MICROBES AND

DISEASE

Dr Nagamani bora
  Koch’s postulates & how these are
modified to include our knowledge of
molecular genetics

LEARNI  Definition of different classes of

NG pathogen

OUTCO  Routes of infection with examples

MES  Definitions of Food borne infection and

intoxication
 ONLY Bacterial examples of each of these
 Epidemiology of food borne disease
A BIT OF HISTORY- SIGNIFICANT
MILESTONES
1680s
 Anton van Leeuwenhoek (inventor of the microscope) discovers
`animalcules'
 FIRST DEMONSTRATION OF THE POWER OF *

EPIDEMIOLOGY

 Epidemiology: Occurrence-Cause-Prevention
1854
 Cholera epidemic in Soho area of London traced to
contaminated water by Dr John Snow
 *
CHOLERA TODAY
IN THE NEWS…HAITI IN THE NEWS XMAS
2010/11
 Outbreak began October 2010 in an area not affected by
earthquakes but then spread to refugee camps
 Dec 7th 2010: 2,071 deaths and 88, 789 infected since the
outbreak of the disease.

Unsanitary living conditions, limited medical support and

local superstitions adding to problem.
 Molecular typing showed that the outbreak strain came
from staff in the UN aid camp.
 *
CHOLERA TODAY
– NOT IN THE NEWS…

South Sudan,
May-
Angola, 2013 OCTOBER
Outbreak in 2014 – Rainy
slums season
Sierra Leone
2012 - Caused by
a combination of
heavy rain,
Congo 2011 overcrowding,
poor sanitation
and dirty water

Zimbabwe 2008/9- No clean

water
2017 ……
 PAY ATTENTION

Big Questions
What, How and Where?
 *
SIGNIFICANT MILESTONES:
MAJOR CHANGE IN UNDERSTANDING

1870s
 Louis Pasteur and John Tyndall postulate that
there is no spontaneous generation of life -
microbial life comes from existing life.
 Pasteur shows that life can exist without air
 when asked to investigate why French beer is
inferior to German beer, discovers incidentally
that strictly anaerobic life can exist!
 *
SIGNIFICANT MILESTONES:
MAJOR CHANGE IN UNDERSTANDING

1881
 Robert Koch adds gelatin to liquid media forming a
gel; uses this to grow cultures.
 Laboratory culture methods (esp. sterile technique)
developed sufficiently to allow study of pure
cultures
 German bacteriologist

Julius R. Petri worked with Koch

 http://www.giantmicrobes.com/
SIGNIFICANT MILESTONES:
MAJOR CHANGE IN UNDERSTANDING

1884
 Robert Koch puts forward his findings on
tuberculosis.
 Described criteria needed to prove a specific
microbe caused a specific disease
 “Koch’s Postulates”
1. A specific microorganism can always be
found associated with a given disease
“Koch’s Postulates”
2. The microorganism can be isolated and
grown in pure culture in the laboratory
(not always true: e.g. Viruses or M. leprae)

“Koch’s Postulates”
3. The pure culture of the microorganism
will produce disease when inoculated into a
susceptible animal (host)

“Koch’s Postulates”
4. It is possible to recover the microorganism
from the experimentally infected host

“Koch’s Postulates”
SIGNIFICANT MILESTONES *
MAJOR CHANGE IN UNDERSTANDING

Vector borne diseases - 1900

 Walter Reed discovers that Yellow fever is
transmitted by mosquitoes rather than by direct
contact

 devastating viral disease that can cause bleeding from the

eyes, nostrils, anus and other mucous membranes
 Initiated study of zoonotic disease
 *
SIGNIFICANT MILESTONES
MAJOR CHANGE TO TREATMENT
1928
• Alexander Fleming notices that the mould Penicillium notatum kills
the bacterium Staphylococcus aureus.

– He grows the mould and discovers that its extracts also kill the bacterium (Nobel
Prize, Medicine, 1945).

• cultures kept for 2 to 3 weeks before discarding them

• culture left on a lab bench and went on vacation
• While away, the culture was subjected to a cold spell and then a hot spell of weather

The only conditions under which the

discovery could have been made!

 *
SIGNIFICANT MILESTONES
MAJOR CHANGE TO TREATMENT
1935
 Gerhard Domagk synthesizes Protonsil (sulfanilamide), used to
kill Streptococcus.
 He uses large doses on his own daughter as she lies
dying; she makes a dramatic recovery
 Nobel Prize, Medicine, 1939

1944
 Waksman et al. discover streptomycin for treatment of TB
 Nobel Prize, Medicine, 1952
SIGNIFICANT
MILESTONES: *
SUCCESSFUL
APPLICATION
OF KNOWLEDGE
1979
 Smallpox is declared to be eliminated.
 The only microbial disease ever completely
defeated!
 Jenner discovered vaccination using cowpox in 1796 but
had no understanding of what caused the disease
 *
SIGNIFICANT MILESTONES
MAJOR CHANGE IN UNDERSTANDING

1982
 Stanley Prusiner describes a class of infectious proteins
called Prions
 cause scrapie in sheep and now BSE in cows (mad cow
disease) (Nobel Prize, Medicine, 1997)

1983
 Luc Montaigner and Robert Gallo report that the virus they
have discovered (HIV) probably causes AIDS.
 Retroviruses not described until 1970’s; identified as cancer-
causing viruses (LM Nobel Prize, Medicine, 2008 )
 *
SIGNIFICANT MILESTONES
2012 total: 6632
IMPACT OF GENOMICS 2013 total; 19,078
2015 total; 56,058

1995
• Craig Venter et al. determine the first complete sequence: that
of the bacterial pathogen Haemophilus influenza.
1996
• Complete DNA sequence of all the chromosomes of a
eukaryote (S. cerevisiae) determined.
2003
• Complete genome of 128 eubacteria, 34 archaea, 1174 viral
genomes
2015
• 56,058 prokaryotic, 5048 viral genomes & 2556 Eukaryotes
sequenced (including human genome)
KOCH’S MOLECULAR
POSTULATES
(1) A gene found associated with a pathogen
(2) mutation of that gene attenuates the
virulence of a pathogen
(3) transfer of that gene confers pathogenicity
on another organism
 *
Factors Causing Emerging
Infections
TYPES OF PATHOGENS

(a) pathogen
e.g. Salmonella typhi
(b) opportunistic pathogen
e.g. Listeria monocytogenes
(c) obligate pathogen
e.g. Chlamydia trachomatis
 *
Emerging Infections Worldwide

Numbers indicate the people estimated to be living with

HIV/AIDS at the end of 2002. Locations of recent outbreaks of
emerging infectious diseases are indicated. Source: WHO
 *
Importance of Infections to Human Health

(a) Developed Countries

 *
Importance of Infections to Human Health
(b) Developing
Countries
FOOD BORNE ILLNESS:
DEFINITIONS
 FOOD POISONING results from ingestion of
toxins present in food - toxins interfere with
biochemical processes
 FOOD-BORNE INFECTION results from the
ingestion & growth of pathogenic microbes -
mostly caused by improper storage/handling or
inadequate cooking of food
 Food Borne illnesses

Food Borne Infections

Food Poisoning
Bacteria, Viruses, protozoa

Toxins
Fungal and some bacteria

Fungal toxins Endotoxins

LPS
Bacterial toxins
Exotoxins
SYMPTOMS
 Uncomfortable life-threatening

 Dysfunction of the intestinal

tract (common)
Cholera bed

 other organs are affected (more rarely)

 e.g. E. coli O157:H7 (kidneys)
Listeria monocytogenes (multiple organs)
EPIDEMIOLOGY (PATTERNS
OF DISEASE)
 Sporadic (individual cases, unknown sources)

 Family outbreak (home prepared foods)

 General outbreak (distributed foods)

CAUSAL AGENTS
 Bacteria: infections & intoxications
 viruses: infections (Dr Mellits’ lecture)
 fungi: mostly intoxications
 protozoa: infections
 e.g. amoebic dysentery
 helminths: infections
 e.g. tapeworm

Taenia solium
 *

TOXINS
 may be environmental chemicals or produced naturally
e.g. Rhubarb leaves - contain high levels of oxalic
acid
Kernels of soft fruit - contain hydrogen
cyanide (1981; 24 children poisoned in Israel)
Polar bear liver - toxic levels of Vitamin A
Gills of shell fish (Dead Man’s Fingers) -
contain potent toxin
 *

TOXINS
• Chemical toxins
e.g. From packaging - 1981:
tin & lead poisoning following consumption of canned grapefruit;
acidic contents leached metals from can
ACCIDENTAL
Contamination - 1990:
Spanish olive oil contaminated with industrial oils “marked” with
aniline dyes to prevent consumption; caused blindness & death
DELIBERATE
 *

MICROBIAL TOXINS
• fungal toxins (mycotoxins)
e.g. Claviceps purpurea - purple coloured fungus
grown on cereal crops and produces potent
toxin (ergot); associated with breads

Aspergillus flavus species produce spores

containing a toxin (aflatoxin): associated with
peanuts
BACTERIAL TOXINS
• EXOTOXINS: extracellular diffusible toxins; normally
proteins secreted during exponential growth

• Different modes of action identified

– membrane damaging (e.g. C. perfringens)
– Inhibit protein synthesis (e.g. E. coli)
– Activate host cell signals (e.g. V. cholera)
– Activate immune response (e.g. S. aureus)
– Proteases (e.g. B. anthracis)

* No need to learn these examples

BACTERIAL TOXINS
 Bacillus cereus
 spores are heat resistant & germinate in cooled food
after cooking.
 Vegetative cells produce heat stable toxin during growth
which survives in reheated food
- associated with cereals & rice
- causes vomiting OR diarrhoea & abdominal
cramps
- symptoms last only 12 hours
BACTERIAL TOXINS
• Clostridium botulinum
• Spores are heat resistant & germinate in cooled food
• Vegetative cells produce potent neurotoxin during
growth, but toxin is heat sensitive

– associated with meat dishes or canned goods

(anaerobe) not reheated before eating
– onset of symptoms within 12-36 h; death within
24 h (without antidote)
– symptoms include headache, dizziness, diarrhoea,
muscle paralysis, blurred vision
BACTERIAL TOXINS
• Clostridium perfringens
• natural part of animal gut microflora (anaerobe) &
spores found in faeces
• heating induces spore germination & removes oxygen
• Vegetative cells rapidly produce toxin during growth
– associated with cooked meat products (pies, stew, large
joints)
– onset of symptoms within 8-22 h; last for 12-24 h
– abdominal pain, diarrhoea & prostration
BACTERIAL TOXINS
 Staphylococcus aureus
 natural part of skin & nasopharynx microflora &
transmitted to food by poor hygiene.
 Heat stable toxin produced, not destroyed on heating.
BUT 106 cells needed to cause food poisoning
- associated with salad bars & prepared foods
- onset of symptoms within 1-6 h of ingestion;
last for 24 h
- nausea, cramps, diarrhoea & prostration
FOOD BORNE ILLNESS
 FOOD POISONING results in biochemical
dysfunction (often life-threateneing) but
generally NOT a raised temperature
 FOOD-BORNE INFECTION generally
results in symptoms of infection (raised
temperature, muscle soreness) due to
immune response.
BACTERIAL TOXINS
 Some of symptoms of bacterial infection
caused by natural toxicity of cell
components

 ENDOTOXINS: parts of the structural

components of the bacterial cell structure
 e.g. LPS of Gram-negative bacteria
 can be released as bacterial cells are destroyed by
host immune system or by antibiotic treatments
BACTERIAL INFECTIONS
• Salmonella
• normal habitat intestinal tract of mammals, birds & reptiles;
• transmitted by faecal contamination. Infective dose = 10 -
106
- many sources (pets, poultry, eggs, raw meat)
- onset of symptoms within 6-48 h of ingestion;
last for 1-8 d; rarely fatal
- abdominal pain, diarrhoea, prostration,
frequent vomiting, fever
- ambulant cases continue to excrete for many
weeks (“Typhoid Mary” early 1900’s)
BACTERIAL INFECTIONS
 Campylobacter
 normal habitat intestinal tract of birds;
contamination of poultry carcasses common.
- mainly from poultry or other raw meat
- begins with flu-like symptoms, fever &
headache, followed by profuse diarrhoeas,
abdominal cramps but no vomiting
- recovery can take 1-8 weeks
BACTERIAL INFECTIONS
 Listeria
 normal habitat intestinal tract of mammals, soil
& plants
 psychrotrophic organism
 can grow at refrigeration temperatures
 many sources (dairy, salads, cold meats)
 long incubation period; cell numbers reach critical
threshold during cold storage
 mild flu or gastroenteritis to meningitis, also
abortion & still birth
 opportunistic pathogen, only infects immuno-
compromised (e.g. pregnant women)
 Cases of infection are Rare but have 30% mortality rate
PREVENTING FOOD BORNE
ILLNESS
 Simple hygiene rules can dramatically reduce risk
 hand washing reduces faecal-oral transfer
 correct storage to prevent microbial growth
 separation of raw & cooked products to prevent
cross-contamination
 correct cooking of foods
…YOU DO NOT NEED TO
KNOW
 Introductory material on relevance of microbiology
& subjects studied at SB
 Any Dates/evolutionary time scales
 Examples given as “exceptions to the rule”
 Any phylogenetic trees
 Names of Archaea, protozoa, algae or fungi