Bio 310

Lecture 17
22nd February 2023
Eukaryotic Microbes
and
Parasites
Eukaryotic Microbes
 Protozoa
• Life Stages –
– Trophozoite -vegetative; feeding, mostly motile
– Cyst – dormant; protective thick wall
• Most are free living in water and soil
• Classified by motility & life cycle
• Subdivided by location in human host (GI, blood, GU)

1. Sarcodina- Amoeba - move by pseudopods

2. Ciliophora - Ciliates - move by cilia
3. Mastigophora - Flagellates - move by flagella
4. Apicomplexan - Sporozoa – complex life cycle
Diversity among Protozoa
Flagellates: INTESTINAL PROTOZOA
• Giardia lamblia
• Dientamoeba fragilis
• Chilomastix mesnili
• Trichomonas hominis unicellular eukaryotic organisms
• Enteromonas hominis
• Retortamonas intestinalis
Ameba:
• Entamoeba histolytica
• Entamoeba dispar
• Entamoeba coli
• Entamoeba hartmanni
• Endolimax nana
• Iodamoeba bütschlii
Apicomplexa:
• Cryptosporidium hominis
• Cryptosporidium parvum
• Cyclospora cayetanensis
• Isospora belli
Other:
• Blastocystis hominis
• Balantidium coli
Fecal-Oral Transmission Factors
• poor personal hygiene
• food handlers
• institutions
• children in day care centers
• developing countries
• highly endemic
• poor sanitation
• travelers diarrhea
• water-borne epidemics
• zoonosis
• Entamoeba = no
• Cryptosporidium = yes
• Giardia = controversial
Control/Prevention
• improve personal hygiene
• especially institutions
• treat asymptomatic carriers
• eg, family members
• health education
• hand-washing
• sanitation
• food handling
• protect water supply
• treat water if questionable
• boiling
• iodine
 Flagellate
• Trichomonas vaginalis
– no cyst stage
– Trichomoniasis - STI
• Giardia lamblia
– intestinal malabsorption
– Traveler's diarrhea, day
care centers, hikers

Figure 12.17b-d
Giardiasis

Giardia Enteritis
Lambliasis
Beaver Fever
 Overview
• Organism
• History
• Epidemiology
• Transmission
• Disease in Humans
• Disease in Animals
• Prevention and Control

Center for Food Security and

Public Health, Iowa State
 Organism
• Giardia intestinalis
– Protozoan parasite
• Also known as:
– Giardia lamblia
– Lamblia intestinalis
– Giardia duodenalis
• Isolated from humans, domestic animals,
and wild animals
Center for Food Security and
Public Health, Iowa State
 Organism
• Human infections
– Humans are main reservoir
• Interspecies/zoonotic transmission
– Importance of animal reservoirs unclear
• Non-zoonotic Giardia spp. found in:
– Rodents
– Birds
– Reptiles
– Amphibians
Center for Food Security and
Public Health, Iowa State
Giardia lamblia
• worldwide distribution
• higher prevalence in
developing countries (20%) CYST
• infective stage
• 1-6% in temperate countries • passed in feces
• most common protozoa
found in stools
• ~200 million clinical
cases/year
• giardiasis
• often asymptomatic
• acute or chronic diarrhea TROPHOZOITE
• fecal-oral life cycle • replicative stage
• small intestine
 Pathogenesis
• epithelial damage
• villus blunting
• crypt cell hypertrophy
• cellular infiltration
• malabsorbtion
• enzyme deficiencies
• lactase (lactose
intolerance)

Possible Mechanisms
• mechanical irritation
• obstruction of absorption
Adhesive Disk and Attachment
 History
• 1681
– van Leeuwenhoek, the “Father of
Microbiology,” observed Giardia trophozoites
in his own stool
• Doubt common regarding pathogenicity of
Giardia organisms
• 1970s
– Symptomatic travelers from Soviet Union
increased awareness
Center for Food Security and
Public Health, Iowa State
 Geographic Distribution
• Giardia intestinalis
– Occurs worldwide
– Most common in warm climates

Center for Food Security and

Public Health, Iowa State
Morbidity and Mortality: Humans
• Populations affected
– Children
– Travelers, hikers
– Swimmers
• Prevalence
in developed countries
– 2% of adults
– 6-8% of children
• Up to 15% in developing countries
Center for Food Security and
Public Health, Iowa State
Morbidity and Mortality: Humans
• Naïve populations
– Morbidity rate up to 20%
• Infections often resolve spontaneously
• Chronic infections occur
– May contribute to decreased lifespan in
immunodeficient individuals

Center for Food Security and

Public Health, Iowa State
 Parasite Stages
• Two stages of the parasite:cyst and
trophozoite

Center for Food Security and

Public Health, Iowa State
 Transmission
• Cysts
– Direct transmission
– Fomites
• Contaminated water and/or food
• Ingested cysts release trophozoites
• Trophozoites multiply and encyst in
intestines
• Excreted in feces

Center for Food Security and

Public Health, Iowa State
 Survival
• Cysts
– Survive well in cool, moist conditions
– Remain viable for months in cold water
• Two months at 8oC
• One month at 21oC
– Can also survive freezing
– Susceptible to desiccation and direct sunlight

Center for Food Security and

Public Health, Iowa State
Life Cycle
• Cysts responsible for
transmission
• Cysts and
trophozoites found in
feces
• Ingested by host
• Importance of animal
reservoirs unclear

Center for Food Security and

Public Health, Iowa State
 Disease in Humans
• Incubation period: 1-25 days
• Most infections asymptomatic
• Symptoms of clinical disease
– Mild to severe gastrointestinal signs
• Sudden onset diarrhea
• Foul-smelling stools
• Abdominal cramps
• Bloating, flatulence
• Nausea, fatigue
• Weight loss
Center for Food Security and
Public Health, Iowa State
 Disease in Humans
• Illness usually lasts for 1-2 weeks
• Chronic infections reported
– May last months to years
– Immuno-deficient and immuno-compromised
individuals
– May lead to malabsorption syndromes,
vitamin deficiencies, severe weight loss,
and debilitation
• Disaccharide intolerance
Center for Food Security and
Public Health, Iowa State
 Diagnosis
• Direct observation in feces
– Trophozoites
• “Tear drop” shape
• Two nuclei and tumbling mobility
– Cysts
• Approximately 13 microns long
• Oval, with 2-4 nuclei
• Immunofluorescence
• ELISA, PCR
Center for Food Security and
Public Health, Iowa State
 Treatment
• Anti-protozoal drugs
– Metronidazole
– Tinidazole
– Ornidazole
• Chronic cases
– May be resistant
– Prolonged therapy may be necessary

Center for Food Security and

Public Health, Iowa State
 Prevention and Control
• Water
– Do not drink contaminated water
• Untreated lakes, rivers, shallow wells
– Treat potentially contaminated water
• Heat (rolling boil for one minutes)
• Filter (absolute pore size of one micron)
• Chlorinate
• Food
– Wash raw fruits and vegetables
Center for Food Security and
Public Health, Iowa State
 Prevention and Control
• Practice good hygiene
– Hand washing
– Don’t swim in recreational
waters for at least two
weeks after symptoms end
– Avoid fecal exposure

Center for Food Security and

Public Health, Iowa State
 Prevention and Control
• Limit environmental contamination
– Clean and promptly remove feces
from surfaces
• Keep pets indoors
• Vaccination
– Dogs and cats
– Use is controversial

Center for Food Security and

Public Health, Iowa State
Diversity among Protozoa
 Amoeba
• Entamoeba histolytica
– Amoebic dysentery

• Naegleria
– primary amoebic
meningoencephalitis

• Acanthamoeba
– contact lens contaminant

Figure 12.18a
 Amoebae

• Protozoa with no truly defined shape

• Move and acquire food through the use of
pseudopodia
• Found in water sources throughout the world
• Few cause disease
 Entamoeba histolytica

• Carried asymptomatically in the digestive tracts

of humans
• No animal reservoir exists
• Infection usually occurs by drinking water
contaminated with feces that contain cysts
• Trophozoites migrate to the large intestine
where they multiply
Entamoeba histolytica
• cosmopolitan distribution
• no animal reservoirs
• typical fecal-oral life cycle cyst
• inhabits large intestine
• facultative virulent pathogen
• estimated 50 million cases/year
• 100,000 deaths/year

Entamoeba dispar
•morphologically identical
•non-pathogenic
trophozoite
Pathogenesis of Amebiasis
• NON-INVASIVE
• ameba colony on intestinal mucosa
• asymptomatic cyst passer
• non-dysenteric diarrhea, abdominal
cramps, other GI symptoms
• INVASIVE
• necrosis of mucosa  ulcers, dysentery
• ulcer enlargement  dysentery, peritonitis
• metastasis  extraintestinal amebiasis
• cessation of cyst production
 Entamoeba histolytica
• Three types of amebiasis can result from infection
– Luminal amoebiasis
• Least severe form that is asymptomatic
– Invasive amoebic dysentery
• More common form of infection
• Characterized by bloody, mucus-containing stools
and pain
– Invasive extra-intestinal amoebiasis
• Trophozoites carried via the bloodstream throughout
the body
• Maintaining clean water is important in prevention
• ulcers with raised borders
• little inflammation between lesions
• ‘flasked-shaped ulcer’
• trophozoites at boundary of necrotic
and healthy tissue
• trophozoites ingesting host cells
• dysentery (blood and mucus in feces)
Ulcer Enlargement and Disease
Progression
• ameba expand laterally and
downward into lamina propria
• localized sloughing (ulcers coalesce)
• perforation of intestinal wall
• peritonitis
• 2o bacterial infections
• local abscesses
• ameboma (=amebic granuloma)
ameboma = inflammatory thickening
of intestinal wall around the abscess
(can be confused with tumor)
The Course of Amoebiasis Due to Entamoeba histolytica
Extraintestinal Amebiasis
• metastasis via blood stream
• primarily liver (portal vein)
• other sites less frequent
• ameba-free stools common
• high antibody titers
Amebic Liver Abscess
• chocolate-colored ‘pus’
• necrotic material
• usually bacteria free
• lesions expand and
coalesce
• further metastasis, direct
extension or fistula
 Pulmonary Amebiasis
• rarely primary
• rupture of liver abscess
through diaphragm
• 2o bacterial infections
common
• fever, cough, dyspnea,
pain, vomiting

Cutaneous Amebiasis
• intestinal or hepatic fistula
• mucosa bathed in fluids
containing trophozoites
• perianal ulcers
• urogenital (eg, labia,
vagina, penis)
 Acanthamoeba and Naegleria
• Cause rare and usually fatal brain infections
• Common inhabitants of natural waterways as well as
artificial water systems
• Contact lenses users who use tap water to wash their
lenses can become infected
• Acanthamoeba diseases
– Infection occurs through cuts or scrapes, the
conjunctiva, or through inhalation
– Acanthamoeba keratitis results from conjunctival
inoculation
– Amebic encephalitis is the more common disease
 Acanthamoeba and Naegleria
• Naegleria disease
– Infection occurs when swimmers inhale contaminated
water
– Amoebic meningoencephalitis results when
trophozoites migrate to the brain
• Prevention is difficult because these organisms are
environmentally hardy