SPOROZOA - Coccidian protozoa

- Obligate intracellular pathogen that causes a mild diarrhea in

 Coccidia humans and animals
 Cryptosporidium sp. - Primarily infects small intestine of humans and mammals
 Isospora belli - Sporulated thick-walled oocysts are infectious and are
 Sarcocystis: resistant to many chemical disinfectants.
 S. hominis - Forms oocysts
 S. suihominis - Resistant to disinfection
 S. lindemani - Killed by ozone used in water treatment process,
 Toxoplasma gondii - Desiccation (sprayed) or by 5% ammonia solution
 Microsporidia
 Pneumocystis
 Microsporidia
 Pneumocystis carinii

Cryptosporidiasis
Classification

 Phylum – Apicomplexa (Sporozoa)

Life Cycle
 Class – Sporozoasida
 Subclass – Coccidiasina
 Order – Eucoccidiida
 Suborder – Eimeriina
 Family - Cryptosporidiidae.

- Cryptosporidium spp. has monoxenous life cycles, where all stages of

development (asexual and sexual) occur within one host.

1.) Cryptosporidium parvum; C. hominis

Crypto begins its life cycle as sporulated oocysts:  Hospitals (nosocomial) in HIV wards and pediatric
hospitals
(1) Which enter the environment through the feces of the infected
host. There is some evidence that it can also be spread by respiratory Epidemiology
secretions. – Cryptosporidium can be found worldwide. Cryptosporidiosis
(2) The infective oocysts reside in food and water. Most transmission is widespread geographically.
occurs through recreational water use, such as in pools and – Cases typically increase during the summer with increased
lakes. Crypto has also entered drinking water supplies, causing major use of recreational water.
outbreaks in cities around the world. Oocysts are resistant to chlorine
Morbidity or Mortality
treatment, presenting a major water purification problem during
 In most country
outbreaks. They can also withstand freezing temperatures. - 2% of population infected
(3) Infection occurs when the oocysts are ingested by a suitable - 80% previously exposed
host. While in the intestines, the oocyst releases sporozoites which  Worldwide
invade the epithelial linings of the intestines or the lungs (depending – Prevalence is (1 to 4.5%) lower in developed countries,
on the method of transmission). Within these cells, the sporozoites – Higher in developing countries (3 to 20%)
- The infection is usually self-limiting in healthy people and they
undergo schizogony, or asexual reproduction. The sporozoites then
usually recover after 2 to 4 days. Episodes of diarrhea can last
enter a sexual reproductive stage. Female macrogamonts and male 1 to 4 weeks
microgamonts develop, and fertilization occurs. The resulting zygote  Animals
can develop into: a) a thick-walled oocyst that will exit the host, or b) - Morbidity high, mortality low
a thin-walled oocyst which will autoinfect the host. - Animals that are immunosuppressed and neonatal animals are
The infectious oocysts are passed through the feces (1). These oocysts more susceptible to severe disease.
enter the environment to await ingestion by the next host.
Transmission
• Fecal-oral route
History  Person-to-person
 Person-to-animal
 1912
 Waterborne
- Discovered by Earnest Edward Tyzzer (American
 Foodborne
parasitologist)
 Mechanical
- Discovery of C. parvum is now recognized as the parasite
- Oocysts are shed in the feces and are immediately infectious. If
usually responsible for mammalian cryptosporidiosis
in a moist environment, the shed oocysts can survive for 2 to 6
- In 1970’s, it was not clear whether C. parvum was an
months. Direct transmission between animals and humans is
opportunistic parasite or a pathogen.
common. It is estimated that 50% of dairy calves shed oocysts;
- Outbreaks associated with:
calves can then spread cryptosporidiosis to each other or to
 Drinking water, food
humans. Transmission of cryptosporidiosis from household
 Swimming pools and lakes
pets to humans can also occur but is rare and poorly
 Unpasteurized apple cider
 documented. Additionally, humans and animals can ingest the
organism through eating or drinking contaminated water or
food
• Aerosol (rare)

ANIMALS AND CRYPTOSPORIDIOSIS

Disease in Animals
- Cryptosporidium parvum Not host specific and can infect all Diagnosis
mammals  Differential diagnosis should include:
- Common in young calves and lambs and other ruminants – Giardia, Isospora
- Infections occur in neonates – Microsporidia
- Other species: – Salmonella
- Pigs – Rotavirus
- Dogs (rare) – Other diarrheal diseases
- Cats (rare)  Diagnostic tests
- Horses (rare) – Cryptosporidiosis can be diagnosed by finding C.
- Other species of cryptosporidium can affect birds and reptiles parvum after a Fecal flotation using sucrose or zinc
- Incubation period: ~4 days in calves sulfate solutions
- Clinical signs of cryptosporidiosis: – The mature oocysts are 4-5 µm in diameter and contain
 Anorexia four thin, flat, motile sporozoites. Cryptosporidium
 Diarrhea parvum can also be detected through an Acid-fast
 Weight loss staining
 Tenesmus – Oocysts are not shed continuously and repeated
- More severe disease can occur with concurrent infections. sampling may be necessary. The oocysts appear red
Animals can also be colonized without symptoms. after the staining.
 Immunofluorescence - used to detect in feces
Post Mortem Lesions  Cryptosporidiosis can be detected by stained
 Gross lesions biopsy/necropsy specimens or fresh intestinal scrapings.
- Hyperemia of intestinal mucosa
- Yellowish intestinal contents
- Affect most of the intestinal tract
- Most severe lesions are found in the distal small intestine
 Microscopic lesions
- Mild to severe villous atrophy
- Spherical organisms in the brush border
Treatment - Immunofluorescence and by staining a biopsy specimen
- No treatment (supportive care) (occasionally done this way).
- Disease tends to be self-limiting. No vaccine has been - No treatment
developed - No vaccine

HUMANS AND CRYPTOSPORIDIOSIS Prevention and Control

 Prevention in Animals
Disease in Humans - Keep sick away from healthy animals is a good way to limit
– Similar to disease in animals exposure
– Parasite is accidentally swallowed - common in the environment and can be carried by
– Contaminated water, food, hands, stool or unwashed animals without any symptoms
hands. - Clean and disinfect
– People affected: - Chlorine does not effectively kill the organism and
 Children or young humans it is resistant to many disinfectants. Clean and
 Immunosuppressed (AIDS patient are more disinfectant (5% ammonia solution can work) areas
susceptible) where sick animals have been or had diarrhea
 Travelers - Good nutrition to the animals and keep the animals
 Swimmers healthy to minimize the risks.
- Incubation period: 1 to 12 days with 7 days being typical
- Clinical signs is similar to animals  Prevention in Humans
 Profuse, watery diarrhea with cramping - Wash hands
 Abdominal pains - Wash fruits/vegetables
 Anorexia - Don’t swallow water from:
 Flatulence and Malaise - Lakes
- Some individuals can also experience vomiting, weight loss, - Streams
fever, or myalgia. - Public pools
- Self-limiting in healthy people - Do not swim in public pools if you have recently had
- Chronic, debilitating and severe and may become serious in diarrhea.
immunocompromised people - Minimize contact with young animals
- Asymptomatic infections can also occur.

Diagnosis and Treatment 2.) Isospora belli

 Differential diagnosis
- Similar to animals and include diseases that cause Isosporiasis
diarrhea - a.k.a cystoisosporiasis is a human intestinal disease caused by
 Diagnostic tests the parasite or agent Isospora belli (now known
- Fecal flotation with sucrose or zinc sulfate as Cystoisospora belli)
- Acid-fast staining (oocysts appear red) - Agent – Isospora belli
 - Worldwide – tropical and subtropical - Swallowing of mature parasite
- Infection occurs in Immunocompromised individuals - Contaminated food and water
notably AIDS patients, and outbreaks have been reported in - The infected host then produces an immature form of the
institutionalized groups in some countries. parasite in their feces, and when the parasite matures, it is
- First documented in 1915 capable of infecting its next host
- Usually spread indirectly – thru contaminated food or water - Poor hygiene and sanitation

Classification
• Domain: Eukaryote
• Phylum: Apicomplexa (Sporozoa)
• Class: Coccidia
• Order: Eucoccidiorida
• Family: Eimeriidae
• Genus: Isospora
• Species: I. belli

Signs and Symptoms

– Acute, non-bloody diarrhea
– Crampy abdominal pain lasts for weeks and result in
Malabsorption
– In immunodepressed patients, and in infants and children, the
diarrhea can be severe.
– Anorexia and weight loss
– Eosinophilia may be present (differently from other protozoan
infections)
– Vomiting - At time of excretion, the immature oocyst contains usually
– Fever one sporoblast (more rarely two). In further maturation after
excretion, the sporoblast divides in two, so the oocyst now
Sites of Infection contains two sporoblasts. The sporoblasts secrete a cyst wall,
- Intestinal villus (Enterocytes) thus becoming sporocysts; and the sporocysts divide twice to
- Lamina propria of the villus produce four sporozoites each. Infection occurs by ingestion of
- The coccidian parasite Isospora belli infects the epithelial cells sporocyst-containing oocysts: the sporocysts excyst in the small
of the small intestine (intestinal villus and lamina propria), intestine and release their sporozoites, which invade the
and is the least common of the three intestinal coccidia that epithelial cells and initiate schizogony. Upon rupture of
infect humans (Toxoplasma, Cryptosporidium, and Isospora). the schizonts, the merozoites are released, invade
new epithelial cells, and continue the cycle of asexual
Transmission multiplication. Trophozoites develop into schizonts which
- Oral-fecal route contain multiple merozoites. After a minimum of one week, the
 sexual stage begins with the development of male and female • Kingdom: Chromalveolata
gametocytes. Fertilization results in the development of • Phylum: Apicomplexa
oocysts that are excreted in the stool. Isospora belli infects both • Class: Canoidasida
humans and animals. • Order: Eucoccidiorida
• Family: Sarcocystidae
Diagnosis • Genus: Sarcocystis
 Typical Lab Analyses:
- Microscopy Causal Agents
- Morphologic comparison with other intestinal parasites  Sarcocystis hominis
- Bench aids for Isospora  Sarcocystis suihominis
 Microscopic demonstration of the large typically shaped - (both S. Hominis and S. suihominis) use humans as
oocysts is the basis for diagnosis. Because the oocysts may be definitive hosts and are responsible for intestinal
passed in small amounts and intermittently, repeated stool sarcocystosis in the human host. Humans may also become
examinations and concentration procedures are dead-end hosts for non-human Sarcocystis spp. after the
recommended. If stool examinations are negative, examination accidental ingestion of oocysts.
of duodenal specimens by biopsy or string test (Enterotest) may  Sarcocystis lindemanni
be needed. The oocysts can be visualized on wet mounts by - probably humans are only accidental hosts, since many
microscopy with bright-field, differential interference contrast species of monkeys show such cysts, which may induce
(DIC), and epifluorescence. They can also be stained by myositis. This parasite apparently, has a worldwide
modified acid-fast stain. distribution, since it was originally described by the
German physician Lindemann, who, in the service of the
Prevention Russian Czar, found these cysts in the tongue‐muscles of
 Avoid contaminated food and water hanged prisoners.
 Good hand-washing
 Personal-hygiene practices Life Cycle

Treatment
- Prescription antibiotics
- Drug of choice: trimethoprim-sulfamethoxazole

3.) Sarcocystis

Sarcocystosis - caused by species of Sarcocystis, an intracellular protozoan

parasite in the phylum Apicomplexa. . . . . .
- The name Sarcocystis is dervived from Greek: sarx = flesh
and kystis = bladder.

Classification

- Both sporulated oocysts (containing two sporocysts) and
individual sporocysts can be passed in stool (1). Sporocysts
contain four sporozoites and a refractile residual body.
Sporocysts ingested by the intermediate host (cattle for S.
hominis and pigs for S. suihominis) rupture, releasing
sporozoites. Sporozoites enter endothelial cells of blood vessels
and undergo schizogony, resulting in first-generation schizonts.
Merozoites derived from the first-generation invade small
capillaries and blood vessels, becoming second-generation
schizonts. The second generation merozoites invade muscle
cells and develop into sarcocysts containing bradyzoites, which
are the infective stage for the definitive host(2). Humans
become infected when they eat undercooked meat containing
these sarcocysts (3). Bradyzoites are released from ruptured
cysts in the small intestine and invade the lamina propria of the
intestinal epithelium(4). There, they differentiate into macro-
and microgametocytes. Fusion of male and female gametes (5)
results in the formation of oocysts (6). Oocysts sporulate in the
intestinal epithelium and are shed from the host in feces(7).
Due to the fragile nature of the oocyst wall, individual
sporocysts may also be detected in feces.
Geographic Distribution
- Worldwide
- Human intestinal sarcocystosis has been reported worldwide.
Symptomatic cases of human muscle sarcocystosis have been seen
mainly in Southeast Asia, probably due to the distribution of a
definitive host. Sarcocystis spp. have also been found in human
muscle tissues in the U.S., often as an incidental finding at autopsy.
- Common in areas where livestock is raised.
Incubation Period
- In humans, the clinical signs occurred as soon as 3 to 6 hours in
the intestinal form, then recurred 14 to 18 days later.
- Incubation period for the muscle form is unknown.
Clinical Signs
 Humans as an Intermediate Host (IH)
- Myositis (primary syndrome)
- Spectrum of illness ranges from acute self-limited
infections to chronic, moderately severe disease
- Painful muscle swelling and erythema
- Muscle tenderness, weakness and fever
- Respiratory problems:
- Bronchospasm and cough
- Vasculitis:
- Arthralgia and pruritic rashes
- Headache and malaise
- Lymphadenopathy and muscle wasting
- Chronic cases can have persistent or recurrent symptoms for up
to seven years. Many infections may be asymptomatic. In these
cases, humans are dead-end intermediate hosts.
 Humans as a Definitve Host (DH)
- (S. suihominis or S. hominis),
- Intestinal sarcocystosis:
- Fever, chills and sweating
- Diffuse abdominal tenderness, diarrhea
- Dehydration occur as a result of Nausea and vomiting
- Eosinophilic enteritis
- Rare cases of acute intestinal obstruction
- Intestinal sarcocystosis is transient and usually self-limited;
chronic enteritis has not been described. Many or most cases
are thought to be asymptomatic.
Communicability
- Sarcocytis hominis – cattle by sarcocysts shed in the feces
- Sarcocystis suihominis – pigs by sarcocysts shed in the feces
- Sarcocystis lindemanni – monkeys show cyts
- Shedding begins after 10 to 13 days and can continue for up to
six months. Humans serving as the intermediate host cannot
spread the infection to others.
Diagnostic Tests
• Zinc sulfate floatation - Intestinal infections can be diagnosed
by detecting sporulated sporocysts in the feces
• Microscopy - Sarcocysts may be found in the muscles by
microscopic examination of a muscle biopsy
• CT scan or MRI - visualize sarcocysts in the muscles
• CBC – reveal eosinophilia
• Indirect Immunofluorescent antibody test
• Immunoblotting - useful for muscle infections, but is not considered
to be valuable for intestinal infections. Serologic tests may not be
widely available.
- More than 60 million people in the United States probably carry
the Toxoplasma parasite, but very few have symptoms because
the immune system usually keeps the parasite from causing
illness. However, expectant mothers should be cautious
because an infection can cause problems in pregnancy.
- Image pictured is of brain tissue with abnormal growth due to
toxoplasmosis

- Can be transmitted by ingestion of either oocysts (in the feces

of cats) or bradyzoites (in raw or undercooked meat).

Classification
Treatment • Domain: Eukaryote
- Intestinal sarcocystosis – may not be treated as the typical • Phylum: Apicomplexa
infection is self-limiting • Class: Conoidasida
- Symptomatic myositis – antiparasitic agent (metronidazole, • Order: Eucoccidiorida
Cotrimoxazole and albendazole) • Family: Sarcocystidae
- Corticosteroid to reduce inflammation • Genus: Toxoplasma
- Asymptomatic cases may not require treatment. • Species: T. gondii

Prevention Causation
- Avoid eating raw or undercooked beef - Coccidian, obligate, intracellular parasite
- Freezing the meat to -20 degrees C. - Responsible for zoonotic infections in man and other mammals
- Avoid food contaminated feces - Generally causes very benign disease in immunocompetent
- Good personal hygiene adults
- The definitive host is not known in many cases, particularly - T. gondii is one of the most common human parasite in the
those that occur in Southeast Asia. world. The organism was first discovered in the gundi, a North
African rodent.
- Tissue cyst forming coccidia
4.) Toxoplasma gondii - Felines are definitive host
Toxoplasmosis - infects wide range of birds and mammals (intermediate
- It is an infection caused by the parasite Toxoplasma gondii hosts)
-
 Definitive Host  Oocysts
• adult forms - Only members of the cat family shed oocysts
• sexual reproduction - Cats become infected by ingesting either oocysts from fecal
Intermediate Host contamination or tissue cysts present in flesh of eaten
• immature forms animals. Digestive enzymes release the organisms, which
• asexual reproduction invade the feline small intestine. The organisms reproduce,
producing millions of noninfectious, unsporulated oocysts
that are excreted in the feces of cats for up to 2 weeks.
Life Cycle Once outside of the cats body, sporogony occurs for up to
• Two phases: 21 days, resulting in the development of infectious oocysts.
1. Intestinal - Occurs only in cats (wild and domesticated In, warm moist soil these oocysts can remain viable for
cats), and results in the production of oocysts. more than 1 year.
2. Extra - intestinal - occurs in all infected animals
(simultaneously with the intestinal phase in cats), and
results in the production of tachyzoites and, eventually,
bradyzoites.

 Trophozoites
- Tachyzoites are the actively proliferating trophozoites,
observed during the acute stage of infection
- Thachyzoites can be found in any organ but occur most
commonly in the brain, skeletal muscle, and heart muscle.
Intracellular infection can occur in all mammalian cells
except anuclear erythrocytes. Intracellular multiplication
continues until host cells lyse or a tissue cyst is formed. In
an immunocompetent host, tachyzoites are eliminated
and tissue cysts form

 Tissue Cysts
- found most commonly in the brain and in skeletal and cardiac
muscle but can occur in any organ
- These cysts contain slowly growing trophozoites known as
bradyzoites. In an immunocompromised host, tachyzoite
replication results in development of focal necrosis, such as
necrotizing encephalitis, pneumonitis, or myocarditis. Tissue
cysts usually are observed during the chonic or latent stage of
infection, causing little, if any, inflammatory response. These
cysts can be involved in disease transmission if present in
tissues ingested by carnivores. After ingestion, cysts are broken
down by digestive enzymes which release the organsims,
allowing them to invade the GI tract where they spread via
blood and lymphatics.
Signs and Symptoms
- Most people who are infected are not aware of it
- Fever
- Sore throat
- Sore muscles
- Tiredness
- Swollen glands in neck, armpits and groin
- Temporary blurred vision or loss of vision
- Most infants infected in the womb show no signs at birth but
develop symptom later on in life. A small percentage of
infected newborns have serious eye or brain damage at birth.
Diagnosis
- If a health care provider suspects that a pregnant woman has
an active Toxoplasmosis infection he or she may recommend
one or more of several available blood tests. Blood tests are
used to detect for microscopic parasites and to measure the
body’s level of antibodies
- Serological tests
- Polymerase chain reaction (PCR)
- Histological demonstration of the parasite and or its antigens
(i.e. immunoperoxidase stain)
- Isolation of the organism
Treatment
• Treatment should be developed in consultation with an
obstetrician and infectious disease specialist
• Children born with congenital toxoplasmosis are treated with
different combinations of anti-toxoplasmosis medications (plus
leurovorin) usually one year after birth
• In healthy older children who develop severe Toxoplasmosis
infections, treatment usually last for 4 to 6 weeks (or at least 2
weeks after symptoms are gone).
• Hospitalization may be necessary for those weak immune
systems
• Those with AIDS may need to take anti-toxoplasmosis
medication for life.
Prevention
• Wear gloves when handling soil
• Wash hands with soap and water after outdoor activities
• When preparing raw meat, wash any cutting boards, sinks,
knives and other utensils that touched the raw meat thoroughly
with soap and hot water to avoid contaminating other foods
• Cook all meats thoroughly
• Keep cats indoors and feed them dry or canned food rather
than allowing them to have access to wild birds and rodents
• Clean litter box everyday (because the parasite found in cat
feces needs one or more days after it is past to become
infectious)
• Thoroughly wash hands after cleaning litter box
5.) Microsporidia
• Eukaryotic, unicellular organism belonging to the phylum
Microspora
• All are Obligate, spore-forming, intracellular parasites that
invade vertebrates and invertebrates.
• Feature: Polar tube or polar filament found in the spore used to
infiltrate host cells
• They are widely distributed in nature with over 1200 species
characterized. However, microsporidia have only recently been
documented to parasite humans, and more research is needed
to understand this emerging infectious disease.
• Found in patient with compromised immune system especially
those infected with Human Immunodeficiency Virus (HIV) or
have undergone organ transplants.
• Parasites in fisheries, veterinary medicine and pest
management
Classification and Taxonomy
• Initially thought to be protozoan (kingdom Protista)
• Recent DNA studies classified Microspora as fungi kingdom or
at least as a sister kingdom to Fungi.
• Class, order and family within the Microspora phylum are
frequently revised and debated
• Traditionally, species were identified by observing the physical
characteristics of the spore, life cycle and relationship with the
host cell. However, recent scientific studies using genetic tools
(namely ribosomal RNA sequencing) have challenged this
approach and suggest genetic markers a more correct method
for scientific classification. There are now over 1200 species
identified in 143 genera.
• Currently, 14 species in 8 genera known to infect humans
Phylum Microspora
History of Discovery
• Discovered in the late 1800’s
• First human case in 1958 in a Japanese child
• Microsporidiosis associated with the arrival and spread of and
HIV
• microsporidiasis is primarily found in patients with AIDS or are
otherwise immuno-compromised (like organ transplant
patients)
• Nosema and Brachiola documented in immune-competent
patients
• Casual, accidental or opportunictic agents in humans
Agent Morphology
- Microsporidia are primitive eukaryotes with well defined nuclei
and plasma membrane but lack some typical organelles found in
 more typical eukaryotes mainly mitochondria, stacked golgi and
peroxisumes.
- Microsporidia spores are all round and oblong and those
associated with human infection tend to be about 1-4 µm in size.
All have a characteristic coiled polar tube, tubule or filament,
layered polarplast, a posterior vacuole (thought to function as
golgi) and a protective exospore made of proteins and chitin.
Chitin is responsible for the spores’ high environmental
resistance.
- During infection, the cell membranes of the spore (also called the
sporoplasm) is injected into a host cell though osmosis
pressure. After the infection, the microsporidium relies on the
host cell for energy and begins to multiply inside the host cell
cytoplasm. Species differ in their relationships with host cell;
some species alter host cell functioning to induce more nutrient
absorption and cell growth to accommodate the agent.
- Microsporidia may reproduce sexually or asexually. In asexual
reproduction, nuclear division takes place and form one or more
pairs of nuclei, and cellular division may isolate the nuclei or pair
them in a diplokaryon arrangement. Sexual reproduction is not
well understood but it thought to involve autogamous fusion and
reorganization of genetic material
Life Cycle
(1) the environmentally resistant, infectious spore is ingested or
otherwise contracted. This environmental stimulus activates
germination in the spore, releasing the polar tubule through eversion.
(2) The spore then inject the polar tubule into a host cell and
(3) release their sporoplasms into host cells.
(4) The microsporidia meronts then multiply either in contact with the
host cytoplasm.
(5) They then undergo sporogony to further divide and form
sporoblasts, preparing the cell for thick exospore layers characteristic
of mature spores. The spores can either be dispersed freely within the
host cell cytoplasm or packaged in sporophorous vesicles.
Finally, (6) when spores completely fill the host cell, the plasma
membrane is affected and releases the spores to its surroundings. The
spores may then infect other surrounding cells, be transported to new
sites within the host, or be excreted in feces or urine to infect other
hosts.
The exact incubation period for microsporam spores is unknown, but
the spores are considered extremely resistant and thus assumed to
persist in the environment for long periods of time.
Reservoirs and Vectors
• Ubiquitous and are capable of infecting any animal cells
including those of insects, fish, mammals and even other
parasites
• Common in anthropods
• 14 species infecting humans are found in a number of wild and
domesticated mammals (e.g. rabbits, mice, puppies, kittens,
etc
• No specific vectors for microsporidiosis in the standard sense
(infected insects may infect humans if they are ingested
Transmission
• Inhaling
• Ingesting
• Contracting spores
• Ocular or sexually transmitted
• Transmitted in water
 • Significant contact with infected animals may also transmit the
disease (zoonoic infection) but cases are ingested
Clinical Presentation
- Chronic diarrhea and wasting are the most common
symptoms of microsporidiosis, but different species invade
different sites including the cornea, binary track and muscles.
Thus, the symptoms of microsporidiosis varies greatly
depending on the site of infection.
Diagnostic Tests
• ID of spores in feces, urine, other body fluid or body tissues
• Transmission electron Mx (gold standard) for identifying
specific species and diagnosing microsporidiosis but is often too
expensive and time consuming
• Microscopy using various stains including gram stains
(microsporidia spores are gram-positive and stain dark violent
and become readily visible under the microscope), modified
trichrome stains (e.g. trichrome blue), Warthin-Starry silver
stains, Giemsa, and chemofluorescent agents like Calcoflur.
• Immunofluorescent assays
• Molecular techniques
Treatment and Management
• Albendazole - most common for all species
• Topical Fumagillin for ocular infections. However, most drug
treatments will not fully eradicate the parasites
• Nikkomycin Z (NIK-Z), a drug that inhibits chitin synthesis, has
been shown to be affective against a host of fungal pathogens.
• Metrinidazole
• Thalidomide
• Electrolyte replacement and fluids regularly and maintain
nutritional intakes for severe chronic diarrhea
Public Health and Prevention Strategies
- Water filtration - best preventative strategy. Measurement and
filtration techniques for microsporidia spores remain rudimentary
and underdeveloped, though the scientific community is actively
trying to amend this knowledge gap.
- No vaccines available
- Work-up of Microsporidiosis in immunocompromised patients
6.) Pneumocystis pneumonia
• Form of pneumonia caused by the yeast-
like fungus Pneumocystis jirovecii.
• previously classified as Pneumocystis carinii
• This organism is common in the environment and does not
cause illness in healthy people.
• In 1980’s, biochemical analysis of the nucleic acid
composition of pneumocysitis rRNA and mitochondrial
DNA identified the organism as a Unicellular fungus
Change of Nomenclature
• Pneumocystic carinii – Rats
• Pneumocystis jeroveci – Humans
Morphology
• Trophic form- trophozoite
• Cyst form- spore
• Spherical, Elliptical: 4-6 microns, contains 4-8 nuclei
• Extracellular pathogen
• The trophic forms can conjugate and form cysts that mature to
contain eight nuclei. These cysts rupture, freeing eight trophic forms
from the intracystic nuclei. Also, the trophic forms can replicate
asexually via binary fission
• Stained with Silver stain, toluidine blue, calcofluor white

Individual predisposed to P. jeroveci infection

- Can cause lung infection in people with a weakened
immune system to any of the ff conditions:
 Cancer
 Chronic use of corticosteroids or other medication that
affect immune system
 HIV/AIDS
 Solid organ and or bone marrow transplant
Life Cycle
- These fungi are found in the lungs of mammals where they Clinical Presentation
reside without causing overt infection until the host’s • Progressive exertional dyspnea (95%)
immune system becomes debilitated. • Fever (>80%)
- Then, an oftentimes lethal pneumonia can result. • Nonproductive cough (95%)
- Asexual phase: trophic forms replicate by mitosis to . • Chest discomfort
Sexual phase: haploid trophic forms conjugate and produce • Weight loss
a zygote or sporocyte (early cyst) . • Chills
- The zygote undergoes meiosis and subsequent mitosis to • Haemoptysis (rare)
produce eight haploid nuclei (late phase cyst) . Spores
exhibit different shapes (such as, spherical and elongated Differentiation of bacterial pneumonia & pneumocystis pneumonia
forms).
- It is postulated that elongation of the spores precedes
release from the spore case. It is believed that the release
occurs through a rent in the cell wall. After release, the
empty spore case usually collapses, but retains some
residual cytoplasm .
- A trophic stage, where the organisms probably multiply by
binary fission is also recognized to exist.
Diagnosis mediated immunity, and it is a major infection in patients with
• Examination of bronchoalveolar lavage the acquired immune deficiency syndrome (AIDS). Severe
• Lung biopsy protein-calorie malnutrition alone may provoke the disease.
• Examination of induced sputum Immunosuppressive drugs used for cancer or organ
• bronchoalveolar lavage, or needle aspiration of the lung. transplantation render the individual susceptible to P carinii
• Direct fluorescent method pneumonitis.
• PCR method
Treatment and Prophylaxis
• Trimethoprim-Sulfamethoxazole (TMP – SMZ) –
• Pintimidum isothionate
• Atovaquone
• Trimetrevate
 (TMP – SMZ) – preferred because of low toxicity and greater
efficacy
Stains used to examine
• Gomori
• Giemsa stain TACBSMTTWOA
• Fluorescence-labelled antibody
• Toludine blue O
• Methanamine silver
• Calcoflour white

Immunity
• In immunocompetent individual, P. jeroveci do not cause
infection
• Cell mediated helps in the protection from pneumocystis
Infections are prevalent until CD4 count drops below 400/
micro liters
• With rare exceptions, P carinni causes disease only when
natural mechanisms of host defense are compromised.
Pneumonitis tends to occur in patients with impaired cell-