Members of the genus Bacillus bring about bacterial spoilage of bread known as

rope. This is of major economic to the baking industry. Ropiness which is the most
important spoilage of bread after moldiness occurs particularly in summer when the
climatic conditions favour growth of bacteria. It is mainly caused by Bacillus subtilis
but Bacillus licheniformis, Bacillus magaterium and Bacillus cereus have also been
associated with ropy bread. The incidence of wheat bread spoilage caused by Bacillus
has increased during the last few years presumably because more bread is produced
without preservatives and often raw materials such as bran and seeds are added.
Spoilage of bread by rope formation may constitute a health risk, high numbers of
Bacillus subtilis and Bacillus licheniformis in foods may cause a mild form of food
illness. Consumption of ropy bread has been association with food-borne illness in
reports from Canada and the United Kingdom.

1. Bacillus licheniformis (Spoilage, especially causing ropiness in bread)

Bacillus licheniformis commonly found in soil, plant materials, and various food
production environments. This bacterium is known for its resilience, particularly its
ability to form heat-resistant spores, allowing it to survive harsh food processing
conditions, such as the high temperatures involved in baking. In bakery
products, Bacillus licheniformis is a significant spoilage organism, mainly due to its
production of extracellular enzymes that degrade starch and proteins. This degradation
can lead to undesirable changes in the texture and quality of baked goods, such as the
formation of a slimy or sticky crumb, commonly referred to as "rope spoilage." This
spoilage occurs when the bacterium produces enzymes that break down the starch and
proteins in the bread or other baked products, leading to a ropy, stringy texture, often
accompanied by off-flavors and odors.
Contamination can occur through raw materials, such as flour, or during handling,
storage, and packaging. If the products are improperly stored at warm
temperatures, Bacillus licheniformis can grow and produce spores, even in finished
products. Prevention strategies, including proper storage, hygiene, and packaging
practices, are essential to mitigate the risk of spoilage caused by this bacterium.
1.1. Microscopic Morphology
  Cell Shape and Structure:
- Rod-shaped (bacillus): Bacillus licheniformis cells are rod-shaped, cylindrical
bacteria.
- Size: The bacterial cells typically measure about 0.6 to 0.8 µm in diameter and
1.5 to 3.0 µm in length.

Figure 3.1. Bacillus licheniformis

 Gram staining: Bacillus licheniformis is a Gram-positive bacterium, meaning it
retains the crystal violet stain during the Gram-staining process due to the thick
layer of peptidoglycan in its cell wall.
 Endospore-forming: One of the defining characteristics of Bacillus
licheniformis is its ability to form heat-resistant endospores. These spores are
highly durable and can survive extreme conditions, including heat, desiccation,
and radiation. Spores are usually found in the terminal or subterminal position
within the cells.
1.2. Environmental factors
Bacillus licheniformis thrives in various environments and exhibits resilience under
harsh conditions due to its ability to form spores. Several environmental factors
influence its growth, survival, and activity. Below are the key environmental factors
affecting Bacillus licheniformis:
 Temperature:
Optimal growth temperature: Bacillus licheniformis is considered a moderately
thermophilic bacterium, with an optimal growth temperature of around 30°C to 50°C.
It can survive higher temperatures due to its spore-forming ability.
Heat resistance: The spores of Bacillus licheniformis can withstand temperatures
much higher than the growth range, enabling the bacterium to survive food
processing, such as baking, pasteurization, or cooking.
 pH:
pH tolerance: Bacillus licheniformis can grow across a wide pH range, with an
optimal pH of around 6.0 to 9.0. This tolerance allows it to thrive in various
environments, including neutral to slightly alkaline soils and food products.
It is less tolerant to highly acidic environments, which limits its growth in products
with a low pH (acidic foods).
 Oxygen Availability:
Facultative anaerobe: Bacillus licheniformis can grow in both the presence and
absence of oxygen. It prefers oxygen-rich environments (aerobic conditions), but it
can also grow in low-oxygen environments (anaerobic conditions), which helps it
survive in various ecological niches, including soil and food products.
 Water Activity (aw):
Bacillus licheniformis requires a moderate to high water activity level for optimal
growth. Like most bacteria, it thrives in environments where water activity is above
0.90. However, its spores are highly resistant to desiccation and can survive in dry
environments such as flour, grains, or dehydrated food products.
1.3. Types of Foodborne Illness
There have been rare cases where Bacillus licheniformis has been implicated in food
poisoning, although it is not a frequent cause. In these cases, the symptoms are similar
to those caused by other Bacillus species, including vomiting, diarrhea, and abdominal
pain. The severity of symptoms can vary based on the toxin load and the health status
of the person affected.
Similar to the diarrheal syndrome caused by Bacillus cereus, Bacillus licheniformis
may produce enterotoxins in food that lead to gastrointestinal symptoms. The
symptoms of this illness can include: Watery diarrhea, Abdominal cramps, Nausea
(sometimes), Occasional vomiting
These typically have an onset time of 2-14 hours and last no longer than 36 hours.
1.4. Control in Food
To extend the shelf life of bakery products, lowering water activity (aw) is essential,
which can be achieved by dehydration or adding sugars and salts to reduce available
water for microbial growth. Lower aw (around 0.75) significantly prevents mold but
must be balanced to avoid impacting product texture. Freezing at temperatures like -
18°C to -22°C can also preserve bakery items, maintaining freshness for months.
Additionally, using preservatives, such as sorbic acid or natural alternatives like
vinegar, helps control mold and microbial spoilage in baked goods.

References:
- De Candia, S., De Angelis, M., & Gobbetti, M. (2007). "Bacterial spoilage of
bakery products." - Trends in Food Science & Technology, 18(1), 13-20.
- Madigan, M. T., Martinko, J. M., & Parker, J. (2006). Brock Biology of
Microorganisms. 11th Edition. Pearson Prentice Hall.
- Magnusson, J., & Ström, K. (2003). "Growth inhibition of food-borne pathogens in
the presence of Bacillus licheniformis" - Journal of Food Safety, 23(3), 217-229.
- Granum, P. E. (2007). "Bacillus: Food poisoning." In M.P. Doyle & L.R. Beuchat
(Eds.), Food Microbiology: Fundamentals and Frontiers (3rd ed.). ASM Press.
- P.Saranraj and M.Geetha. (2012). “Microbial Spoilage of Bakery Products and Its
Control by Preservatives”. International Journal of Pharmaceutical & Biological
Archives

Reformulation involves a reduction of available water e.g., aw in bakery products to

obtain a longer shelf life. Reduction in product aw can be achieved by dehydration,
either through evaporation or freeze-dryingorbyhighosmoticallyactiveadditives e.g.,
sugars and salts, incorporated directly into the food. The degree of aw reduction is of
practical significance in making a food non-perishable. The response to a given degree
of aw varies greatly among microorganisms in different environments. Water
contained in solutions of sugars and salt becomes unavailable to microbes due to the
increased concentration of crystalloid. Furthermore, microbes are directly damaged
osmotically by concentrations of these substances. This effect may be due to the
adverse influence of lowered water availability on all metabolic activities, since all
chemical reaction of cells require an aqueous environment. Control of mold growth in
bakery products normally relies on maintaining a sufficiently low aw. For example, an
aw of 0.75 can give a 6 month extension in mold free shelf life. Higher aw levels e.g.,
above 0.77
will only result in a short extension of shelf life. However, since low aw can adversely
affect the quality of the product and cause changes in shape and texture, care must be
taken when reducing product aw .
5.2. Freezing
Freezing has been used for long term preservation of bakery products particularly,
cream filled products. Quick freezing is important in controlling the formation of ice
crystals. Large ice crystals are formed when the rate of freezing is slower; the large
crystals can disrupt membranes and internal cellular structures . Cakes, cookies,
short cake, and pancakes are commonly frozen and marketed in the frozen form.
Bread has been held fresh for many months by storage at -22°C. In contrast to fresh
bread, which stales in less than a week, frozen bread stales very slowly. Therefore, the
lower the temperature, the more slowly it stales. Desrosier (2006) reported that bread
frozen quickly after baking and held for one year at -18°C, was equivalent in softness
to fresh bread held for two days at 20°C.
5.3. Preservatives
Preservatives are most commonly used to control mold growth in baked goods. The
Code of Federal Regulations (CFR) defines preservatives “as an antimicrobial agent
used to preserve food by preventing growth of microorganisms and subsequent
spoilage”. There are two classifications of preservatives: chemical and natural
permitted chemical mold inhibitors in bread include acetic, sorbic, propionic acids and
their salts. Natural food preservatives, such as cultured products, raisins, vinegar, are
identified by their common name on the ingredient statement.