Bacillus cereus

‘Bacillus’
 Derived from the latin word ‘bacillum’ meaning staff or walking stick; it is used to group
rod-shaped bacteria
‘cereus’
 From the latin word ‘cerea’ meaning waxy, it refers to the appearance of colonies grown
on blood agar.

General Characteristics of Bacillus cereus

 Gram-positive rods
 Motile
 Facultatively anaerobic
 Spore-forming
 Toxin-producing
 Catalase: Positive
 Oxidase: Positive
 Appears as large, feathery, dull, gray, granular, spreading colonies on 5% sheep
blood agar media
 
Bacillus cereus spores

 B. cereus is known to have the ability to withstand time and harsh environments due to
these spores that are resistant to heat, dehydration and other physical stresses.
 Frequently isolated from soil, which germinates and grows when brought in contact with
organic matter or an insect or animal host.
 Hydrophobic spores that adhere to surfaces, which makes it hard to remove by cleaning
or sanitation
 Has appendages and/or pili which contributes to its adhesion

Transmission (How do you get it?)

- The primary mode of transmission of B. cereus is the ingestion of contaminated food.

B. cereus is not a contagious pathogen, therefore it cannot be spread from person to
person.
- Epidemiology: Incorrect food preparation, particularly temperature abuse, poor hygiene
during canning or inadequate reheating (especially in congregate setting such as
restaurants and schools) pose the greatest risk of causing illness. Outbreaks tend to
have high attack rates (in excess of 50%).
 - Exposure‐prone groups: Those exposed to contaminated foods, food handlers,
residents in residential institutions.

Occurrence in Food

- As B. cereus is usually found in soil, raw plant foods such as rice, potatoes, peas, beans
and spices are common sources of B. cereus
- Meats, milk, vegetables, and fish are also possible sources
- The presence of B. cereus in processed foods results from contamination of raw
materials and the subsequent resistance of spores to thermal and other manufacturing
processes

Serotypes/Pathotypes

A provisional serotyping scheme for Bacillus cereus has been developed

Serotypes 2, 6, 8, 9 and 10 – Strains from incidents of food poisoning characterized by

a longer incubation period and with diarrhea as the main symptom

Serotypes 1, 3, 5 - Strains from incidents of rice-associated emetic illnesses

Diseases or Infections Caused and Symptoms

B. cereus intoxication produces two types of foodborne illness — the emetic (vomiting)
and diarrheal syndromes. These two illnesses are caused by two distinct metabolites.

1. Emetic syndrome
- The emetic syndrome is an intoxication that is caused by the ingestion of a heat-
stable toxin called cereulide that is pre-formed in the food during growth by B.
cereus. This syndrome has a short incubation period and recovery time
- Symptoms include nausea, vomiting and abdominal cramping, which occur within
1–5 hours of ingestion
- Recovery is usually within 6–24 hours
- Emetic syndrome resembles S. aureus food poisoning in its symptoms and
incubation period.
- It is commonly associated with fried rice
 B. cereus is frequently present in uncooked rice, and heat-resistant spores may
survive cooking. If cooked rice is held at room temperature, the vegetative
forms multiply, and cereulide is produced. Since this is a heat-stable toxin, it
can survive brief heating, such as stir frying.

2. Diarrheal syndrome
- The diarrheal syndrome is an infection caused by heat-labile enterotoxins
produced by B. cereus inside the host
 - The incubation period before onset of disease is 8–16 hours and the illness usually
lasts for 12–14 hours, although it can continue for several days
- Symptoms are usually mild with abdominal cramps, watery diarrhea and nausea
- Diarrheal syndrome resembles C. perfringens food poisoning
- It is commonly associated with meat and vegetables

Toxins Produced

Bacillus cereus produces one emetic toxin and three different enterotoxins

Emetic toxin (cereulide)

- Cereulide is produced by a non-ribosomal peptide synthetase (NRPS) complex. The

entire NRPS cluster has been characterised resulting in a highly specific method for
detection of cereulide-producing B. cereus strains.
- The emetic toxin is highly resistant to environmental factors, showing stability from
pH 2 to 11 and during heating to 100°C for 150 minutes. Cereulide toxin is not
inactivated by re‐heating food.

Enterotoxins

- The three types of enterotoxins produced by B. cereus are:

 the three component enterotoxin haemolysin BL (HBL)
 the three component non-haemolytic enterotoxin (NHE); and
 the single component enterotoxin cytotoxin K (EntK).
- Two out of the three (HBL and NHE) are associated with the diarrheal form of the
disease. All three enterotoxins are cytotoxic and cell membrane active toxins that
will make holes or channels in membranes.
- After consumption of food containing B. cereus, the enterotoxins are released into
the small intestine during vegetative growth following spore germination, and by any
surviving vegetative cells.
- The diarrheal enterotoxins can be produced in the temperature range of 10–43°C,
with an optimum of 32°C, and at pH levels between pH 5.5–10, with an optimum of
pH 8. The diarrheal enterotoxins are stable at pH 4–11 and inactivated by heating to
56°C for 5 minutes.
- Up to 26% of B. cereus vegetative cells can survive conditions that simulate
passage through the stomach. The survival rate of the vegetative cells is dependent
on the strain type, phase of vegetative cell growth and the gastric pH.
 Additional notes about toxins produced:

 Diarrheal enterotoxins are unstable at low pH and are degraded by digestive

enzymes, therefore any enterotoxins pre-formed in food would be destroyed during
passage through the stomach and would not cause illness if ingested.
 In contrast, spores of B. cereus are able to pass unaffected through the gastric
barrier. The spores contain receptors that need triggering by certain low molecular
weight substances to commence germination. These inducers may be present in the
food as well as the intestinal epithelial cells. The spores germinate, grow and
produce enterotoxins in the small intestine.
 A crucial virulence factor required for causing the diarrheal symptoms is the ability of
the vegetative cells and spores of B. cereus to adhere to the epithelial cell wall of
the small intestine.

Implications/importance in the food industry

Knowledge about the genetic aspects of Bacillus cereus and their complex mechanisms
for toxin production highlights the importance of the control on food and dairy industries
and the development of new strategies for control.

Prevention

Because B. cereus endospores are extremely heat resistant, they are likely to survive
cooking at temperatures that would otherwise destroy foodborne pathogen cells. Heat
resistance increases with increasing salinity and decreases with increasing acidity.

According to the National Institutes of Health (NIH), the National Institute of Allergy and
Infectious Diseases (NIAID), the National Food Processors Association (NFPA), and the FDA
Food Code 2013, the suggestions stated below are good methods and practices that destroy B.
cereus:

1. Steaming under pressure, roasting, frying, and grilling foods will destroy the
vegetative cells and spores if temperatures within foods are ≥ 63ºC
2. Heating to ≥ 63ºC and reheating to 74ºC for 15 seconds will destroy the vegetative
cells. Once cooked, the rapid cooling of the product will prevent any spores present
from germinating. However, it is important to note that foods cannot be made safe to
eat if toxins have already been formed.
3. Keep hot foods above 57ºC and cold foods below 5ºC to prevent the formation of
spores.
4. Refrigerate leftovers properly by cooling rapidly to ≤ 5ºC. Make sure leftover
containers are not stacked close together; this will allow for adequate airflow around
the food and ensure rapid cooling.
NOTE: Since foods containing the emetic toxin need to be heated to 121ºC for more than 80
minutes, reheating foods until they are steaming is not enough to inactivate the emetic toxin and
efforts should be devoted to prevention.

Answers to Guide Questions

1. Compare and contrast the emetic and diarrheal syndromes of B. cereus poisoning. What
food vehicles are closely associated with the two types?

Emetic syndrome Diarrheal syndrome

Type of Foodborne Illness Intoxication Infection
Metabolite Cereulide (heat-stable) Enterotoxins (heat-labile)
Incubation Period Short (1-5 hours) Long (8-16 hours)
Nausea Abdominal cramps
Symptoms Vomiting Watery diarrhea
Abdominal cramps Nausea
Meat
Food Vehicle Fried rice
Vegetables

2. Discuss the importance of endospore formation in the survival and virulence of B.cereus.
- As mentioned earlier, the spores produced by Bacillus cereus are hydrophobic,
which makes the spores hard to remove by cleaning and sanitation. The spores also
adhere to surfaces; they have appendages and/or pili which contributes to its
adhesion.