Eukaryotes in Food borne disease:

Protozoa, parasites and fungi

 Protozoa and Parasites
only cause foodborne infections
 FOOD BORNE PATHOGENS
Tree of life

(16S/18S rDNA
comparisons)

•Enterobacteria
• Gram posi0ve • Protozoa

• Parasi0c
worms

• Fungi
 EUKARYOTIC AND BACTERIAL CELLS

Bacteria

• Smaller (0.2-5 μm)

• No nucleus
• Larger in size (10-100 μm) • No self-contained organelles
• Usually only have cell membrane • Sensi0ve to an0bio0cs
• True nucleus with condensed DNA
in chromosomes
• Organelles: mitochondria, Golgi
apparatus, ER, vacuoles
 PROTOZOA AND PARASITIC WORMS
• Eukaryotic pathogens p. 42

• Cause infections

• Protozoa - unicellular microorganisms

• Worms - multicellular organisms

• Complex life cycles which require a host

• Some are transmitted human to human, others involve

animals

• The resting stage initiates the disease (cysts, oocysts, eggs)

 PROTOZOA: BASIC LIFE CYCLE

Encystment:
Trophozoites to cysts

Excystment:
Cysts to trophozoites

Asexual propagation
Trophozoites: Vegetative cells
Cysts: Dormant stage

Sexual reproduction
Trophozoites mature to gametes pairs which form zygotes.
Result: Sporozoites encased in Oocysts
Entamoeba histolytica : AMOEBIC DYSENTERY

Host: humans only

(Transmitted through the fecal-oral route)
Entamoeba histolytica

AMOEBIC DYSENTERY

Bloody diarrhea

Liver infections

It is hard to eliminate: asymptomatic carriers

Control: Good sanitation

More common in developing countries (100,000 deaths annually)
Toxoplasma gondii and TOXOPLASMOSIS
Host: humans, animals (cats – required!)
 Toxoplasma gondii
TOXOPLASMOSIS

Mainly asymptomatic (no disease) in healthy people

Serious gastroenteritis in immunocompromised people

May cause neurological birth defects

Transmission:
Soil on food contaminated with oocysts
Undercooked meat containing cysts
Giardia lamblia (Giardia intestinalis)
GIARDIASIS

2 stage life-cycle:
Trophozoite
Cyst (infectious)

Dose: low (10-110 cysts)

Incubation: 1-2 weeks
Duration: 5 days or longer

Symptoms: diarrhea, gas, weight loss

Giardia lamblia (Giardia intestinalis)
GIARDIASIS
Host: Humans and animals
Asymptomatic in some people and most animals

Mostly associated with untreated drinking water

“Beaver fever”

Survives a long time (>100 days)

Control: chlorinated drinking water

Cysts are now known to survive
Can be removed by filtration
Cryptosporidium parvum & CRYPTOSPORIDIOSIS

Sexual reproduction life-cycle:

Oocyte excyst and release sporozoites in the intestine

Dose: Low (<10 oocysts)

Incubation: 2 – 10 days
Duration: 1-2 weeks (long)

Symptoms: diarrhea, abdominal pain, and vomiting

Immunocompromised: invade lungs
Cryptosporidium parvum & CRYPTOSPORIDIOSIS

Host: Humans and animals

Parasitic for several animals, especially calves

Resistant to chlorination procedures

small oocysts (difficult to filter (<10 mm required))

Epidemics traced to water supplies

1993 Milwaukee outbreak:

Contamination of a water purification plant
>405,000 infected and 100 deaths
 Control of Giardia and Cryptosporidium
Alternative water treatments:
ozonization
UV
photocatalytic disinfection
heat (>60 C for 5 min)

low pH
dessication
 PARASITIC WORMS CARRIED IN FOOD

p. 46

Similar life cycle to protozoa

The dormant stage is called EGG** or CYSTS **

(**cause infection)

Larvae develop into worms which then mate and form

zygotes
 Trichinella spiralis & TRICHINOSIS

Larvae migrate into the lymph and blood, and encyst in muscles

The disease contracted only by eating a meat-eating animal

(pigs, bears). Cysts are not present in the environment.
Trichinella spiralis & TRICHINOSIS
Host: Humans and Animals

After eating meat containing live cysts:

Mild first symptoms are mild: pain and mild
diarrhea

Weeks later:
fever, weakness and muscle pain.

Control:
Prevent animal infection
Cook pork to 160 F
Freezing might be effective (< 15 F)
Taenia saginata BEEF TAPEWORM
Taenia solium PORK TAPEWORM

•
Taenia saginata BEEF TAPEWORM
Taenia solium PORK TAPEWORM
Usually the infection is limited to the intestines

Symptoms are subtle:

weakness
nutritional deficiencies or susceptibility

Head attaches to intestine, and worms can

grow to 4 – 12 meters long

T. solium can cause brain infection when

transmitted through the fecal-oral route
(human-human)
Diphyllobothrium latum FISH TAPEWORM

Ingestion of undercooked, raw fish

We are unsuitable hosts – parasites do

not develop further or reproduce

Fish that is thoroughly cooked,

brined, or frozen at -10°C for
24–48 hours can be consumed
without risk of D. latum infection

Majority of infections in the US associated with dishes

prepared at home
 Review:
Entamoeba histolytica - Amoebic dysentery
Toxoplasma gondii – Toxoplasmosis
Giardia lamblia - Giardiasis
Cryptosporidium parvum - Cryptosporidiosis
Trichinella spiralis – Trichinosis
Taenia saginata – Beef Tapeworm
Taenia solium – Pork Tapeworm
Diphyllobothrium latum – Fish Tapeworm

Parasitic worms and protozoa all cause food borne infections

Contracted either by poor sanitation or eating contaminated meat

The diseases are initiated by eating/drinking Cysts/Eggs/Oocysts

 Review:
Parasitic worms and protozoa

Have a basic understanding of the life cycle:

- how it is transmitted (e.g. water, muscle, soil)
- human only or animal-human
- unique features: e.g. cat and Toxoplasma
- organs affected by illness
 Mycology

Mycology (Myco = fungus) (-ology = study)

General Features of the Fungi

Eukaryotes – possess true nuclei and other membrane-bound

organelles

Possess cell walls containing chitin

Heterotrophic metabolism - they must consume preformed

organic matter (Non-photosynthetic)
 Reproduction can occur by a sexual or asexual process

Asexual and sexual reproduction of fungi

The sexual state is referred to as the teleomorph
The asexual state of a fungus is termed the anamorph
Many fungi exhibit both types of reproduction the holomorph

Both forms of reproduction can result in spores

Slightly more resistant to environmental stress than vegetative cells (not nearly as
resistant as bacterial endospores)

Generally designed for dispersal

Germinate to form a vegetative cell when conditions are right

 Mold form one common group of fungi

Molds

The vegetative growth of molds

consists many thread-like, walled,
multi cellular, cylindrical hyphae
A single filament is called hypha

Hyphae usually grow together

across a surface and form
compact tufts, collectively
called a mycelium

Cells within a hypha are usually

separated by septum (some molds
may not have septa)
 The life cycle of a mold

Asexual vegetative growth -occurs at the ends of hyphae, and occasionally at

branch points

conidia
Asexual growth with spore formation:

The asexual lifecycle of Penicillium species

sporangia
The life cycle of Rhizopus species: An example of a holomorph
 Yeast are another group of fungi

Single cells

Circular or oval shape

Asexual reproduction is via budding (or occasionally

fission)

Yeast chains are called pseudomycelium

Sexual reproduction produces:
The spores reside inside an ascus
Commonly there are 4 ascospores (we will see this in the lab)
Fungal Toxins in Foods: Yeasts and Molds

Yeasts and molds (including mushrooms) p. 52

Yeasts live primarily as unicellular microbes

Molds form highly branched colonies: multicellular

 Yeasts and Molds

Important in food fermentations

Antibiotic production

Biopesticides

Edible (Some mushrooms can be poisonous)

Enzyme production

Spoilage microbes & Toxins (molds only)

 Msome molds cause foodborne
intoxications
Soil microbes
 Mold Structure

Mycelium
A mat-like colony or
microscopic branches
Hypha – one branch
of the mycelium

Spores
Large quantities are made
Stress tolerant
(less so than
bacterial endospores)
Ergot mycotoxin poisoning

Ergot: class of mold toxins which affect neurons

Fungal Agent: Claviceps purpurea

Plant pathogen (rye)

Grows inside rye and wheat

kernels when damp

Black kernels contain the toxin

Ergot Poisoning
Foodborne Intoxication

Heat-stable toxin (ie baked bread)

St. Anthony’s fire:

Neural degeneration (pain and convulsions)
Hallucinations, delusion, confusion.
Gangrene in extremities

Epidemics in the Middle Ages.

Highest incidence during starvation

Associated to witchcraft –
Salem’s trials
 Aflatoxin mycotoxin poisoning

Agent: Aspergillus flavus

Widespread mold that can grow in many grains, nuts, and cereals (warm
and damp)

Produces Aflatoxin before or after harvest

 Aflatoxin poisoning
Causes disease only at high
concentrations (0.01 g)

Example: visibly rotten food

Metabolized in the liver and causes liver Aflatoxin B1

damage and tumors

Most potent natural carcinogen: Intercalates into DNA

Low doses over many years may cause cancer

 Control

! keep grains, nuts, and cereals cool and

dry

! Sort and examine to remove moldy

grains

! Detoxify using oxidizing agents or

alkaline treatments which denature the
toxin
 Prions
They are actually protein toxin not related to microbes
but the pathology is similar to an infection
Transmissible Spongiform Encephalopathy

Progressive and fatal neurodegenerative disease

brain dysfunction (ataxia, confusion), coma and death

Relation to foods: BSE or “mad cow disease”

Transmissible Spongiform Encephalopathy
Human Animal
! Creutzfedlt-Jakob • Scrapie in sheep
disease (CJD) • Chronic wasting disease
! Gerstmann-Straussler- (CWD) in mule deer and
Scheinker disease elk
(GSS) • Transmissible mink
! Fatal familial insomnia encephalopathy (TME)
(FFI) • Bovine spongiform
! Kuru disease1 encephalopathy (BSE) 2
• Feline spongiform
Gene0c or transmiQed through consump0on of encephalopathy (FSE)
neural 0ssue from an infected individual
1.Presumed transmi/ed through consump5on of human brains
2. Maybe transmi/ed through consump5on of neural 5ssue from scrapie sheep or BSE ca/le
BSE: Bovine Spongiform Encephalopathy

Mad cow disease.

First found in England in the mid 1980s

Appeared shortly after use of animal by products as cattle feed

supplements became widespread

1989-Britain banned use of animal by products in cattle feed

 BSE and human disease
In the 1990s, a new CJD variant arose
called vCJD

Affected young people and

correlated with beef consumption

Maybe more infected: long incubation

The infection responsible for the disease in cows is believed

to be the same one responsible for vCJD in humans
Proteinaceous infectious particles

1982, Stanley Prusiner at UCSF

Published the prion concept

Nobel prize in 1997

Hypothesized that the disease agent for TSE was a

protein which could direct the formation of more
proteins like itself

Normal proteins of animal tissues mis-fold and

become infectious
 How it works
Physical interaction of PrPc with PrPSc converts it into PrPSc

PrPSc comes from

a) Sporadic misfolding of PrPc

b) Introduction of foreign PrPSc particle from food

 BSE and human disease (vCJD)

Prions are highly resistant to heat and chemical treatments

Control:
Do not eat (bovine) meat
infected with BSE