Microbiology of
fermented food
Dairy and other products
Fermentation has been a major way of preserving food over the last several
thousand years.
Microbial growth, either of natural or inoculated populations causes
chemical and or textural changes to form a product that can be stored for
extended periods.
The fermentation process is also used to create new, pleasing food flavors
and odors.
The major fermentations used in food microbiology are the lactic, propionic
and ethanolic fermentations.
These fermentations are carried out with a wide range of cultures, many of
which have not been characterized. E.g. in traditional settings
Fermented milks
Throughout the world, at least 400 different fermented milks are
produced.
These fermentations are carried out by mesophilic, thermophilic and
therapeutic lactic acid bacteria as well as by yeasts and molds.
Major examples of these fermentation types are discussed here.
Mesophilic milk fermentations
These result from similar manufacturing techniques, in which acid
produced through microbial activity causes protein denaturation.
To carry out the process, one usually inoculates milk with the desired
starter.
It is then incubated at optimum temperature (between 20 and 30oC),
and then stops microbial growth by cooling.
Starter cultures, bacteriophage
infections and plasmids
Cultures of lactic acid bacteria, called starter cultures, are added to milk during the
preparation of many dairy products.
For example, Streptococcus lactic and S. cremoris are used in the production of
cheese.
One of the greatest problems for the dairy industry is the presence of bacteriophages
that destroy these starter cultures.
Lactic acid production by a heavily phage-infected starter culture can come to a halt
within 30 minutes.
The industry has tried to overcome this problem by practicing aseptic techniques in order
to reduce phage contamination and by selecting for phage-resistant bacterial cultures.
Most efforts at control have not been successful in the long term.
It has been found that the very aseptic techniques and phage resistant pure cultures used
to attempt to solve this problem actually were part of the problem.
The most stable and dependable cultures called P starter cultures, contain the
bacteriophages in a pseudolysogenic state.
When cultures are grown without phages or under aseptic conditions (L-starters), they
lose their phage resistance.
It has been found that what is needed are certain plasmids that can block phage
adsorption.
The loss of the plasmids in a subpopulation of the bacteria allows the phage carrier state
to be established.
New phages can develop by acquiring restriction enzymes from plasmids.
These modified phages again become lytic, establishing a new equilibrium in the
population.
Other control approaches are being tested. Antisense RNA is now being used in an
attempt to provide an agent against bacteriophage genes to help in the constant struggle
between lactic acid bacteria and their phage.
The organisms Lactobacillus spp. and Lactobacillus lactis cultures are used
for aroma and acid production.
The organism Lactococcus lactis subsp. diacetilactic converts milk citrate to
diacetyl, which gives a special buttery flavor to the finished product.
The use of these microorganisms with skim milk produces cultured butter-
milk and when cream is used, sour cream is the result.
Thermophilic
In addition to mesophilic milk fermentations, thermophilic fermentations
can be carried out at temperatures around 45oC.
An important example is yoghurt production.
Yoghurt is made using a special starter culture in which two major bacteria
are present in a 1:1 ratio. Streptococcus thermophiles and Lactobacillus
bulgaricus.
With these two organisms growing in concert, acid is produced by
Streptococcus and aroma components are formed by Lactobacillus. Freshly
made yoghurt contains 109 bacterium per gram.
Therapeutic
Fermented milks may have beneficial therapeutic effects.
Acidophilus milk is produced by using Lactobacillus acidophilus.
L. acidophilus may modify the microbial flora in the lower intestine, thus
improving general health and it is often used as a dietary adjunt.
Many microbes in the fermented dairy products stabilize the bowel
microflora, and in some appear to have antimicrobial properties.
Exact nature and extent of health benefits are still unclear.
Major categories and examples of fermented milk products
Category Typical examples
1. Lactic fermentations
Mesophilic Buttermilk
Cultured buttermilk
Ymer
Langofil etc
Thermophilic Yogurt, laban, zabadi, Bulgarian
buttermilk skyr etc
Therapeutic Biogarde, Bifighurt, yakult etc
2. Yeast-Lactic fermentation Kefir, koumiss, acidophilus-
yeast milk
3. Mold-lactic fermentations Viili
However, they are thought to involve minimizing lactose intolerance,
lowering serum cholesterol, and possibly exhibiting anticancer activity.
Several lactobacilli have antitumor compounds in their cell walls.
Such findings suggest that diets including lactic acid bacteria, especially L.
acidophilus, may contribute to the control of colon cancer.
Another group used in milk fermentation are the Bifidobacteria. These are
irregular, non-sporing, gram-positive rods that may be club-shaped or forked
at the end.
Bifidobacteria are non-motile, anaerobic and ferment lactose and other
sugars to acetic and lactic acids.
They are typical residents of the human intestinal tract and were discovered
in the 1906.
Many beneficial properties are attribute to them.
They are thought to help maintain the normal intestinal balance, while
improving lactose tolerance; to possess antitumorigenic activity and to
reduce serum cholesterol levels.
In addition, some believe that they promote calcium absorption and the
synthesis of B-complex vitamins.
It has also been suggested that bifidobacteria reduce or prevent the
excretion of rotaviruses, a cause of diarrhea among children. Products
amended with this organism ae now available in many parts of the world.
Cheese production
Cheese is one of the oldest human foods and is thought to have been
developed about 8000 years ago.
About 2000 distinct varieties of cheese are produced throughout the
world, representing approximately 20 general types.
Often, cheeses are classified based on texture or hardness as soft cheeses
(e.g. cottage, cream etc), semisoft (e.g. Muenster, Limburger, blue) or hard
(e.g. cheddar, Swiss) or very hard cheeses (e.g. Parmesan).
All cheese results from a lactic acid fermentation of milk, which results in
coagulation of milk proteins and formation of a curd.
Rennin, an enzyme from calf stomachs, but now produced by genetically
engineered microorganisms can also be used to promote curd formation.
After the curd is formed, it is heated and pressed to remove the watery part
of the milk called whey, then it is salted and then usually ripened.
The cheese curd cam be packaged for ripening with or without additional
microorganisms.
Cheese curd inoculation is used in the manufacture of Roquefort and blue
cheese. In this case Penicillium roquefortii spores are added to the curds just
before the final cheese processing.
Sometimes the surface of an already formed cheese is inoculated at the start
of the ripening, for example Camembert cheese is inoculated with spores of
Penicillium camemberti.
The final hardness of the cheese is partially a function of the length of
ripening.
Soft cheeses are ripened for only about 1 to 5 months, whereas hard cheeses
need 3 to 12 months and very hard cheeses like Parmesan require 12 to 16
months ripening.
The ripening process is also critical for Swiss cheese. Gas production by
Propionibacterium contributes to the final flavor development and hole or
eye formation in this cheese. Some cheeses are soaked in brine to stimulate
the development of specific fungi and bacteria e.g. Limburger cheese.
Major categories and examples of fermented milk products
Category Typical Examples
Lactic fermentation Buttermilk
Mesophilic Cultured buttermilk
Langofil
Ymer
Thermophilic Yoghurt
Zabadi
Bulgarian buttermilk
Skyr
Therapeutic Biograde
Yakult
Acidophilus milk
Other fermented foods
Many other plant products can be fermented as summarized (see table).
These include sufu, which is produced by the fermentation of tofu, a
chemically coagulated soybean milk product.
To carry out the fermentation, the tofu curd is cut into small chunks and
dipped into a solution of salt and citric acid.
After the cubes are heated to pasteurize their surfaces, Actinimucor
elegans and other Mucor species are added.
When a white mycelium develops, the cubes, now called pehtze are
This product is a delicacy in many parts of the world, especially in the west.
Another product is tempeh, a soybean mash, fermented by Rhizopus.
Sauerkraunt or sour cabbage is produced from fermented cabbage. A
concentration of 2.2 to 2.8% sodium chloride restricts the growth of gram
negative bacteria while favoring the development of lactic acid bacteria.
The primary microorganisms contributing to this product are Leuconostoc
mesenteroides and Lactobacillus plantarum. A microbial succession occurs
in sauerkraut’s development.
The activities of the lactic acid producing cocci usually cease when the acid
content reaches 0.7 to 1%. At this point, Lactobacillus plantarum and
Lactobacillus brevis continue to function.
The final acidity is generally 1.6 to 1.8 with lactic acid comprising 1.0 to 1.3%
of the total acid in a satisfactory product.
Pickles are produced by placing cucumbers and such components as dill
seeds in casks filled wit brine.
The sodium chloride concentration begins at 5% and rises to about 16% in 6
to 9 weeks.
The salt not only inhibits the growth of undesirable bacteria but also extracts
water and water-soluble constituents from the cucumbers.
These soluble carbohydrates are converted to lactic acid.
The fermentation which can require 10 to 12 days involves the development
Leuconostoc mesenteroides, Enterococcus faecalis, Pedicoccus cerevisiae,
Lactobacillus brevis and L. plantarum. L. plantarum plays the dominant role
in this fermentation process.
Sometimes, to achieve more uniform pickle quality, natural microorganisms
are first destroyed and the cucumbers are fermented using pure cultures of
P. cerevisiae and L. plantarum.
Grass, chopped corn and other fresh animal feeds if stored under moist
anaerobic conditions undergo a lactic acid mixed fermentation that
produces a pleasant smelling silage.
Trenches or more traditional vertical steel or concrete silos are used to
store the silage. If anaerobic conditions are not maintained, the silage can
undergo rapid deterioration.
The probiotic concept
The large intestines of humans and animals contain a very complex and
balanced microbiota.
These microorganisms normally prevent infection and have a positive effect
on nutrition.
Any abrupt change in diet, stress or antibiotic therapy can upset this
microbial balance, making the host susceptible to disease and decreasing
the efficiency of food use.
Probiotics (Greek; pro, for and bios, life), the oral is the administration of
either living microorganisms or substances to promote health and growth.
Probiotics have the potential to reestablish the natural balance and return the
host to normal health and nutrition.
Probiotic microorganisms are host specific, thus a strain selected as a
probiotic in one animal may not be suitable in another species (hence the
feaces story).
Additionally, the microorganism selected for probiotic use should exhibit the
following characteristics:
1. Adhere to the intestinal mucosa of the host
2. Be easily cultured
3. Be non-toxic and non-pathogenic to the host
4. Exert a beneficial effect on the host
5. Produce useful enzymes or physiological end products that the host can
use.
6. Remain viable for a long time.
7. Withstand HCl in the host’s stomach and the bile salts in the small
intestines
There are several possible explanations of how probiotics probiotic
microorganisms displace pathogens and enhance the development of the
microbial balance in the large intestines. These include:
1. Competition with pathogens for nutrients and adhesion sites.
Inactivation of pathogenic bacterial toxins or metabolites.
Production of substances that inhibit pathogen growth.
Stimulation of non-specific immunity.
A wide variety of probiotic preparations have been patented for cattle, goats,
horses, pigs, poultry, sheep and other domestic animals.
Most of these preparations contain lactobacilli and/or streptococci; a few
contain bifidobacteria.
Evidence has accumulated that certain probiotic also offer considerable
health benefits for humans. Potential benefits include:
1. Anticancer activity
2. Control intestinal pathogens e.g. Clostridium dificile.
3. Improvement of lactose use in individuals who have lactose intolerance.
4. Reduction of serum cholesterol concentration.
Although the development of probiotics is still in the early stages, a better
understanding of the normal microbiota in the large intestines of both
animal and humans will be forthcoming as more microbiologists investigate
probiotic activity.