+

Bacterial Food Poisoning

(infective)

Dr. Sarita Mohapatra

+
Learning Objectives

 Pathogen causing infective food poisoning

 Pathogenesis
 Nontyphoidal Salmonella
 Shigella spp.
 EHEC, EIEC
 Listeria monocytogens

 Lab diagnosis
+ Sterilization
Boiling 100

90

Bacteria die if heated

Pasteurization 80
long enough
70
Hot storage
60
Thermophilic
50

40
Bacteria
multiply Bacteria Bacteria
Mesophilic 30 rapidly multiply multiply
slowly
20
Psychrophilic
10

Chill storage Most bacteria cease to

Freezing 0
multiply but do
-10 not die

-18

Cold storage
+
Improper cooking Poor personal
or reheating hygiene

Chemicals
in food Cross- contamination

Insects and Improper food

rodents Food is not sterile handling

Infected food Unclean equipment

handlers

Use of unsound, Unsafe holding

unwholesome or temperatures that
unfit food permit bacterial growth
+ Definitions
 Food borne Infection
Ingestion of live organisms which subsequently invade
the host
 Food borne disease
Any disease caused or transmitted by food
 Food borne Intoxication
Ingestion of a pre-formed toxin
 Miscellaneous (allergies)
Consumption of a specific compound which triggers an
immune response
+
Food poisoning

Any acute manifestation of diarrhea or vomiting caused by

toxins produced by microorganisms called food poisoning

resulting from the ingestion of food or drink

 contaminated with living bacteria

 with toxin

 poison from plant and animals

+

WHO, 2016
+ Food poisoning caused by
bacterial infection
 Nontyphoidal Salmonella
 Enterohemorrhagic E.coli
 Enteroinvasive E.coli
 Shigella spp.
 Campylobacter jejuni
 Listeria spp.
 Yersinia enterocolitica
 Vibrio parahemolyticus
Food poisoning caused by
bacterial toxins
 Staphylococcus aureus
 Bacillus cereus
 Vibrio cholarae.
 Clostridium perfringenes
 Clostridium botulinum
 Enterotoxigenic E. coli
 +
Food-borne disease in India
Top 6 organism causing foodborne illnesses

 Salmonella sp (most common)

 E. coli

 Staphylococcus aureus

 Vibrio sp

 Yersinia enteroclitica

 Norwalk-like virus

National Centre for Disease Control, Government of

+ Secretory Inflammatory Invasive
Location Prox. SI Colon Distal SI
Type of illness Watery diarrhoea Dysentery Enteric fever

Stool No faecal leu Faecal leu Faecal leu

Mechanism ET/ bact. Adherence, CT/ Bact. Invasion, Bact. Penetrate
invasion mucosal damage, mucosa, evade RE
inflammation

Examples V. cholerae, ETEC, Shigella, Salm. Salm.

Cl. per, Staph. Typhi/para, V. Typhi/partyphi, Y.
aureus, B. cereus parahaemolyticaus, enterocolitica
Cl. difficile, C. jejuni
+
+
Relationship between type of food and pathogen
Food type Micro- organism

Meat or milk eaten raw or undercooked Nontyphoidal Salmonella, Campylobacter

Fish and other sea foods eaten raw or Vibrio spp

undercooked
Cooked food contaminated by handler Staph. aureus, less common Salmonella, Shigella

Cooked food held warm Clostridium perfringens, Bacillus cereus

Cooked food held for extended period at low Listeria monocytogenes, Yersinia enterocolitica
temperature
Cooked food held for extended periods under Clostridium botulinum
anaerobiosis
Retorted food under-processed Clostridium botulinum

Fermented food, incomplete fermentation Staphylococcus aureus, clostridium botulinum

 +
Major presenting Probable Microbes Incubation Probable food sources
symptom Period

Vomiting Staphylococcus aureus 1-6 hours Prepared food (salads, dairy, meat)

Clostridium perfringens 8-16 hours Meat, poultry, gravy

Bacillus cereus 1-6 hours Fried rice, meat

Norovirus 24-48 hours Shellfish, prepared food, salads, sandwiches, fruit

Watery diarrhea Clostridium perfringens 8-16 hours Meat, poultry, gravy

Enterotoxigenic Escherichia coli 1-3 days Feces- contaminated with food or water

Cyclospora cayetanensis 1-11 days berries, basil

Clostridium parvum 2-28 days Vegetables, fruit, unpasteurized milk, water

Inflammatory Vibrio parahemolyticus 2-48 hours Raw shellfish

diarrhea
Nontyphoidal Salmonella 1-3 days Eggs, poultry, meat, unpasteurized milk or juice, fresh produce
species
Shigella species 1-3 days Feces-contaminated food and water

Shiga toxin-producing 1-8 days Ground beef, unpasteurized milk, and juice, raw vegetables, water
Escherichia coli
Campylobacter species 2-5 days Poultry, unpasteurized milk, water
+
 Food-borne illnesses passive surveillance systems
+ (Tip of the iceberg )
CDC estimates each year 48
million people get sick 128,000
are hospitalized, and 3,000 die
 Most patients have mild symptoms of short duration,

do not seek medical care

 Those that do access clinical care will not have the laboratory test
performed to determine a specific etiology

 Of those who seek medical care in whom an etiologic diagnosis is confirmed

will not be reported to the surveillance system(s)

 WHO estimates that food-borne diseases are notified in only 10% of cases
in developed countries and 1% in developing countries

Source: WHO Estimate of the global burden of foodborne disease 2015

+
WHO Global estimate of foodborne disease(FBD) in 2010

 About 600 million foodborne illness and 42 lakh deaths

 40% of the FBD burden was among children under 5 yrs

 1 in 10 people fall ill every year due to eating contaminated food

 Global burden of FBD was 33 million DALYs in 2010

 Most frequent cause of FBD were norovirus and Campylobacter

 Non- typhoidal salmonella enterica caused 2.3 lakh deaths Source: WHO Estimate of the global burden of foodborne disease 2015
+
 Development of new and emerging pathogens
 changed agricultural production methods
 consumption habits
 modern harvesting, processing and packaging
technologies

 global trade and food hygiene

 bioterrorism emerges as a new threat to food safety.

+ Salmonella food poisoning

 Gram negative, member of Enterobacterales

 Non spore forming bacilli
 Grows aerobically and anaerobically
 Oxidase negative, catalase positive
 Ferment glucose with production of gas
 Reduce nitrate to nitrite

 Motile by peritrichous flagella

 Produce H2S on TSI agar
+ SALMONELLA SPECIES
(Zoonotic)

 Sources:
 Ingestion of contaminated food (poultry and meat, raw
milk and dairy products, juice and seafood)
 raw or undercooked eggs (egg shells contaminated
with chicken feeds/feaces)

 Non-typhoidal (S. Enteritidis, S. Typhimurium) – poultry,

eggs, meat
 103 – 104

 Incubation period: 18-24hrs

 Diarrhea, vomiting, fever, abdominal pain
+ Pathogenesis

Enters gut lumen


Inflammatory mediators and
recruits neutrophil

inflammation of the epithelium
and actin rearrangement,
disruption of tight junction

Inflow of water and electrolytes
+

 Lab diagnosis  Treatment

 Culture: stool (XLD/DCA)
 Uncomplicated: No antibiotics
 Outbreak: food
 Invasive case: Antibiotics needed

National Salmonella and Escherichia centre (NSEC): Central Research

Institute, Kasauli
+ SHIGELLA SPECIES
(over 30 types)

Fecal-oral route
 Infective dose: 10-102 bacilli
 Human beings are the only natural host
 Incubation period: usually 48hrs
 Dysentery: freq. loose stool with scanty feacal matter
containing blood & mucus
+ Pathogenesis
Site of infection: M cells in payer’s patch of large intestine
Virulence factors
1. Shiga toxin: stxA, stxB
 A sub unit (A1, A2): helps in the biological processing
 A1 portion affects the eukaryotic ribosome specially involved
in inhibition of protein synthesis
 B sub unit (5B): binding to ganglioside receptor
 primarily a toxin for the vascular endothelium
 accounts for dysentery & severity of infections

2. Enterotoxin: ShET-1 and -2

 Causes inflammation of epithelial cells
Genetic Basis of Invasion

Pathogenic strains of Shigella carry a large plasmid in the outer

membrane protein (Virulence marker antigens, VMA) involved in the
process of invasion
 Shiga-toxin inhibits protein
synthesis in int. mucosa

mucosal damage

bacilli may invade & damage

vessels of mucosa

Increased Fibrin deposition &

prothrombotic tendency
HUS

Entry and dissemination of Shigella in epithelial cells: basolateral transfer

+ Enterohemorrhagic E. coli (EHEC)

 Produce two toxins

 Shiga-like toxin producing E. coli (STEC)
 Vero cytotoxin-producing E. coli (VTEC)

 Outbreaks of infection for elderly and young children

 Severaloutbreaks for O157:H7 in USA and Canada,

rarely O26:H1, O111

 Source:
hamburger meat, unpasteurized apple juice,
unpasteurized milk
+
Virulence factor

 Cytotoxin Similar to Shiga toxin: VT1, VT2

 Vascularcells are primary target of Stx1 causing micro-

angiopathy of capillaries

 VTs are A1/B5 type toxin, transferred by lysogenic phage

A1: acts at ribosomal level, inhibits protein synthesis
B: binds glycolipid receptor of renal endothelium

 Enterohemolysin
+

Symptoms: (Asymptomatic carriage to HUS)

 Diarrhea

 Hemorrhagic colitis

 Hemolytic Uremic Syndrome (HUS)

 Thrombotic thrombocytopenic purpura (TTP)

(microangiopathic hemolytic anaemia, thrombocytopenic purpura,
neurologic abnormalities, fever, kidney disease)
+
Enteroinvasive Esch. coli

 Invade colonic epithelial cell (Large plasmid)

 Virtually identical to Shigella spp.
 Nonmotile, nonlactose fermenter, anaerogenic
 Waterydiarrhea, fever, colitis, pus cells in stool,
blood and mucus, and tenesmus
 Serotypes are : 028, 029, 0112 etc.
+ Vibrio food poisoning

 Gram negative, straight, curved, or comma shaped

 Motile by single polar sheathed flagellum

 Oxidase positive and reduce nitrate to nitrite

 Growth stimulated by Na+

 Natural habitat: river, surface and sea waters and

sediments, sewage and wide variety of sea food

 Three species associated with foodborne illness V. cholerae,

V. parahemolyticus, V. vulnificus
+
Food poisoning by vibrio
Features Vibrio parahaemolyticus Vibrio vulnificus

Incubation period 9-24 hrs 16- 48 hrs

Main symptoms Diarrhoea accompanied by Primary septicaemia

abdominal cramps nausea Diarrhoea less common (40%)
Other symptoms Dysentery Wound
Wound extraintestinal infection
eye and ear infection
Common food associated Fish and other sea foods Fish and other sea foods
(crabs, oyster etc)
Susceptibility Reduced stomach acidity Underlying medical conditions
Usually adults
+ CAMPYLOBACTER JEJUNI

 Motile, microaerophilic, GNB

 99% C. jejuni, 1% other.
 Affects the small intestine (Jejunum and ileum)
 The organism is invasive: infection, lymph node
enlargement, mucosa is inflamed and edematous
 C/F: fever, pain abdomen, diarrhoea
Sources
Contaminated water, raw milk, and raw or undercooked meat,
poultry or shellfish
500-800 bacilli are needed
+
Symptoms
 Diarrhea
 Usuallywatery and sticky
 Can contain blood and fecal leucocytes

 Fever

 Abdominal pain (more severe than Salmonella or Shigella

infection)

 Nausea and vomiting

 Headache

 Muscle pain
+
How Does Campylobacter affect individual
cells in the intestines?

 Produces a toxin called Cytolethal Distending Toxin

(CDT).

 CDT activity requires activation of three genes: cdtA,

cdtB, and cdtC.

 CdtB is nuclease that damages DNA and causes cell cycle

arrest.

 Causes cell death

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+
Are There Long-Term Effects?

 Guillain-BarreSyndrome
 Immune system attacks own nerves
 40% caused by C. jejuni infection

 Arthritis, Reiter’s Syndrome

 Hemolytic Uremic Syndrome

 Septicemia
 Leadingto infection of nearly any organ, including appendix,
abdominal cavity, heart, nervous system, etc.
+ LISTERIA MONOCYTOGENES

 Coccobacillary to Gram positive bacillus

 Tumbling motility at 20- 25˚C by peritrichate flagella

 Aerobic and microaerophilic

 Sources:
 Soft
cheese, raw milk, improperly processed ice
cream, raw leafy vegetables, meat, and poultry.
 + 38

Internalin A – Helps Bacteria

to enter Cells

lyse phagosome by LPS &

Phospholipase, listerolysin O

Phospholipase helps in cell-to-

cell spread
+
+
Laboratory Diagnosis

 Confirm the clinical diagnosis by isolation of causative

agent from human specimens

 Ensure proper identification of the disease

 Determine if the same causative agent is present in

implicated food sources :
 biotyping, serotyping, antimicrobial susceptibility profile, phage
typing, plasmid profile, pulsed field gel electrophoresis, PCR
 +
 Onset and duration

 Fever Abdominal pain

 Appearance of stool Severe
 Blood and mucus Muscle cramps
 Blood without leukocytes Bloating
 Bulky white stool
Appendicitis-like
Tenesmus
 Profuse rice-watery stool
Vomiting
 Frequency

Common source
Antibiotic use
Travel history
H/O immunocomproise / immunosupression
Presence and severity of dehydration
 Patient Food handler Environ.

bacterial stool vomitus urine blood stool nose hands food Food
prep.
environ.
salmonella C C C C C

Staphylococcus C C C C C,T
aureus
Campylobacter C C C C
jejuni
Clostridium C,T C,T T C,T
botulinum
Clostridium C,T C
perfringens
shigella C C C
Vibrio C C c
parahaemolyticu
s
Bacillus cereus C C C,T
Yersinia C S C C C
enterocolitica

C,culture; I,immune electron microscopy; P,PCR probe; S,serology; T,toxin

testing
 Patient Food handler Environ.
bacterial stool vomitus urine blood stool nose hands food Food
prep.
environ.
Vibrio cholerae C S C C C
01 and non01

Shiga toxin E . C,T S C C,T

coli

viral

Norwalk virus I,P S I,P P

C,culture; I,immune electron microscopy; P,PCR probe; S,serology; T,toxin testing

+

 Sample Collection
 early stages of disease (heavy load of pathogens)
 before starting antibiotics
 in a clean tight-fitting container
 Investigation of outbreak – samples from multiple ill people

 Sample processing
 Immediately (<2 hours)
 If late (>2 hours), inoculate into transport medium / chilled
+ Transport Media For Clinical Specimens

 Cary-Blair medium
 Salmonella, E.coli, Shigella, Vibrio,
Yersinia
Alkaline peptone water
 Stable, storage for future use
Venkatraman-Ramakrishnan (VR)
Reduced APW & Thioglycollate
 Amies medium broth - Campylobacter

 Stuart’s medium
 Buffered glycerol saline
 Mainly for shigella
 Others salmonella, E.coli
 Not suitable for Vibrio
+
Direct examination
 Macroscopic
 Colour
 Volume
 Consistency
 Presence of blood, mucus, frank pus

 Microscopic
 Faecal leucocyte count
 Presence of ≥3 leukocytes/ HPF in 4 or more fields
 Simple, rapid and useful test for shigellosis,
campylobacteriosis, salmonellosis, EIEC (>50 pus cells /
HPF suggestive of shigellosis)
 Sensitivity 60-70%
+
 O157:H7 do not ferment sorbitol (MacConkey agar with

Sorbitol)

 Other methods of detection

 Toxin assay
 Antigen detection by ELISA
 Detection of enzymes
 Detection of genes encoding toxins by PCR
Laboratory diagnosis of Salmonella
samples
Stool
Blood
 Specimen- faeces or suspected food stuff Suspected food

 Culture
 Enrichment media- after 6 hrs of incubation
subculture on selective media.
 Selenite F broth
 Tetrathionate broth
 Selective media- incubated at 370c for 24 hours
 MAC (NLF colony)
 DCA (NLF colony)
 XLD (red colonies with black center)
 Wilson and Blair medium (shiny black colonies)
+Laboratory diagnosis of Listeria
Samples
Microscopy CSF
Gram- staining: Gram positive rods Blood
Wet mount: tumbling motility Amniotic fluid
Placental tissue
Stool
Culture Suspected food
(100g)
Clinical specimen from sterile sites
 Tryptic soy agar with 5% sheep blood
 Blood on conventional blood culture bottle

Non sterile specimen, food samples

 Selective enrichment broth
 Selective agar for culture

Identification and confirmation

+Laboratory diagnosis of Vibrio
Microscopy Samples
Blood
Gram- staining: Gram negative rods
Stool
Wet mount: Darting motility Suspected food
(100g)

Culture
Clinical specimen from sterile sites
 Mac conkey agar
 Blood in conventional blood culture bottle

Non sterile specimen, food samples

 Selective enrichment broth
 Selective agar for culture

Identification and confirmation

+ Culture
 Selective enrichment broth
 Alkaline peptone water (1%NaCl, pH 8.5)

 Mac conkey agar

 Small colorless colonies

 TCBS agar (selective media)

Yellow colonies ( V. cholerae, V. fluvialis, V. alginolyticus)

Green colonies (V. parahaemolyticus, V. mimicus, V. vulnificus)

+ Prevention of food poisoning
1.Keep Clean
Wash hands before handling food and during food preparation
Wash hands after going to the toilet

2. Separate raw and cooked food

Separate raw meat, poultry and seafood from other foods
Store food in containers to avoid contact between raw and prepared
foods

3. Cook thoroughly
Cook food thoroughly, especially meat, poultry, eggs and seafood
Reheat cooked food thoroughly

4. Keep food at safe temperatures

Do not leave cooked food at room temperature for more than 2 hours
Do not store food too long even in the refrigerator

5. Use safe water and raw materials

Use safe water or treat it to make it safe
Wash fruits and vegetables properly if eaten raw