Epidemiological Studies on Fermented foodsby:Tuhin Kumar DasguptaIMTH-8th SemesterKSBT

History of Fermentation

• Earliest evidence of production of fermented beverages dates from 8000 years ago in the Caucasus region.

• 5000 years ago, Chinese building ‘The Great Wall’ were eating fermented mixed vegetables.

• Buddhist’s excluded Meat from their diet, which played a vital role in the budding the science of fermentation.

• Development of sophisticated soybean products was a result of Buddhist monks working in their monasteries.

• Asian cultures pioneered ways to produce meat-like flavors and textures from vegetable proteins.

• Much of economically developing world depends on various fermented foods as staples of their diets

• Louis Pasteur (1822–1895), during the 1850s and 1860s, showed that fermentation is initiated by living organisms in a series of investigations.

• In 1857, Pasteur showed that lactic acid fermentation is caused by living organisms.

• In 1877, working to improve the French brewing industry, Pasteur published his famous paper on fermentation, "Etudes sur la Bière", which was translated into English in 1879 as "Studies on fermentation"

FERMENTED FOODS…an overview

Name Origin Description

Idli India Staple food of South India; prepared by steaming a fermented batter of rice and black gram (Phaseolus mungo); Leuconostoc mesenteroides present in blackgram

Bagoong Philippines Philippine condiment made of partially or completely fermented fish (bagoóng isdâ) or krill (bagoóng alamáng) and salt; fermentation process also results in fish sauce

Kefir Caucasus Mountains, Turkey

Traditionally, Kefir was made by mixing Kefir grains with fresh, raw, cow or goat milk which was then left in goatskin leather bags to ferment. Saccharomyces kefir, Lactobacillus caucasicus used

Kimchi Korea made by fermenting vegetables with Lactic acid bacteria; often allowed to ferment underground in jars for months; varieties of kimchi made from napa cabbage, radish, scallion, or cucumber

Kombucha Asia fermenting tea with principal yeast Schizosaccharomyces pombe; Principal bacteria are Acetobacteria xylinum, Gluconobacter bluconicum

Miso Japan fermenting soybeans with salt and Aspergillus oryzae, and sometimes rice, barley; thick paste used for sauces and spreads

Name Origin Description

Nattō Japan Japanese food made from soybeans fermented with Bacillus subtilis.

Sauerkraut German Finely cut cabbage that has been fermented by various lactic acid bacteria, including Leuconostoc, Lactobacillus, and Pediococcus

Şalgam Turkey popular beverage made with the juice of red carrot pickles, salted, spiced, and flavoured with aromatic turnip fermented in barrels with the addition of ground bulgur.

Douchi China fermented and salted black soybean; Unlike some other fermented soybean-based foods such as natto or tempeh, douchi is used only as a seasoning, and is not meant to be consumed in large quantities, being typically much saltier

Tofu/ stinky Tofu

Taiwan fermented tofu that has a strong odor; it does not have a fixed formula for starter bacteria; The traditional method for producing stinky tofu is to prepare a brine made from fermented milk, vegetables, and meat

Tempeh Indonesia A traditional soy product originally from Indonesia. It is made by a natural culturing and controlled fermentation process that binds soybeans into a cake form

Name Origin Description

Pro-biotic Yogurt/ Dahi

Turkey Milk fermented by live cultures so as to enhance its nutritional value and digestibility. The best yogurt is low fat with a creamy, slightly sweet texture which has been fermented with one part L. bulgaricus to seven parts S. thermophilus, these being the best yogurt cultures

Cheese Cheese is also a milk fermentation, but its production is more complex. Different bacteria come into play and production periods are much longer than yogurt. Despite there being 20 classes and hundreds of varieties of cheeses the initial manufacturing process is surprisingly similar.

There is no conclusive evidence indicating where cheese-making originated, either in Europe, Central Asia, the Middle East, or Sahara.

PRODUCTION PROCESS

MISO – bean paste• Japanese name to paste-like products from fermentation

of cereal, soybeans and salt with molds, yeasts and bacteria

• ‘chiang’ in China, ‘doenjang’ in Korea• 590,000 MT of commercial miso & 150000 MT of

homemade miso are produced in Japan annually• 3 types of Miso based on Raw materials used: 1) Rice Miso

(rice, soybean & salt) 2) Barley Miso (barley, soybean & salt 3) Soybean Miso (soybean & salt)

• 80% of total Industrial production is Rice Miso

Polished rice

Washed

Soaked in water

Excess water drained

Steamed

Cooled

Inoculated with Aspergillus oryzae & incubated

Koji (Mold rice)

Soybeans

Washed

Soaked in water 20hr/16°C

Excess water drained

Steamed 115°C/20min

Cooled

Mixed

Fermented

Transferred

Ripened

Blended & Mashed

Pasteurized Packed Miso

Saccharomyces rouxii, Pediococcus halophilus

NaCl

TYPES OF MISO IN JAPAN

Miso has a strong buffer activity due to the presence of proteins, peptides, amino acids produced during fermentation. This property plays an important role as a seasoning in variety of foods.

3300 kg light yellow salty miso with a moisture content of 48% is made from 1000 kg of soybeans, 600 kg of rice and 430 kg of salt

TEMPEH• ‘tempe kedelee’ as called in Indonesia, made by fermenting

dehulled soybeans with a mold, Rhizopus oryzae or Rhizopus oligosporus; the mycelia bind the soybean cotyledons in a cake-like texture

• Rhizopus oligosporus is highly proteolytic• Highly acceptable to people of the Western world• Used as main dish or meat substitute in Indonesia• High protein content• Small pieces of tempeh from previous culture are mixed with

soybeans, which are then wrapped in banana leaves• Fermented at room temperature for 24-48 hours until the beans

are covered with white mycelium

Dehulled full-fat soybean grits

Soaked in Tap water, 30min/25°C

Drained

Cooked (30min)

Drained & cooled

Inoculated spore suspension of Rhizopus oligosporus Saito NRRL 2710

Packed in petri dishes

Incubated @ 31°C for 20-24hrs Tempeh Cake

Tempeh- after fermentation

• soluble nitrogen increases from 0.5%to 2%• Free amino acids in tempeh increases• Niacin (Vit B3), riboflavin (Vit B2), pantothenic acid (Vit B5) and

Vitamin B6 contents of soybean increases• Thiamin does not change significantly• Lipids in tempeh were found to be more resistant to auto-oxidation

than in soybeans• In 1964, György et al. isolated a new isoflavone from tempeh

designated as “Factor 2”, which was identified as 6,7,4-trihydroxyisoflavone.

• György also found antibacterial agents in tempeh, which were active against Gram +ve bacteria such as Bacillus subtilis, Staphylococcus aureus

• Stachyose in soybeans was found to decrease as fermentation progressed

What’s New…

NATTO• Natto fermentation predominated by Bacillus natto (Gram

+ve, rod shaped, aerobic), identified as Bacillus subtilis• Persistent musty flavor of the bacteria, covered with

viscous, sticky polymers that Bacillus subtilis produces• Japan produced 12,390,000 tonnes of natto last year (up

35.9% over 2013)• Quality is ascertained by the stickiness of the beans• Rice straw is credited for supplying the fermenting

organism & also providing the aroma of the straw, with absorbing ammonia odor from natto

• 60% moisture content, has short shelf life• 2 types of Bacillus natto in laboratory of Food Research

Institute, Ministry of Agriculture and Forestry, Tokyo, Japan-

1) one with optimum temperature from 30° - 45°C 2) other, from 35° - 45°C

YOGURT• ‘Yogurt’ comes from Turkey and refers to a tart, a product

of fermented milk. • Lactose in milk is fermented to lactic acid• Greek yogurt starter is an example of a heat loving

(thermophilic) culture. Filmjolk is an example of a cooler temperature (mesophilic) culture.

• In 1981, the FDA defined fresh, prepared yogurt in the US and stated that it must include Lactobacillus bulgaricus and Streptococcus thermophilus

Historical Facts:• Recorded history states that Genghis Khan, the founder of the Mongol Empire, and his armies lived on yogurt.• The first industrialized production of yogurt is attributed to Isaac Carasso in 1919 in Barcelona – his company “Danone” was named for his son, “Little Daniel”.

Production of yogurt starts by conditioning the milk:

Water content of milk is first lowered 25% by evaporation and 5% milk solids are added.

As a final conditioning step, the milk is heated to 86 to 93°C for 30-60 minutes. (This causes some breakdown of proteins and other molecules and kills contaminating microbes that may compete with the starter culture.) After cooling to 45°C, a 1:1 mixture of Streptococcus thermophilus and Lactobacillus bulgaricus is added.

Fermentation is at 45°C until the desired degree of acidity is reached. This usually occurs in 3-5 hours.

The finished product may have other ingredients added (such as mold inhibitors or dye) and is packages with fruit.

Yogurt is stored at 0-4°C until consumed to prevent spoilage.

Yogurt with fruit on the bottom was first introduced in 1947 by Dannon.

Proposed Health benefits stemming from probiotic consumption

The ‘Gut’ feeling

SAUERKRAUT (fermented cabbage)

• Captain Cook used cabbage (Brassica oleracea) fermented in lime juice to prevent scurvy, and the Romans consumed sauerkraut to prevent stomach infections in Ancient times

• results from Lactic acid fermentations of shredded cabbage by Leuconostoc mesenteroides, Lactobacillis brevis & Lactobacillus plantarum

• An activating ingredient is added (salt or vinegar) which draws the natural moisture from the food to form a brine.

• Concentration of salt must not be less than 2%, nor more than 3%• Salt extracts water from the shredded cabbage by osmosis, forming

fermentation brine• Salt suppresses the growth of undesirable bacteria• Too much salt delays fermentation• Neither refrigeration nor pasteurization is required

Phases of Fermentations:

• First phase, anaerobic bacteria such as Klebsiella and Enterobacter lead the fermentation, and begin producing an acidic environment that favours later bacteria.

• Second phase starts as the acid levels become too high for many bacteria, and Leuconostoc mesenteroides and other Leuconostoc spp. take dominance.

• Third phase, various Lactobacillus species, including L. brevis and L. plantarum, ferment any remaining sugars, further lowering the pH.

• Properly cured sauerkraut is sufficiently acidic to prevent a favorable environment for the growth of Clostridium botulinum, the toxins of which cause botulism

KOMBHUCHA

• Kombucha mushroom, a fermented yeast enzyme tea, originated in Asia during the Chinese Tsin dynasty in 212BC.

• Known as the tea of immortality. Today the tea - once routinely used by Samurai - is widely used again in Japan.

• a probiotic, microbiological food. It helps regenerate the bowel flora and is excellent for wellbeing

• principal yeast is Schizosaccharomyces pombe, & principal bacteria are Acetobacteria xylinum, Gluconobacter bluconicum

• Kombucha culture feeds on the sugar and, in exchange, produces other valuable substances which change into the drink: glucuronic acid, glucon acid, lactic acid, vitamins, amino acids, antibiotic substances, 0.5% to 1% alcohol is produced as well as other products

• commonly referred to as ‘SCOBY’ drink

What does ‘SCOBY’-DO…

Sometimes inaccurately called mushroom tea or fungus tea, it’s based on a fermenting agent commonly referred to as SCOBY: “symbiotic culture of bacteria and yeast.”

A slab of this admittedly kind of ugly looking culture is placed in a glass of sweetened black tea to create kombucha. The finished drink is fizzy, acidic and slightly alcoholic.

Me too…

Study reveal’s… The American Cancer Society (ACS) points out that kombucha tea,

like many fermented foods, “has been promoted as a cure-all for a wide range of conditions including:

• baldness, insomnia, intestinal disorders, arthritis, chronic fatigue syndrome, multiple sclerosis and cancer.

• Supporters say that Kombucha tea can boost the immune system and reverse the aging process.

• Kombucha tea is said to contain antioxidants, compounds that block the action of free radicals (activated oxygen molecules that can damage cells).

• For people who have cancer, proponents claim the tea can improve the body’s defenses (especially in the early stages of cancer) by detoxifying the body and enhancing the immune system. After the body has been detoxified, the tea is said to help repair and balance the body and fight off disease.”

A Healthy Boon… Fermented

foodHealth benefit

Tempeh

i. Fermentation increases the digestibility of soy (especially its proteins), nutrient absorption from soy (including absorption of phytonutrient isoflavones like genistein and daidzein), and the concentration of bioactive peptides (formed during the breakdown of soy proteins during fermentation).

ii. Replacing meat and dairy with tempeh and other soy products would also lower our total cholesterol intake by about 125 milligrams per day and our saturated fat by about 2.4 grams per day.

iii. These nutritional changes, in turn, would lower our risk of several chronic diseases, including cardiovascular diseases.

iv. Proteins and Peptides: defensins , glycinin , conglycinin , lunacin v. Phenolic Acids: Caffeic acid, Coumaric acid, Ferulic acid, Gallic acid vi. The most consistent effect of soybean intake on blood fats has been a

moderate lowering of LDL cholesterol.vii. soyasaponins can lessen the rate of lipid peroxidation in blood

vessels

Fermented food

Health benefit

Sauerkraut

i. a source of vitamins C, B, and Kii. fermentation process increases the bioavailability of nutrients

rendering sauerkraut even more nutritious than the original cabbage.iii. it is also low in calories and high in calcium and magnesiumiv. good source of dietary fiber, folate, iron, potassium, copper and

manganesev. If unpasteurized and uncooked, sauerkraut also contains live

lactobacilli and beneficial microbes and is rich in enzymes. The fiber and supply of probiotics improve digestion and promote the growth of healthy bowel flora, protecting against many diseases of the digestive tract.

vi. Sauerkraut has been used in Europe for centuries to treat stomach ulcers

vii. The October 23, 2002 issue of the Journal of Agriculture and Food Chemistry reported that Finnish researchers found the isothiocyanates produced in sauerkraut fermentation inhibit the growth of cancer cells in test tube and animal studies.

viii. A Polish study in 2010 concluded that "... Induction of the key detoxifying enzymes by cabbage juices, particularly sauerkraut, may be responsible for their chemopreventive activity demonstrated by epidemiological studies and in animal models

Fermented food

Health benefit

Kombucha

i. The intestinal microflora is carefully balanced. The human gut is home to around 400 different species of good and bad bugs. However, the good bugs have to share their environment with bad bugs such as Salmonella, E.Coli and Clostridium. The way to balance between good and bad bacteria is to maintain good digestive health, creating stable "microflora". It is generally recommended that probiotics are taken on a daily basis to create this balance.

ii. Kombucha being one of the best skin treatments for acne and an arthritis health drink.

iii. Organic Kombucha Tea is considered by some as a delicious 'Cancer Foods‘

Natto

Natto, is a natural energy food that is: i. High in Protein ii. Good Source of Dietary Fiber iii. Rich in Vitamin K2 & B2 iv. Contains Calcium & Iron v. The unique enzyme in Natto, Nattokinase, may be potentially useful as a

clot-buster and blood thinnervi. A 2009 study showed that nattokinase may be effective in catabolism of

toxic amyloid fibrils associated with Alzheimer's Disease, as well as the insulin fibrils associated with diabetes

Researchers from Murdoch Children's Research Institute found that in 80 % of test cases, children were able to eat small amounts of peanuts with no adverse effects if they also ate probiotics. Thus far, probiotic treatment against peanut allergies had only worked in lab mice.

Journal of Physiological Anthropology reveals…

• gut-to-brain communication• fermented foods could lead to improved neurotransmitter and

neuropeptide production in the brain• animals had alterations in the system of the brain concerning the

neurotransmitter gamma-Aminobutyric acid (GABA) that were similar to the effects of antidepressants

• supplementation with Bifidobacteria, also found in yogurt appears to attenuate an exaggerated stress response and maintain adequate levels of the neuropeptide brain-derived neurotrophic factor (BDNF)

• Bifidobacteria supplementation also appears to decrease monoamine oxidase activity in the brain, potentially increasing neurotransmitter levels between synapses

• The supplement also lowered levels of the stress hormone cortisol compared with placebo. When given to rodents, the same supplement reduced “behaviors indicative of anxiety.”

Thank You!!!