and in branches of the portal vein in the liver.
Adults of S. japonicum live in the anterior mesenteric blood vessels and in the
portal vein in the liver. Whilst the adults of S. haematobium live in the vesical,
plexus draining the bladder.
Once the eggs are laid by the adult female worms the majority of them first pass
through the veins of the blood vessel in which the worm is living, and then into the
lumen of the intestine and are passed in the faeces (S. mansoni and S. japonicum).
Or into the lumen of the bladder, and are then passed in the urine (S. haematobium).
Those eggs that reach fresh water hatch, releasing a miracidium which, to develop
further must infect a snail of the correct species within 24 hours. The eggs of each
species are markedly different but each produce virtually identical miracidium.
Asexual multiplication takes place in the snail, and results in the release of
cercariae (minute in size with forked tails, 200mm long) into the water about 3-6
weeks later. Cercariae actively swim around and when they have located, or come Figure 27. Strongyloides stercoralis
into contact with, a definitive host they actively penetrate the skin. They can stay
active looking for a host for 24-48 hours after which if they don’t find a host they The larvae, which infect the host by penetrating the skin, undergo a migration
will die. The head of the cercariae migrates to the liver and develops into either adult through the dermal tissues and into the circulation to the heart and lungs, then up the
male or female worms (flukes), here they pair up and then migrate to their region of bronchi and trachea, where they are eventually swallowed and pass down into the
the venous blood system (species specific sites). The females leave the males and intestine. On reaching the mucosa of the duodenum the females develop and pro-
68 97
 Symptomatic infections are usually only seen in children. The majority of infec-
tions are chronic and mild, with nonspecific symptoms like diarrhoea, anaemia, growth
retardation, and eosinophilia.
Laboratory Diagnosis
The adult worms of T. trichiura are rarely seen in the faeces. The microscopic
examination of stool deposits after concentration reveals the characteristic barrel
shaped ova. In symptomatic infections numerous numbers of eggs can be seen due
to the prolific nature of the female worms, even in light infections many eggs can be
seen in the smear.
Strongyloides stercoralis
Introduction
Strongyloides stercoralis is an intestinal nematode commonly found in warm
areas, although it is known to survive in the sub-tropics (hot and humid conditions).
The geographic range of Strongyloides infections tend to overlap with that of
Hookworm due to the eggs requiring the same environmental conditions to induce
embryonation.
This parasite is interesting in that it contains a free-living stage (exogenous) and
a parasitic stage (endogenous) where they larvae undergo development in both stages.
Life cycle
The life cycle of S. stercoralis is a complex one as demonstrated in the figure
below (Fig. 27).
The life cycle has three phases:
The parasitic adult females lay eggs while they are in the duodenum where they
hatchproducing rhabditiform (non-infective) larvae.
1.Thelarvae can have two fates in life, one where they are passed out in the
faeces to continue down the free-living pathor they develop into infective
filariform larvae whilst travelling down thesmall intestine.
2.Thelarvae which, develop in the environment can also undergo different
development. Some larvae undergo directdevelopment (homogonic) or indirect
development (heterogonic).
The non-infective first stage (rhabditiform) larvae develop into free living adults
in the soil within 2-5 days andproduce infective third stage or filariform larvae which
can penetrateexposed skin (heterogonic development). This phase is common in
moist,warm tropical countries.
The non-infective rhabditiform larvae, which are excreted in the faeces, develop
into infective filariform larvae in the soil (homogonic development). These infective
larvae penetrate exposed skin. There is no development of free living adult worms
and this phase is common in temperate zones. The larvae never undergo sexual
maturity.
Both types of larvae can become established in the host by penetrating the skin
or by oral ingestion.
96
moves to smaller venules closer to the lumen of the intestine or bladder to lay her
eggs (about 6 weeks after infection). The majority of adult worms live from 2-4
years, but some can live considerably longer.
Schistosoma mansoni
Introduction
S. mansoni occurs in West and Central Africa, Egypt, Malagasy, the Arabian
Peninsula, Brazil, Surinam, Venezuela and the West Indies. The intermediate host is
an aquatic snail of the geuns Biomphalaria. Man is the most common definitive
host; occasionally baboons and rats are infected.
The adult worms live in smaller branches of the inferior mesenteric vein in the
lower colon.
Morphology
The adult males measure up to 15 millimetres in length and females up to 10
mm. The schistosomes remain in copula throughout their life span, the uxorious
male surrounding the female with his gynaecophoric canal. The male is actually flat
but the sides roll up forming the groove. The cuticle of the male is covered with
minute papillae. The female only posses these at the anterior and posterior end as
the middle section being covered by the male body. Oral and ventral suckers are
present, with the ventral one being lager serving to hold the worms in place, preventing
them being carried away by the circulatory current (Fig. 16).
The ova of S. mansoni are 114-175mm long by 45-68mm wide. They are light
yellowish brown, elongate and possess a lateral spine. The shell is acid fast when
stained with modified Ziehl-Neelsen Stain.
A non-viable egg is dark coloured and shows no internal structural detail or
flame cell movement. Eggs can become calcified after treatment and are usually
smaller, appear black and often distorted with a less distinct spine.
The schistosomes differ from other trematodes in that they are dioecious, digenetic,
their eggs are not operculate and infection is acquired by penetration of cercaria
through the skin.
Clinical Disease
The clinical disease is related to the stage of infection, previous host exposure,
worm burden and host response. Cercarial dermatitis (swimmers itch) follows skin
penetration and results in a maculopapular rash, which may last 36 hours or more.
After mating, the mature flukes migrate to the venules draining the large intestine.
Their eggs are laid and they penetrate the intestinal wall. They are then excreted in
the faeces, often accompanied by blood and mucus.
It is the eggs and not the adult worms, which are responsible for the pathology,
associated with S. mansoni infections. The adult flukes acquire host antigen, which
protects them from the host’s immune response.
The host’s reaction to the eggs, which are lodged in the intestinal mucosa, leads
to the formation of granulomata and ulceration of the intestinal wall. Some of the
69
 Figure 16. Schistosoma mansoni

eggs reach the liver via the portal vein. The granulomatous response to these eggs
can result in the enlargement of the liver with fibrosis, ultimately leading to portal Figure 26. Trichuris trichiura
hypertension and ascites. The spleen may also become enlarged. Other complications
may arise as a result of deposition of the eggs in other organs e.g. lungs. caecum. Abdominal cramps, tenesmus, dysentery and prolapsed rectum may occur
Katayama fever is associated with heavy primary infection and egg production. in these cases.
Clinical features include high fever, hepatosplenomegaly, lymphadenopathy, If a prolapsed rectum is observed, many worms may be seen adhering to the
eosinophilia and dysentery. This syndrome occurs a few weeks after primary infection. mucosa of the rectum.
70 95
the eggs requiring the same conditions to allow for embryonation both species can
be found in human together.
There are several species within this genus each infecting specific hosts, but only
T. trichiura infects man. Causing human trichuriasis. It is a parasite that infects many
more people than is generally appreciated, up to 800 million people throughout the
tropics and temperate regions.
Life cycle
Eggs require a warm, moist environment with plenty of oxygen to ensure
embryonation, but once they have embryonated they are extremely resistant to
environmental conditions.
Adult worms are found in the caecum and upper part of the colon of man. In
heavy infection they can be found in the colon and the terminal ileum. They attach to
the mucosa by the anterior end or by embedding the anterior portion of the body in
the superficial tissues. Obtaining nutrition from the host tissues (Fig. 26).
Once fertilised the female worms lay several thousands of eggs, which are
unsegmented at the oviposition and are passed out in the faeces. Once they have
been passed out they require an embryonation period in the soil, which may last
from 2 weeks to several months, after which they become infective.
When embryonated eggs are swallowed by human hosts larvae are released into
the upper duodenum. They then attach themselves to the villi lower down the small
intestine or invade the intestinal walls. After a few days the juveniles migrate slowly
down towards the caecum attaching themselves to the mucosa, reaching their final
attachment site simultaneously.
The larvae reach maturity within 3 weeks to a month after infection, during which
they undergo 4 moults. There is no lung migration and the time from ingestion of
infective eggs to the development of adult worms is about 3 months.
Infection is achieved by swallowing soil that contains embryonated eggs.
Therefore, children are most commonly seen to possess the infections, as they are
more likely to swallow soil.
Morphology
The adult worms of T. trichuria are characterised by the enormously elongated
capillary-like oesophagus (anterior end). With the anus situated in the extreme tip.
The thin anterior portion of the worm is found embedded in the mucosa. There
are no lips and the vulva is at the junction of the thread-like and thickened regions of
the body. The posterior end is much thicker and lies free in the lumen of the large
intestine.
The female measures 35-50mm long and the male 30-45 mm long.
The ova are characteristically barrel shaped, bile stained with bipolar plugs. They
measure 50-54mm by 20-23mm.
Clinical Disease
Most infections due to this nematode are light to moderate with minimal or no
symptoms. However, a heavy worm burden may result in mechanical damage to the
intestinal mucosa due to the adult worm being threaded into the epithelium of the
94
Laboratory Diagnosis. Microscopy
Laboratory confirmation of S. mansoni infection can be made by finding the
eggs in the faeces. When eggs cannot be found in the faeces a rectal biopsy can be
examined.
Serology
Serological tests are of value in the diagnosis of schistosomiasis when eggs cannot
be found. An enzyme linked immunosorbent assay (ELISA) using soluble egg antigen,
is employed at HTD.
Schistosoma japonicum
Introduction
Schistosoma japonicum is found in China, Japan, the Philippines and
Indonesia. It causes disease of the bowel with the eggs being passed out in the
faeces (Fig. 17).
It differs form S. mansoni and S. haematobium in that it is a zoonosis in which a
large number of mammals serve as reservoir hosts, cats, dogs and cattle playing
major roles in the transmission of the disease.
The life cycle is not very different from that of S. mansoni, the intermediate hosts
are from the subspecies Oncomelania hupensis. Sexual maturity is reach in about 4
weeks and eggs may be seen in the faeces as quickly as 5 weeks.
They worms live coupled together in the superior, mesenteric veins and desposit
1500-3500 eggs per day in the vessels of the intestinal wall. The eggs infiltrate
through the tissues and are passed in the faeces.
Morphology
The adult worms are longer and narrower than the S. mansoni worms. The ova
are about 55-85mm by 40-60mm, oval with a minute lateral spine or knob.
Clinical Disease
The main lesions are again due to the eggs, occurring in the intestine and liver.
The eggs which are sequesters in the intestine mucosa or submucsa initiate
granulomatous reactions, resulting in the formation of pseudotubercules
Due to the number of eggs released by the females the infection is more severe
than one with S. mansoni. This is also due to the parasite being less well adapted to
man, therefore, the circumoval granuloma is very large. The initial illness can be
prolonged and sometimes fatal.
Laboratory diagnosis. Microscopy
Laboratory confirmation of S. japonicum infection can be made by finding the
eggs in the faeces. When eggs cannot be found in the faeces a rectal biopsy can be
examined.
Other Intestinal Schistosome species
Other Schistosome species, which are responsible for human disease, are S.
mekongi and S. intercalatum. These 2 species cause similar symptoms to that of S.
mansoni and can be summarised in table.
71
 Chronic infections may lead to iron deficiency and anaemia resulting from the
excessive loss of iron. Heavy worm burden in children may have serious conse-
quences including death.
Cutaneous larva migrans
If man comes in contact with hookworm larva of the dog (or cat), A. braziliense
or A. caninum, penetration of the skin may take place. The larvae are unable to
complete the migration to the small intestine and become trapped. Trapped larvae
may survive for weeks or even months, migrating through the subcutaneous tissues.
Trapped larvae have been known to produce severe reaction, forming tunnels
through the tissues, causing intense itchy skin eruption, producing a red, track under
the skin, which demonstrates accurately the wanderings of the larvae.
Often intense pruritis and scratching may lead to secondary bacterial invasion,
known as “creeping eruption” or “cutaneous larval migrans”.
First-stage rhabditoid larvae that hatch from eggs are 250-300 µm long by
17 µm. They have a long buccal canal and their genital primordium is small and
difficult to see. Infective, third-stage, filariform larvae are 500-600 µm long. These
have a pointed tail and a ratio of esophageal to intestinal length of 1:4. The sheath
about the larvae is conspicuously striated.
Laboratory Diagnosis
Adults of Hookworm species may be passed out spontaneously in faeces. The
microscopic examination of stool deposits after concentration reveals the
characteristic ova.
Diagnostic Problems
Eggs of this species are indistinguishable from those of Ancylostoma duodenale.
If these eggs hatch in feces because of a delay in fecal examination, the first-stage
larvae must be differentiated from those of Strongyloides stercoralis, which normally
are passed in feces. Whereas hookworm first-stage larvae have a long buccal canal
and an inconspicuous genital primordium, the larvae of Strongyloides have a short
buccal canal and a prominent genital primordium. Stool specimens must not be
refrigerated before attempting to culture larval stages, as Necator is especially
sensitive to cold.
Comments
Because hookworm species cannot be differentiated on the basis of their eggs, it
is necessary to culture larvae or to recover adult worms for morphologic study to
make a specific diagnosis.

Trichuris trichiura

Introduction
Trichuris trichiura, more commonly known as the “whip worm”, due to the
whip-like form of the body. They have a cosmopolitan distribution, though; it is
more commonly seen in tropical climates and in areas where sanitation is poor. They
Figure 17. Schistosoma japonicum seem to occur in areas particularly where Ascaris and Hookworm are found due to
72 93
deposit their eggs whilst in the gut (they can produce up to 20,000 eggs per day); the
eggs are then passed out in the faeces. The rhabditiform larvae hatch in warm, damp
soil (light sandy loam), feeding on bacteria. After about one week during which they
have gone through 2 moults become infective and climb into a suitable position
waiting for a suitable host to pass by. The larvae enter the host by penetrating unbroken
skin (it is now recognised that A. duodenale can successfully enter man by oral
ingestion, this may be more important for this species than skin penetration). The
larvae then enter blood vessels and are carried to the heart, lungs and trachea. They
are then swallowed and develop into adult worms in the small intestine. Larvae that
are initially swallowed may not show this migration.
Larvae live for an average of 3-6 weeks in the tropics (A. Duodenale can live at
lower temperatures than N. americanus can, and so is found in more temperate
climates).
Morphology
Both species have similar general morphology and measure approximately,
females 10-13mm and males 8-11mm. The general morphology of the two species
resembles those of Nippostrongylus brasiliensis, the rat hookworm, but they are
approximately twice the size of the rat hookworm (species not discussed here).
The male species has a posterior copulate bursa, which is absent from the female.
The females though possess a vulva opening, which is found almost one third of the
body length from the posterior end, they also have two ovaries. Most of the female
body is occupied with eggs.
The mouth (or buccal cavity) of the two species show a conspicuous pair of
chitinous plates on the dorsal surface. Ancylostoma duodenale buccal cavity bears 2
hook like teeth on the top and 2 triangular cutting plates on the bottom. While the
mouth of N. americanus has 4 cutting plates, 2 on the ventral and 2 on the dorsal
surfaces. The head is curved in both species but Necator adults it is finer but more
pronounced forming a definite “hook” at the anterior end. The buccal cavity is used
to attach the worms securely to the mucosa of the small intestine. With the teeth and
cutting plates used to pierce the mucosa.
The bursa (the characteristic external feature which forms an umbrella-like
extension surrounding the cloaca) of both male species is well developed, Necator
adults are distinguished from Ancylostoma by the split dorsal rays and the close
arrangement of the lateral rays.
The ova are oval and transparent with a smooth thin shell and measure 56-75mm
by 36-40mm. They are usually passed in the 4-8 cell stage in faeces and may be
embryonated. The ova of both species of Hookworm are similar.
Clinical Disease
Larval penetration of the skin may lead to pruritis, often termed as ‘ground itch’
at the site of penetration. Respiratory symptoms may arise during the larval migration.
The adult worm in the intestine may cause intestinal necrosis and blood loss as a
result of the attachment of the adult to the intestinal mucosa. Patients with acute infec-
tions may experience nausea, vomiting, abdominal pain, diarrhoea and eosinophilia.
92
Table 5
Table describing the other less common intestinal Schistosome species
that are known to cause disease in man
Schistosoma haematobium
Introduction
Schistosoma haematobium is different from the other two species previously
mentioned in that it causes urinary schistosomiasis. It occurs in Africa, India and the
Middle East. The intermediate host is the Bulinus snail.
Just like S. mansoni, its distribution runs parallel to the irrigation projects and in
areas, which favour the intermediate hosts. They are exclusively parasites of man.
The mature worms live in copula mainly in the inferior mesenteric veins and the
females deposit their eggs in the walls of the bladder and finally making their way
into the urine. The life cycle is very similar to that of S. mansoni, with sexual maturity
being reached within 4-5 weeks, but eggs may not appear in the urine until 10-12
weeks or even later (Fig. 18).
Morphology
The adult worms are longer than those of S. mansoni. The ova are relatively
large, measuring 110mm-170mm in length and 40mm-70mm in width. They have an
elongated ellipsoid shape with a prominent terminal spine.
Clinical Disease
The clinical disease is related to the stage of infection, previous host exposure,
worm burden and host response. Cercarial dermatitis (Swimmer’s Itch) following
skin penetration results in a maculo-papular rash and can last 36 hours or more. The
mature flukes of S. haematobium migrate to the veins surrounding the bladder. After
mating, the eggs are laid in the venules of the bladder and many penetrate through
the mucosa, enter the lumen of the bladder and are excreted in the urine accompanied
by blood. Thus haematuria and proteinuria are characteristic, though not invariable
features of urinary schistosomiasis.
As with all Schistosoma species, it is the eggs and not the adult worms which are
responsible for the pathology associated with S. haematobium. In chronic disease,
eggs become trapped in the bladder wall resulting in the formation of granulomata.
Following prolonged infection, the ureters may become obstructed and the bladder
becomes thickened resulting in abnormal bladder function, urinary infection and
73
 Figure 25. Necator americanus

Ancylostoma duodenale is an Old World hookworm; it is the only species of

Europe and areas bordering the Mediterranean. It can also be found in the Middle
East, North China, Africa, Asia and South America.
Life cycle
The adult worms live in the small intestine, attached firmly to the mucous mem-
Figure 18. Schistosoma haematobium brane of the gut lining, and feed on blood and tissue (Fig. 25). The adult females
74 91
are easily recognised, oval in shape with a thick wall showing an irregular bumpy sur-
face. They measure 45-75mm by 35-50mm. The outer covering has an albuminoid coat,
stained golden brown by bile. The outer wall lies directly on top of a thick smooth shell,
which is not easily distinguishable. Some have lost their albuminoid wall. The unfertilised
ova are longer and narrower than the fertile ova, measuring 75-85mm by 35-50mm.
The shell layers of the egg provide a very resistant structure, which can withstand
many chemicals, which make them ideal parasites of the intestine (Fig. 24).
Clinical Disease
Small burdens of worms in the intestine may cause no symptoms. The patient may
have symptoms of pneumonitis with cough and low-grade fever during the migration
of the larvae through the liver and lungs. This can be accompanied by wheezing,
coughing and eosinophilia. In heavy worm burdens the adult worms actively migrate
in the intestine resulting in intestinal blockage, vomiting and abdominal pain but
infections may also be asymptomatic. The worms can penetrate through the wall of the
intestine, or into the appendix, travel up the common bile duct, which may become
blocked or they may then enter the gal bladder or liver. A heavy worm burden in
children may lead to severe nutritional impairment and retardation in growth.
Laboratory diagnosis
The adults of A. lumbricoides may be expelled through the anus, mouth or nose.
It is important to distinguish the adult worms from earthworms, which are segmented
and are often collected as a contaminant from toilets.
The microscopic examination of stool deposits after concentration (refer to volume
1) reveals the characteristic bile stained ova. Eggs may be difficult to identify if an
excess of iodine is added to the wet preparation as they retain the stain thus resembling
debris. Ova may also become decorticated. In most symptomatic cases identification
is easy due to the vast number of eggs, which can be found within a few seconds of
starting to scan the slide.
Hookworm species
Introduction
Hookworms infective to man comprise of 2 species, Necator americanus and
Ancylostoma duodenale. They are classed as one of the most destructive of human parasitic
helminths. There is no intermediate host, with man being the only definitive host.
It is estimated that there are some 900 million cases of infection world wide
(Crompton, 1989). The infection is serious where the worms insidiously undermine
the health of their hosts.
They occur in areas where sanitary and environmental conditions favour the
development of the eggs and larval infections (warm, damp soil).
The geographic distributions of the two species are remarkably divided into;
N. Americanus which predominately is a New World hookworm, where it was
introduced from Africa to the Western Hemisphere. It can also be found in the Far
East, Asia, Africa, South America and Oceania.
90
kidney damage. Chronic urinary schistosomiasis is associated with squamous cell
bladder cancer. Heavy infections in males may involve the penis resulting in scrotal
lymphatics being blocked by the eggs.
Laboratory diagnosis
The definitive diagnosis of urinary schistosomiasis is made by finding the
characteristic ova of S. haematobium in urine. Terminal urine should be collected as
the terminal drops contain a large proportion of the eggs. The urine can either be
centrifuged and the deposit examined microscopically for ova. Eggs can sometimes
be found in seminal fluid in males.
A bladder biopsy is seldom necessary to make the diagnosis. A rectal snip may
show the presence of ova as they sometimes pass into the rectal mucosa.
Serological tests can be of value when eggs cannot be found in clinical samples.
An enzyme linked immunosorbent assay using soluble egg antigen to detect
antischistosome antibody is most sensitive.
There is a marked periodicity associated with the time when most eggs are passed
out. Higher numbers of eggs are encountered in urine specimens passed between 10
am and 2pm, presumably as a result of changes in the host’s metabolic and physical
activities.
TOPIC: PLATHELMINTHES, CESTOIDEA
Larval Cestodes, which Infect Man
Infections in man with Echinococcus granulosus, Echinococcus multilocularis
and Multiceps multiceps are caused by the accidental ingestion of eggs, which are
excreted by the definitive animal host. The disease that is produced due to the invasion
of these parasites is caused by the larval stages or hydatid cyst, is known as hydatid
disease or hydatidosis.
Each cestode possesses an elongated tape-like body, which lacks an alimentary
canal. The adult tapeworms are strings of individuals having a complete set of
reproductive organs (proglottids) in progressive degrees of sexual maturity and
budding off from a body attached to the host tissue by a head or scolex.
The larval stage, show a wide variation being found in almost any organ of both
vertebrate and invertebrate hosts.
Echinococcus granulosus
Introduction
Echinococcosis or Hydatid disease in man is caused by the larval stage of the
dog tapeworm, Echinococcus granulosus. Hydatid disease is most extensively found
in East Africa, North Africa, South Africa, the Middle East and parts of South America
and Australia. The intermediate hosts are cattle, sheep, pigs, goats or camels and the
definitive host for this disease is the dog or other canids.
75
 Life cycle of the cestode
Larval infection in man causes hydatid disease.
Adult worms are only seen in the definitive hosts, dogs, they cannot develop in
man (Fig. 19). Man is an accidental intermediate host of hydatid disease. When the
ova are ingested by a suitable intermediate host, they hatch in the duodenum and the
oncosphere migrates to the blood stream where it is carried to the liver, lungs and
other organs of the body. Here it develops into a hydatid cyst, which consists of an
outer thick laminated cyst wall, and an inner, thin nucleated germinal layer. From the
inner layer brood capsules are produced which contain protoscoleces. The brood
capsules detach from the germinal layer, releasing free protoscoleces. Hydatid sand
is the name given to the fluid in the cysts, which consists of protoscoleces, tissue
debris, and sometimes free hooklets. Here, the life cycle stops in humans, but is
continued when a hydatid cyst containing protoscoleces egg in sheep liver, is ingested
by a suitable canine host where the protoscoleces develop into adult worms.
Morphology
The adult worm measures approximately 3-8.5mm long. The scolex has 4 suckers
and a rostellum with hooks, the latter becoming tightly inserted into the crypts of
Lieberkühn. The mature strobila has only 3-4 proglottids, one is immature, one is
mature and the final one is gravid; when gravid the eggs are expelled in the faeces.
Due to the close similarity of the eggs to other Taenia species found in dogs they
were until recently thought to be morphologically indistinguishable.
The larvae in man develop into a unilocular cyst which gives rise to unilocular
hydatid disease. This is characterised as having only one bladder or many completely
isolated bladders, each enclosed in its own well-developed envelope. The latter consists
of several layers, the most prominent being the laminated layer. Within this again is the
germinal membrane from which the brood capsules arise inside which develop
thousands of larvae or protoscoleces, the whole being suspended in a hydatid fluid.
Clinical disease
Hydatid disease in humans is potentially dangerous depending on the location of
the cyst. Some cysts may remain undetected for many years until they become large
enough to affect other organs. Symptoms are then of a space occupying lesion. Lung
cysts are usually asymptomatic until there is a cough, shortness of breath or chest
pain. Hepatic cysts result in pressure on the major bile ducts or blood vessels.
Expanding hydatid cysts cause necrosis of the surrounding tissue.
Slow leakage of the hydatid fluid results in eosinophilia and rupture of an
abdominal hydatid cyst results in severe allergic symptoms.
Symptoms may not manifest themselves for 5-20 years after the infection. Figure 24. Ascaris lumbricoides
Laboratory Diagnosis
1. Imaging and serodiagnosis are the mainstay of diagnosis. Serological tests spicules. The female is 20-35cm long with a diameter of 5mm with a straight pointed
include Enzyme linked immunosorbent assay (ELISA), an indirect haemagglutination posterior end. The mouth has one dorsal and 2 ventral lips. Both are creamy white
test a complement fixation test and a Western Blot system. and the cuticle has fine circular striations.
2. Microscopic examination of the cyst fluid to look for the characteristic The ova can be unfertilised, fertilised or decorticated and can show considerable
protoscoleces, which can be either invaginated or evaginated. The cyst fluid will variation in shape and size. They measure 85-95mm by 43-47mm. The fertilised ova
76 89
the vitamin by the adult worm and the absorption of cobalamins from the host intestine
(occurring only in a small percentage of people).
Laboratory diagnosis
Laboratory diagnosis depends on the recovery of characteristic eggs from a formol
ether concentrate of faeces. Proglottids may also be seen in faecal samples usually
in a chain of segments from a few centimeters to about 0.5 meters in length.

TOPIC: NEMATHELMINTHES, NEMATODA

The Nematodes

Nematodes (or ‘round worms’) are non-segmented helminths known as make up a

large assemblage of relatively simple structured organisms. They possess bilateral symmetry
and a complete digestive tract with oral and anal openings; they taper to a relative point at
both ends. They are also found to have separate sexes, with the male being smaller than the
female, ranging in size from a few millimeters to over a meter in length. Their cylindrical
non-segmented bodies allow them to be easily distinguishable from other helminths.
Nematode infections have a wide spread distribution being found in both
Temperate and Tropical climates. They can be found in fresh water, in the sea and
the soil, successfully invading both animals and plants. The nematodes found in
man invade the body fluids such as the blood or lymph channels and also the
intestine. The ones that successfully invade the intestine are generally larger but,
the nematodes, which invade the tissues, can grow to relatively enormous lengths.
Once hatched in the intestine they undergo an incredible migration. The larvae
initially burrow into the mucosa, penetrate blood vessels and appear as second stage
larvae in the liver within six hours post-infection. Here they remain for several days
and develop into third stage larvae, L3. These larvae then migrate to the heart and are
carried to the lungs via the pulmonary arteries, arriving within 4 to 7 days. From there
they break out of the capillaries into the alveoli and finally work their way up the
trachea to the pharynx and reach the small intestine on the 8th or 10th day post-infection.
Within the intestine, the larvae begin their third moult and become fourth stage
larvae by the tenth day. The pre-patent period of A. suum in pigs (40-53 days) is less
than that of A. lumbricoides (54-61 days) in humans. Two to three months after
ingestion of the eggs, the females lay eggs in the intestine.
The fertilised female can lay about 200,000 eggs per day. Eggs require oxygen
and moisture to embryonate and the worm is often found associated with Trichuris
trichiura (see the Trichuris trichiura section).
Figure 19. Echinococcus granulosus
Ascaris lumbricoides

Morphology also reveal free hooklets and tissue debris. 1% eosin may be added to the fluid to
Ascaris lumbricoides is the largest of the intestinal nematodes found in man. The determine the viability of the protoscoleces. Viable protoscoleces exclude eosin
male measures 15cm with a diameter of 3-4mm and has a curled tail with protruding whereas nonviable protoscoleces take up the eosin.
88 77
 3. Histological examination of the cyst wall after surgical removal.
Western Blots
One serological test, which has proved to be of value to diagnosing Hydatid
disease, is the Western Blot. The test presents a definitive means for detection of
human antibodies to the cestode E. granulosus.
Diagnosis can be achieved using the Western Blot assay for the detection of IgG
antibodies in serum reactive with E. granulosus antigens present on a membrane.
Field studies support a sensitivity of 80% and specificity of 100% in patients with
hepatic cysts.
This assay is known as the Qualicode Hydatid Disease Kit, the principle behind
the test is that it is a qualitative membrane-based immunoassay manufactured from E.
granulosus proteins. The E. granulosus proteins are fractionated according to molecular
weight by electrophoresis on a ployacrylamide slab gel (PAGE) in the presence of
sodium dodecyl sulfate (SDS). The separated E. granulosus proteins are then transferred
via electrophoretic blotting from the gel into strips for testing of individual samples.
During the procedure, the strips containing the E. granulosus proteins are
incubated with serum specimens and washed to remove unbound antibodies.
Visualisation of human immunoglobulins specifically bound to E. granulosus
proteins is performed by sequential reaction with goat anti-human immunoglobulin-
alkaline phosphatase conjugate and BCIP/NBT substrate. Bands corresponding to
the positions of the resoled E. granulosus proteins will be visualized on the strip,
indications the presence in the serum sample of IgG antibodies direct against E.
granulosus antigens. Band positions are compared to those on a reference strip
developed using the Hydatid disease positive control.
Prevention
1. Safe disposal of dog faeces.
2. Education to prevent feeding uncooked offal to dogs.

Echinococcus multilocularis

Introduction
The larvae of Echinococcus multilocularis is a particularly dangerous species
causing multilocular (alveolar) hydatid disease in man and animals and is common
in the highlands of Europe i.e. Switzerland and Germany, in Canada, Alaska and
Northern Russia. The most common definitive hosts are foxes and wolves in addition
to domestic cats and dogs when they have access to infected rodents. Figure 23. Diphyllobothrium latum
Life cycle
Foxes are the primary definitive hosts although in domestic circumstances dogs Clinical Disease
can act as the definitive host. Rodents are the intermediate hosts. Man is an accidental The infection caused by D. latum is due to the ingestion of raw, poorly cooked or
host by the ingestion of eggs where multilocular cysts are formed. In these cysts, the pickled fresh water fish. The symptoms associated with D. latum infection may be
limiting membrane is thin and the germinal epithelium may bud off externally resulting absent or minimal with eosinophilia. There may be occasional intestinal obstruction,
in proliferation in any direction. Metastases may occur. Unlike E. granulosus, there diarrhoea, and abdominal pain. The most serious symptom is the onset of pernicious
is little fluid in the cysts of E. multilocularis. anaemia. This is due to a vitamin B12 deficiency, caused by excessive absorption of
78 87
quality control and training purposes. Intersep can supply you with the most important
fixatives and reagents that are used in clinical laboratories. Accompanying these fixatives,
there is a range of permanent and temporary stains available to suit your needs.
All stains, reagents and fixatives are supplied prediluted ready for use.
Diagnosis is based on recovery and identification of the characteristic ova in a formol-
ether concentrate of faeces. Adult worms and proglottids are rarely seen in stool samples.
Diphyllobothrium latum
Introduction
Members of this order, commonly known as pseudophyllids, are chiefly parasites
of fish-eating mammals, birds and fish. They typically are found with a scolex which
is characterised by two shallow elongated bothria situated with one dorsally and one
ventrally. The proglottids are flattened dorsoventrally.
Diphyllobothrium latum is an intestinal tapeworm, known as the human ‘broad’
tapeworm. It is the largest tapeworm found in man. The term ‘broad’ relates to the
fact that the proglottids are generally wider than they are long. It is an important
human parasite. The adult worms of two other species of the genus, D. dendriticum
and D. ditremum are chiefly parasite of fish-eating birds and mammals.
The tapeworm, D. latum has a wide distribution, occurring especially in countries
bordering the Baltic Sea (Finland, Sweden etc.): and also in Russia, Switzerland and
North America. It is in these countries where the populations are known to eat
uncooked or partly cooked (i.e. smoked) fish.
Apart from man they are found in many other hosts, especially the dog, cat and
pig. This is due to the host countries allowing the domestic animals access to the
offal from the infected fish.
Life cycle and transmission
The life cycle of this tapeworm requires two intermediate hosts (Fig. 23).
The eggs are passed out in human faeces, once in water they hatch out into small
ciliates coracidium larvaem, which swim until ingested by Copepods. It is in these
intermediate hosts that growth and development of the 1st larval stage are completed
(They are now known as procercoids). When these crustaceans (fresh water) are eaten
by fish, the procercoid larvae continue to develop in the flesh of the fish and become
known as plerocercoid larvae. It is this stage of the larvae, which develops in man
when they eat undercooked fish and they grow into adult worms in the small intestine.
Morphology
The egg is usually ovoid and has a small knob at the opercular end and is yellowish-
brown in colour with a smooth shell, of moderate thickness. They measure 58-75mm
by 40-50mm in size.
Adult worms can reach up to a length of 10 metres or more and may contain up to
3,000 proglottids. The scolex is spatulate with no rostellum or hooklets. It has 2 shallow
grooves or bothria, which are unlike the typical 4 suckers seen on the Taenia species. The
proglottids measure 3mm long and 11mm wide and have a rosette shaped central uterus.
86
Morphology
The morphology is in general very similar to that of E. granulosus, but the
adults are much smaller.
Unlike E. granulosus, cysts of E. multilocularis in man do not contain daughter
cysts with scolices. Instead the larval cyst, or as it is referred to as an alveolar or
multilocular hydatid cyst forms a multicystic structure made up of proliferating
vesicles embedded in a dense fibrous stroma, which is often mistaken for a hepatic
sarcoma. In older cysts the hydatid fluid is replaced by a jelly-like mass.
Clinical Disease
Cysts form primarily in the liver and growth in the vena cava or portal vein
results in metastases in the lung or brain. Clinical disease is similar to that of E.
granulosus.
Diagnosis
1. Laboratory diagnosis is can be made by ELISA.
2. Clinical diagnosis is made by ultrasound.
Table 6
Differences between the hydatid cysts of E. granulosus and E. multilocularis
The Cestodes
The cestodes (or tapeworms) form a group of worms, exhibiting two unmistakable mor-
phological features; they all possess flat, ribbon like bodies and lack an alimentary canal.
Adult tapeworms usually inhabit the alimentary canal of their hosts (though they
occasionally are found in the bile or pancreatic ducts) and attach themselves to the
mucosa by means of a scolex. Despite the lack of a digestive system they do absorb
food from the hosts intestine; thereby providing the tapeworms a habitat that is
associated with high nutritional levels, feeding the tapeworms high growth rate.
Larvae on the other hand show a wide range of habitat preferences, being found
in almost any organ of both vertebrate and invertebrate hosts. Though most larval
species show a preference for a particular site.
This lack of an alimentary canal markedly separates tapeworms from nematodes
and trematodes. The outer tegument of the body must serve not only as a protective
coating but also as a metabolically active layer through which nutritive material can be
absorbed, along with secretions and waste material to be transported out of the body.
79
 The body consists of a chain of segments or proglottids, which can be immature,
mature or gravid; the latter of which contain a fully developed uterus packed with
eggs. Therefore, each tapeworm is made up of a ‘string of individuals’ having a
complete set of reproductive organs in progressive degrees of sexual maturity and
budding off from a body attached to the host tissue by a head or scolex.
Except for Hymenolepis nana, which can develop directly in the same host, the
lifecycle of tapeworms involves both an intermediate and definitive host.
Taenia species
Introduction
Taenia species are the most common cestode parasites of humans. More than 60
million people are infected with T. saginata (“beef” tapeworm) world wide and
about 4 million are infected with T. solium (“pork” tapeworm). T. saginata has a
comsmopolitan distribution, but is more common in developing countries where
hygiene is poor and the inhabitants have a tendency of eating raw or insufficiently
cooked meat. T saginata is the most common adult tapeworm found in man. T solium
is virtually extinct in Europe and the USA.
The adults of both species live in the small intestine of man, the definitive host (Fig.
20). The gravid segments are very active and escape through the anus, releasing large
numbers of eggs in the perianal region or on the ground where they can survive for long
periods. When ingested by pigs or catle, the eggs hatch, each releasing an oncosphere,
which migrates through the intestinal wall and blood vessels to reach striated muscle within
which it encysts, forming cysticerci. When inadequately cooked meat containing the cysts
is eaten by man, the oncospheres excyst, settle in the small intestine and develop there into
adult cestodes over the next 3 months or so. The segments of T. solium are somewhat less
active than those of the beef tapeworm but its eggs, if released in the upper intestine, can
invade the host (auto-infection), setting up the potentially dangerous larval infection known
as cysticercosis in muscle of any other site. (Peters & Gilles, 1995)
Both humans and cattle or pigs are necessary to the complete life cycle of Taenia species.
(In Europe and the USA cattle are the normal intermediate hosts, but in the tropics several
other ruminants, e.g. goat, sheep llama and giraffe, may serve as the intermediate hosts.) Eggs
ingested by the intermediate hosts usually contain oncospheres. The oncospheres then hatch
out in the duodenum, pass into the intestine where they penetrate the intestinal wall and are
then carried by the circulation to be deposited in tissues (usually muscle). There they develop
into cysticerci larva, which are white and ovoid, measuring approximately 8 x 5mm.
Humans become infected by ingesting inadequately, cooked beef or pork with cysticerci,
containing an invaginated protoscolex. The protoscolexes evaginate and pass into the small
intestine where they attach themselves to the mucosa and develop into adult worms.
Eggs and proglottids are passed out in the faeces, and are then eaten by the
intermediate host, thus, perpetuating the life cycle.
Morphology
Ova of Taenia species are spherical, yellowish brown and measure 31-34mm in
80
Hymenolepis nana
Introduction
Hymenolepis nana, the dwarf tapeworm, is the smallest tapeworm to infect
humans. This cestode belongs to a large family known as Hymenolepididae. The
diagnostic features of this family are: scolex armed with one circlet of five hooks;
1-3 large testes and sacciform uterus. In addition to the H.nana, three other species,
H. diminuta, H. microstoma and H. citelli have been used extensively for studies on
cestodes.
Hymenolepis nana has a cosmopolitan distribution and is thought to be the most
common tapeworm throughout the world. The infection is more frequently seen in
children although adults are also infected, causing hymenolepiasis.
Life cycle
The lifecycle of H. nana does not require an intermediate host; complete
development occurring within the villi of a single host, resulting in a ‘direct’ life
cycle. Though it can also utilise an insect as an intermediate host.
The eggs that are released from mature proglottids in the upper ileum are usually
passed out in the faeces. If swallowed by another human they develop into hexacanth
oncospheres and burrow into the villi of the small intestine. This is where they develop
into tailless cysticercoids and then migrate towards the ileum and attach to commence
the formation of proglottids. The eggs, which are ingested by insects, such as fleas,
beetles or cockroaches hatch to form tailed cysticercoids, which remain unmodified
as long as they are inside the insect. If they are accidentally swallowed by a human
they pass down to the ileum and establish themselves. (Peters & Gilles, 1995)
Morphology
The egg containing the oncosphere bears three pairs of hooklets and is surrounded
by a membrane. This membrane has 2 polar thickenings from which arise threadlike
filaments extending into the space between the membrane and the colourless hyaline
shell, unlike those of H. diminuta, which do not possess any filaments.
The adult tapeworm is normally 2.5-4cm long. The scolex is knob like in shape,
has a rostellum with hooklets and 4 suckers. The segments are wider than they are
long. Ova are spherical or ovoid measuring 30-47mm in diameter. This is what
distinguishes it morphologically from H. diminuta.
Clinical Disease
Infections due to H. nana may cause no symptoms even with heavy worm burdens.
However, symptoms of restlessness, irritability, anorexia, abdominal pain and
diarrhoea have been reported. Heavy worm burdens may be caused by autoinfection,
which can be a problem in the immunocompromised.
Laboratory Diagnosis
Intersep has a comprehensive range of stains, fixatives and reagents for
parasitology use.
Preservation of parasites in faecal samples is not only important for maintaining para-
site structure during transportation but also as a means of preserving parasites for future
85
 Western Blots
Various Immunodiagnostic tests appear to give good results on serum or CSF.
Diagnosis using an immunodiagnostic test can be achieved using an in vitro
qualitative assay for the detection of IgG antibodies in serum reactive with T. solium
antigens present on a membrane.
Infected individuals develop a predominately IgG response to the parasite.
ELISA has been used as a screening test, but low sensitivity and frequent
artifactual crossreactions, or crossreactions with antibodies from other parasitic
infections, limit its usefulness as a confirmatory diagnostic test. The Western
Blot assay (U.S Patent No. 5,354,660) developed by Tsang et al, at the U.S.
Centers for Disease Control has been shown to provide a reliable method for
evaluation of sera from patients with clinically diagnosed active cysticercosis.
Field studies support a sensitivity of 98% and specificity of 100% for this
assay.
This assay is known as the QualiCode Cysticercosis Kit, the principle
behind the test is that it is a qualitative membrane-based immunoassay
manufactured from T. solium proteins. The T. solium proteins are fractionated
according to molecular weight by electrophoresis on a polyacrylamide slab
gel (PAGE) in the presence of sodium dodecyl sulfate (SDS). The separated
T. solium proteins are then transferred via electrophoretic blotting from the
gel to a nitrocellulose membrane. This antigen-bearing membrane has been
cut into strips for testing of individual samples. Sera are tested at 100X
dilution.
Intersep is now the leading suppliers of Qualicode Western Blot Kits.
Intersep now supply the kits for Cysticercosis, Hydatid disease, Babesiosis,
Human Granulocytic Ehrlichiosis, Canine and Human Lymes disease.
The Qualicode line of immunoassay kits is used to detect the presence of antibody
(either IgG or IgM) to a specific infectious disease agent. These qualitative assays
have the sensitivity and specificity of confirmatory tests. The easy to use kits are
Figure 20. Cysticercosis
designed for batch testing any number of samples from 1-24. Interpretation is
simplified by including Reference Strips, Positive and Negative Controls and a Record diameter. The shell is thick and radially striated. Within the shell, the onchosphere
sheet with every kit. has 3 pairs of hooklets. However, the microscopical appearance of the ova of
All reagents to perform the assay are provided along with three 8-channel T. saginata and T. solium are identical.
incubation trays. The length of the adult T. saginata is 4-8 meters long and that of T. solium is 3-
During the procedure, the strips containing the T. solium proteins are incubated 5 metres long. The proglottids of Taenia species can be identified by the number of
with serum specimens and washed to remove unbound antibodies. Visualisation of uterine branches; 7-13 for T. solium and 15-20 for T. saginata. If the scolex is recov-
human immunoglobulins specifically bound to T. solium proteins is performed by ered, the 4 suckers and rostellum of hooklets of T. solium will distinguish it from T.
sequential reaction with goat anti-human immunoglobulin-alkaline phosphatase saginata, which has 4 suckers but no hooklets.
conjugate and BCIP/NBT substrate. Bands corresponding to the positions of the Clinical Disease
resoled T. solium proteins will be visualised on the strip, indicating the presence in The presence of the adult worm rarely causes symptoms apart from slight ab-
the serum sample of IgG antibodies direct against Taenia antigens. Band positions dominal irritation with diarrhoea, constipation or indigestion. The accidental inges-
are compared to those on a reference strip developed using the cysticersosis positive tion of the embryonated ova of T. solium may result in cysticercosis in man (Fig. 21).
control. An infection due to an adult Taenia, in man or animals, is referred to as taeniasis.
84 81
 Figure 21. Taenia solium Figure 22. Taenia saginata
Table 7
T. saginata (the “beef” tapeworm) does not cause human cysticercosis. Comparison of T.saginata and T.solium
When the embryonated eggs are ingested, the embryos hatch out, migrate through
the intestinal wall and are carried around the body in the circulation and deposited in
various tissues (Fig. 22). Muscle and subcutaneous tissues are usually infected, but
cysticerci can infect most organs and tissues. Human cysticercosis is usually
asymptomatic unless the infection is particularly heavy or cysticerci are formed in
some vital area e.g. the brain, resulting in neurological sequelae.
Laboratory diagnosis
Diagnosis of intestinal taeniasis can be made by recovery of the characteristic
* No universal agreement to the number of uterine branches in these 2 species. As a rough
ova in the stool. However, the ova of T. solium and T. saginata are identical and guide, specimens with more than 16 branches are likely to be those of T. saginata and
diagnosis is made by the recovery of the segments or scolex. those with less than 10 branches are ikely to be of T. solium.
The diagnosis of cysticercosis depends upon serology. MRI scans may reveal the presence
of lesions in the brain. Calcified cysticerci are less often seen in the brain: in about one-third on surgical specimens. Calcification in muscles usually appears 3-5 years after initial infec-
of cases, 10 years or more after infection. Occasionally, the diagnosis is made histologically tion, and are most typically seen as spindle-shaped calcifications, most numerous in the thighs.
82 83