Human's probiotic - Lactic acid bacteria

According to Dr. Ken Rifkin, “The average human body contains approximately three and a half pounds of bacteria some of which perform essential functions AND others which promote disease.” When the equilibrium of ‘friendly’ and pathogenic bacteria is disturbed through the ingestion of chemical additives, birth control pills, antibiotics, alcohol, pesticides, food additives and even stress — disease producing bacteria will multiply within the intestinal tract.”

Various pharmaceutical companies sell different strains of Lactobaccilus acidophilus and B. bifidiumin order to help replace our valuable microflora.

The interest of man for probiotics is not new; one hundred years ago Metchnikoff suggested in his book entitled “The prolongation of life” that consuming lactobacilli  that survive in the intestinal tract was desirable for health. Bulgarians were then known for their longevity which Metchnikoff attributed to their consumption of lactobacilli  from fermented milk product.

 

Diffrent morphological pattern of Lactobacillus Sp

 

 

 

Morphogical pattern of bfidobacteria sp.

 

Following the discovery of antibiotics after World War ΙΙ, the popularity of probiotics decreased, but they were still used to reestablish the intestinal microflora following aggressive antibiotic treatments. It is only recently that there is a renewed interest in the use of probiotics for humans and animals and for the understanding of their mode of action. The renewed interest in probiotics has emerged from a general public and scientific concern about the widespread use of antibiotics and the possibility for transfer of antibiotic resistance to human pathogenic bacteria. For this reason the European Union banned the use of antibiotics for non-therapeutic purposes in January 2006. In Canada, there is still no legislation to this effect, but it is probably a question of time. It is therefore imperative to find safe alternatives to the use of antibiotics.
                
Various types of probiotic product that contain lactobacillus sp. Which is better for you?:)
From time to time a pharmaceutical company will do a definitive paper on some subject. Such contributors do not choose to be credited, however, because of possible trouble with the FDA -- accused of making claims that their product will do such and such in the absence of a $25,000,000 double-blind study to “prove” “safety” and “effectiveness” -- or because folks may not take their information seriously because they sell the product and have the appearance of a conflict of interest. Sometimes, it is true, a company shows only their good points and deflates bad parts, just to sell their own product, and so pharmaceutical promotional literature is suspect.


Intestinal Flora Competition
Intestinal Flora Competition among microorganisms in the large intestine is a major consideration since the highest numbers of bacteria occur here.

Bifidobacteria are the predominant organisms in the large intestine of breast-fed infants, accounting for about 99% of the cultivatable flora.

Lactobacilli, Enteterocci, and Coliforms comprise about 1% of the flora. “Bifidobacteria are a major component in the large intestine of adolescents and adults, while Lactobacilli, Enterococci, and Coliforms are a smaller component of the flora. Bifidobacteria are reduced significantly in the stools of old people, but Clostridia, Streptococci, and Coliforms are increased. “Lactobacilli are the predominant organisms in the small intestine. Lactobacilli have important metabolic activities, although they may occur in smaller numbers than Bifidobacteria in the upper and lower intestines combined.


The Role of Lactobaccilli and B. bifidium in the Intestinal Tract
 
1) Antibiotic Production
Inhibits the pathogenic flora by production of the following antibiotics
Lactolin (L. plantarum)
Lactobrevin (L. brevis)
Bulgarican (L. bulgaricus)
Acidophilin (L. acidophilus)
Lactocidin (L. acidophilus)
Acidolin (L. acidophilus)
Lactolin (L. acidophilus)2,12,13

2) Organic Acid Production
Lactobacilli produce lactic acid. Organic acetic and lactic acids which are produced by lactic acid bacteria will inhibit the growth of many bacteria, especially pathogenic gram-negative types. Lactobacillus acidophilus produces DL-lactic acid which is metabolized to a limited extent

3) Lower pH and Oxidation Reduction
Inhibition of pathogens by Lactobacilli is attributed to the lowering of the pH values by the liberation of acids, resulting in  antimicrobial action (altering oxidation-reduction potential) attributed to the lowering of the pH values by the liberation of acids, resulting in antimicrobial action (altering oxidation-reduction potential)

4) Competitive Antagonists
Lactobacilli may outcompete other bacteria for nutrients and occupy the sites, making them unavailable to other microorganisms. In particular, Lactobacilli consume certain B-vitamins and biotin, decreasing their availability for other organisms.

5) Bile Deconjugation
The role of Lactobacilli in the deconjugating of bile acids was studied. The results indicated that Lactobacilli can liberate (deconjugate) free bile acids in the intestinal tract and can exert an influence on the balance of bacteria present. [Deconjugation is the chemical process of separating the two amino acids taurine and glycine from bile acids, the bile then being recycled for reuse.

6) Detoxification
“The roles of B. bifidium and lactulose in the detoxification of subjects with chronic liver disease has been studied. The results showed that B. bifidium with lactulose may assist in re-establishing the normal intestinal flora which is usually disturbed in chronic liver cirrhosis. This is accomplished by a reduction of ammonia and free

Conclusion
“Certain microorganisms such as Lactobacillus acidophilus and Bifidobacterium species can help to maintain a favorable intestinal microflora which has been associated with good nutrition and health. “Bifidobacterium are the predominant organisms in the large intestine of breast-fed infants. They decrease into adulthood and diminish with old age. “Although Lactobacilli occur in smaller numbers than Bifidobacteria overall, they have important metabolic properties, especially in the small intestine where they predominate. “When taken in combination, a more complete, favorable intestinal microflora is achieved. The regular ingestion of Bifidobacteria with Lactobacilli will suppress harmful bacteria.

Lactobacillus acidophillus

Well, the name of acidophilus has no longer sounded strange for us; there are many types of fermented dairy products that use L. acidophilus. The most familiar to us are milk and yogurt product - Yakult and Vitagen; and the Korean fermentative vegetables – Kimch. It has been long suggested that L. acidophilus is a beneficial or so-called "friendly" bacteria for human, animal and plant health.

I trusted in Yakult, because it really showed the beneficial effect to me. There was a time it worked in my auntie intestinal disturbance, where my auntie having a bad acute diarrhea.

A bottle of Yakult 2 times a day really helps rapidly in recovering the intestinal digestibility function. Within 6 hours, my auntie’s diarrhea problem disappeared. Here, I just want to give a credit to Yakult and his founder - Dr. Shirota. And the truth is the lactobacillus strain in Yakult was named by his name – Lactobacillus Shirota.


My experiment

My experiment of culturing lactobacillus acidophillus serum

In this experiment, L. acidophilus was solely cultured in milk: culture of these bacteria alone in milk is simple and also the bacteria in commercial fermented milk and commercial probiotics may not be alive and/or the number of live bacteria may be lower than the products label

L. acidophilus was cultured in sterile milk. For sterilization, low fat milk was heated to 120°C for 20 minutes, cooled to 37°C and 2 percent of the L. acidophilus starter was added and incubated at 37 to 40°C for 18 hours. After incubation, the product was stored at 15°c until use. On the average, the fermented milk contained 2×108 cfu g-1 L. acidophilus.

I drink the fermented milk to treat my indigestion, it works well.:) Somehow, i used the residue to make plant soil inoculant! Effective!!


Morphologic and biochemistry (wikipedia)
L. acidophilus is a member of one of the eight main genera of lactic acid bacteria. Each genus and species have different characteristics but they are generally chained cocci or rod shaped gram positive , nonmotile, nonsporulating bacteria that produce lactic acid as a major or sole product of fermentative metabolism and use lactose as their main source of carbon to produce energy. L. acidophilus grows in or without the presence of oxygen, is able to live in highly acidic environments of pH 4-5 or lower and is characterised as a homofermentative ie produces lactic acid as its sole product.

Reproduction
Lactobacillus acidophilus reproduces by the process of binary fission. Binary fission is the form of asexual reproduction in single-celled organisms by which one cell divides into two cells of the same size, used by most prokaryotes. Asexual reproduction is the division of one cell into two cells.

Tempeh Fungus starter - Rhizopus oligosporus

What is tempeh?
Tempeh is a popular fermented food in Indonesia and Malaysia which is rich in nutrients and active substances. In my country, it’s one of the common foods that found in Malay’s restaurant and hawker center.

The first time I tasted it, I felt bit uncomfortable because of its odor smell. Actually, high quality tempeh has a mild “mushroomy” or “yeasty” aroma, can be sliced or cubed easily without crumbling. But, when I start to understand some significance of soybean fermentation in tempeh on the vitamins, amino acids, nutritional quality, functional types of microbes, and physico-chemical properties, I begin to love it.

The flavor of soy tempeh is full-bodied. It can be a “star” in any kitchen because it cooks quickly and can be prepared in hundreds of ways, using all cooking methods.

Recently, the consumption of Tempeh has been increasing rapidly, not only in Malaysia and Indonesia but also in the United States and Europe. Although Tempeh is not likely to be exported, the product is consumed and produced in many countries.


The making of tempeh
Tempeh is a fermented product made from soybeans that have been soaked and cooked to soften them. Like sour dough bread, tempeh requires a microbes inoculum starter, which is added to the cooked beans.
This mixture is left for 24 hours and the result is a firm textured product with a somewhat nutty flavor and a texture similar to a chew mushroom. Because Tempeh is firm and it can be formed into a patty, it is used as a substitute for animal products in what in the West is typically called as "mock" burgers or sandwiches.
This soya product is especially popular in Indonesia and is considered a national specialty. It has the necessary characteristics of a dietary staple in that is high in protein and fiber and is rich in other nutrients. It also has the advantage of containing Vitamin B-12, which is a by-product of the fermentation process . In this heat paper the research work carried out on tempeh fermentation and tempeh based products is reviewed.

Role of Microorganisms in Tempeh Production:
The type of microorganisms involved in tempeh production has been reported by many researcher. Tempeh is the result of mixed culture fermentation by a diverse group of microorganisms including moulds, yeasts, lactic acid bacteria and different gram-negative bacteria. Rhizopus oligosporus is the dominant tempeh fungus although some other moulds, such as R. oryzae and Mucor spp, may also contribute to the flavour, texture or nutritive value.

A researcher named Heseltine isolated many fungi from different lots of tempeh made in Indonesia and found that only Rhizophus could make tempeh in pure culture fermentation. They also found that the 40 strains of Rhizophus studied, 25 of them are R.oligosporus others are R.stolonifier，R.arrhizus.R.oryzae, R.formosaensis and.This finding was also confirmed by another researcher Saono , who isolated 118 cultures from 81 tempeh samples collected from markets in various parts of Indonesia. Mulyowidarso et al also reported variable growth of bacteria during fermentation of soybeans into tempeh with R.oligosporus.

There is also another study reported that Klebsiella pneumonia was responsible for the production of vitamin B12 in commercial tempeh. Yeasts are frequently detected in tempeh, but their role is still unknown
Rhizopus moulds produce natural, heat stable antibiotic agents against some disease-causing organisms. Indonesians who eat tempeh as a regular part of their diet recognize it as a medicine for dysentery and rarely fall victim to the intestinal diseases to which they are constantly exposed.



My experiment of making tempeh

Soak the soybeans water for 1whole night.

1) Put the beans in a cooking pot and add enough water to cover them.  Cook for 30 min. Drain off the water and dry the soybeans by continue heating them in the pot on medium heat for a few minutes and until the beans are dry. Allow the soybeans to cool down to below 35°C.

Inoculating the soybeans with tempeh starter

2) Sprinkle the soybeans with 1 teaspoon of tempeh starter. Mix with a clean spoon for about 1 minute to distribute the tempeh starter evenly. It's very important to mix the tempeh starter very well: it reduces the risk for spoilage and the fermentation will be faster.

 3) Perforate plastic cover with holes at a distance of about 1 cm by a thick but sharp needle. A normal needle is too thin, you need a fat needle or small nail (about 0.6 mm in diameter). This will allow the mould to breathe. Divide the soybeans in the two bags and seal them. Press them flat, making sure that the total thickness of the beans is max 3 cm.

 

incubating in room temperature (30'C) in my incubator

After 18 hours, the white formed

4)Place the packed beans in an incubator at 30°C or at a warm place for about 36- 48 hours during which the tempeh fermentation takes place.

Done

5) Then the container was filled completely with white mycelium and the entire contents can be lifted out as a whole piece.

Japan's National Microbes - Aspergillus oryzae

Aspergillus oryzae - the Japan's National Microbes!

Why? What make this species became the most important microbes of world-class quality assured country Japan?

Weeks ago, a friend from China told me they used A.oryzae to ferment porcine blood to serve as high-nutrient blood meal for his pigs ( due to the high proteolytic protease released from A.Oryzae that able to convert blood protein to highly digestible and nutrient value-added amino acids).
He said this method not only reduce feeding cost ( blood are reused instead of disposing), but also reduce environmental pollution! In China, this is a very common practice, but in Malaysia it seemed not that practical.

In Malaysia - we use Aspergillus oryzae to make soy souce!

In my living area, Soy sauce manufacture is carried out by traditional Chinese methods. Most of the factories are small-scale enterprises, producing less than 100 kiloliters per year.

Traditional soy sauces are made by mixing soybeans and grain with mold cultures such as Aspergillus Oryzae and other related microbes and yeasts (the resulting mixture is called "koji" in Japan; the term "koji" is used both for the mixture of soybeans, wheat, and mold; as well as for only the mold). Historically, the mixture was then fermented naturally in giant urns and under the sun, which was believed to contribute additional flavors. Today, the mixture is generally placed in a temperature and humidity controlled incubation chamber

Family member of Aspergillus
1) A. flavus and A. parasiticus are known to produce the potent carcinogen aflatoxin. ( this is the thing that making the livestock farmer terrified)

2)A. oryzae and A. soji have been used for producing food grade amylase and fermentation of oriental foods for centuries (Sooriyamoorthy et al., 2004; Geiser et al., 1998).
They are defined by the production of spore chains in radiating heads which range in color from yellow-green to olive brown. ( like the shape of nuclear explosion)

They are are highly aerobic and are found in almost all oxygen-rich environments, where they commonly grow as molds on the surface of a substrate, as a result of the high oxygen tension. Commonly, this fungi grow on carbon-rich substrates such as monosaccharides (such as glucose) and polysaccharides (such as amylose). Aspergillus species are common contaminants of starchy foods (such as bread and potatoes), and grow in or on many plants and trees. Members of the genus are also sources of natural products that can be used in the development of medications to treat human disease.

Use and application
1) Generally, It is used in Chinese and Japanese cuisine to ferment koji. Koji is a culture prepared by growing either Aspergillus oryzae mold on cooked grains and/or soybeans in a warm, humid place. Koji serves as a sourceof enzymes that break down (or hydrolyze / digest / split)natural plant constituents into simpler compounds when making miso, soy sauce, sake, amazake, and other fermented foods. Its fragrant white (or red) mycelium, which lookssomewhat like the surface of a tennis ball, has a delightfularoma resembling that of mushrooms. It is also used to saccharify rice, other grains, and potatoes in the making of alcoholic beverages such as huang jiu, sake, and shōchū.

2) An inexpensive and readily available agro industrial substrate such as rice bran can be used to produce cheap commercial enzymes like protease, lipase and amylase by solid-state fermentation.

3) Also, production of enzyme, antibiotic in various industry fields.

In conclusion, A. oryzae is long considered safe in food industry and recently being selected as eligible candidate of probiotics

Greatest threat for aquaculture's farmer

When you ask what is the greatest threat for aquaculture's farmer, perhaps if your fish know how to talk, i am sure the fish will say- Ammonia!!

What is ammonia? and why the rise and fall of  aquaculture industries are highly dependable on effective control of ammonia?

Before I go into detail of those Nitrifying and denitrifying bacteria, let me give a short brief introduction of nitrogen cycle and water nitrification. If u want to be successful in aquaculture farming, you must understanding  a very importance concept - nitrogen cycle.

Nitrogen cycle
Ammonification ------>Nitrification--------->  Denitrification.

1) Ammonification. While traveling through sewer pipes, the majority of the nitrogen contained in raw sewage (urea and fecal material) is converted from organic nitrogen to ammonia through a process called hydrolysis, accomplished by putrefying bacteria and fungus. Technically, in the majority of situations, more ammonium than ammonia is created during ammonification.The actual ratio is influenced by pH and temperature.

2) Nitrification. The biological conversion of ammonium to nitrate nitrogen is called Nitrification. Nitrification is a two-step process. Bacteria known as Nitrosomonas convert ammonia and ammonium to nitrite. {2NH3 +3 O2 → 2HNO2 +2 H2O +158 kcal (660kJ). }
Next, bacteria called Nitrobacter finish the conversion of nitrite to nitrate. The reactions are generally coupled and proceed rapidly to the nitrate form; therefore, nitrite levels at any given time are usually low.        {HNO2 + 1/2 O2 = HNO3 - ⊿ G = 18 kcal. }

http://nmlc.org/2011/06/the-critter-tank-and-the-nitrogen-cycle/
As we can see the above chemical equation, These bacteria known as “nitrifiers” are strict “aerobes,” meaning they must have free dissolved oxygen to perform their work. Nitrification occurs only under aerobic conditions at dissolved oxygen levels of 1.0 mg/L or more.
These two types of bacteria obtain the energy from the above oxidation process, but the energy utilization is not high, resulting in slow reproduction time. (i.e  more than 10 hours)

Today, people have not yet found a nitrification bacteria are able to directly convert the ammonia into nitrate!!

So to say, nitrification can only be completed by the combined effect of these two types of bacteria. We knew that the ammonia is harmful to human and fish, How about nitrite? Nitrite poisoning inhibits the uptake of oxygen by red blood cells - known as brown blood, eventually cause severe dealth in fresh water fish. Besides, nitrite can react with metal ions forming salt, and this salt tends to combine with amines, eventually forming a substance with strong carcinogenic effect- alkylene nitramine. Understanding the lethal effect of ammonia and nitrite residue, we have to manipulate the effectiveness of those beneficial bacteria, which in turn increases the efficiency of the nitrogen cycle.

Nitrosomanas

Nitrobacter

3.Denitrification. The biological reduction of nitrate (NO3) to nitrogen gas (N2) by facultative heterotrophic bacteria is called Denitrification. “Heterotrophic” bacteria need a carbon source as food to live. “Facultative” bacteria can get their oxygen by taking dissolved oxygen out of the water or by taking it off of nitrate molecules.


Role of nitrifying bacteria in Aquaculture?
Nitrifying bacteria highly demand oxygen to survive, they love to stay in a variety of fiber cotton, glass rings (silicone made), ceramic microporous filter, where they will get a higher concentration of oxygen exchange. Here is a very important key note for those want to begin their aquaculture farming. The presence of toxic substances in the breeding pool, mainly ammonia and nitrous acid (nitrite), but thank God that this two toxic substances consumed by nitrifying bacteria, and generates a non-toxic nitrate, nitric acid where these substances can directly absorbed by water algae and plankton.
From the ecological point of view of the water, we cannot prevent the ammonia generated.

The more the loading of organic food, the more ammonia generated!!

But you can try to increase the number of nitrifying bacteria to consume the increasing ammonia in the water. Many people disregard the importance the issue, thinking that the more input of the food, the more fast growing of the fish, eventually resulting in losing tones of money.

So, what should we do to improve the number of nitrifying bacteria?
The first thing is we must get nitrifying bacteria a fundamental living bed, which is known as Bio Ball!!
 
Nitrifying bacteria tends to be attached to porous surface fixture, if placed in the pool water for its attachment, it can be quickly attached to the fixture surface and begin to proliferate. They are largely non-motile and must colonize a surface (gravel, sand, synthetic biomedia, etc.) for optimum growth. They secrete a sticky slime matrix which they use to attach themselves.

However, the placement of the fixtures in the pool water is usually not feasible, the reason is that this way may hinder the activities of the fish and is not conducive to fishing pond. More feasible approach is to place a bio ball in a external filtration system, act as breeding bed for nitrifying bacteria.

Greatest Microbes of all time in Aquaculture - Bacillus Subtillis Natto

Probiotics in aquaculture is not a new thing now a day. However, many probiotics product being sold to aquaculture industry are not in fact true probiotic and may be even killing if it is not used accordingly. For example, some strain of soil bacteria do not have a recognized approval for food-grade probiotic use, but being used to feed the aqua animal. That’s way it is very important to understand the concepts of probiotics and their application in aquaculture.

When I started to work with probiotics in aquaculture, there were still lots of local farmer didn’t understand the concept of probiotic bacteria.

Mis-conception1
People simply put all the nutrient ingredient, (soy meal, fruits waste, molasses) and let them fermented or brewed for some period of time, after that just pour the fermented liquid into the pond, They thought these steps are simple and low cost, estimated the fish will grow better. Of course, the fermented liquid will have certain ‘effect’ on the pond, whether positively or negatively, but the outcome always inconsistent. They miss out an important concept, which is to enrich the important and selected strain of microbes; instead, they were enriching a very diversified and some ‘unfriendly’ microbes to the pond, which is not a wise idea to improve the quality of the water.

Mis-conception 2
Many shrimp and fish farmers often think of probiotic as fast-acting medicine like antibiotic. They expect a quick and instant effect. They are then discouraged when they see no instant and positive effects. In fact, the changing of bacterial community takes time. It is an ongoing process that requires the addition of the selected strain of bacteria throughout the culture period. The bacteria that added must be selected for specific function, added in high population density, under right environment to be effective!
Well, the term probiotics has been defined as “ a mono or mixed culture of living microbes that when applied to animal or man, affect beneficially the host by improving the properties of indigenious micro flora. So, to extend the definition for aquaculture to include the addition of natural bacteria to tank or pond in which the animal lives.

To date, Bacillus subtilis (natto strain) are recently gaining more and more popular in aquaculture industry. This bacteria are not harmful to aquatic animal and has been recognized safe to eat for living animal. They are actually occur naturally in the intestine tract of sea and fresh water animal.

In particular condition, certain strain of bacillus subtilis can even have better and effective role than conventional phototropic bacteria for aquaculture, because of their high tolerance towards stress environment (pH, salinity, temperature..) , denitrifying and hydrolyzing organic residue power.  These beneficial bacteria improve the health of fish or shrimp by suppressing the pathogen, by releasing natural antagonist antibiotic, which are not only selectively against the pathogenic microbes like. Besides, they have a very fast reproduction time, enabling them to easily become the dominant microbes in the pond, hence, improving water quality by modifying the microbial community composition of the water and sediment.
Ineffective commercialized probiotic some contain inappropriate species of bacteria, low population density. For example, some land animal probiotics that contain lactobacillus sp, are not appropriate for aquatic animal, as these bacteria do not occur originally in fish intestine.

Well, I am not ‘criticizing’ the lactobacillus product, just the we have to understand that the microbes can only function well under right environment. For example, in certain condition, , oxygen is easily depleted as the feeding rate was high, we have to clear off the accumulating faeces or food residue as soon as possible, to avoid excessive ammonia production and toxin excretion from vibrio species as a result of residue fermentation, aeration only is not enough, that is the time that we need to introduce high proteolytic and lipolytic ( the ability to break down large protein and fat molecule) activity bacillus subtilis where it will be very efficient to break down the residue and contribute to ammonia denitrifying. Whereas lactobacillus is a facultative anaerobic and low proteolytic activity bacteria ( live without oxygen) , the condition will not favors their growth and eventually they will become ‘malfunctioning’.

Therefore, if we want to become successful in aquaculture farming, we have to master the functioning concept and application of various microbes

However, bacillus subtilis are only cost- effective when that are apply in appropriate method with a suitable farm management.

General use/application of Bacillus Subtilis

I got it - Bacillus Subtilis!!

The time I got to know it because my friends invite me to go for a Japanese food dinner and introduce the Natto bean to me, a sticky and ammonia-odor smell, which date back few years ago. From the experience of natto , I know that this bacillus subtilis are simply awesome. People manipulate it on probiotic food, aquaculture, agriculture, household and septic system.

My Natto Experiment:
After few times of trying and trying... eventually successfully making a 'sticky' and 'ammonia-like' natto bean:)

The natto self-designed incubator (polyethylene box), which i maintain the inside-temperature 40 Celsius for bacillus subtilis optimum growth

General application

Bacillus subtilis are attractive industrial organisms for a variety of reasons, including their high growth rates leading to short fermentation cycle times, their capacity to secrete proteins into the extracellular medium, and the GRAS (generally regarded as safe) status with the Food and Drug Administration. The application of bacillus fermentation in food, biopharmaceuticals, enzyme industry, agriculture, aquaculture has been widely recognized and commercialized. For instances,

In addition to its role as model organism, Bacillus Subtilis is used in

1. Commercial production of Natto Bean
2. Soil Inoculant for agricultural farming
3. Biological control agent.
4. As an immunostimulatory agent, probiotic to aid treatment of gastrointestinal and urinary tract diseases.
5. It can convert some explosives into harmless compounds of nitrogen, carbon dioxide, and water.
6. Production of biosurfactant which having antibacterial effect apply in medical tools.
7. Production of various enzymes protein, such as amylase, hyaluronic acid.

 

Saw-dust /chicken dung organic fertilizer that inoculated with Bacillus Subtilis

Bacillus Subtilis ( the middle white spot), which i isolated from organic fertilizer (picture above)

Nowadays, in prospect of biochemistry, physiology, and genetics of B. subtilis, more and further development and exploitation are in advanced growth. The complete genome for B. subtilis 168 was recently published. With this great achievement in applied and industry microbiology, enable the scientist to produce new beneficial genetic-engineered products.

If u want to know more how this microbes functioning, please do google for that, I am you will be amazed by their works.

There are thousands of journals characterizing this microbe of interest, and it is impossible for me to introduce them in detailed way. Hence, I try to make a brief introduction of this particular microbe.

History ( From Wikipedia)

In 1835, the bacterium was originally named Vibrio subtilis by Christian Gottfried Ehrenberg, and renamed Bacillus subtilis by Ferdinand Cohn in 1872. Cultures of B. subtilis were used throughout the 1950s as an alternative medicine due to the immunostimulatory effects of its cell matter, which upon digestion has been found to significantly stimulate broad spectrum immune activity including activation of specific antibody IgM, IgG and IgA secretion and release of CpG dinucleotides inducing INF A/Y producing activity of leukocytes and cytokines important in the development of cytotoxicity towards tumor cells.

It was marketed throughout America and Europe from 1946 as an immunostimulatory aid in the treatment of gut and urinary tract diseases such as Rotavirus and Shigella, but declined in popularity after the introduction of cheap consumer antibiotics, despite causing less chance of allergic reaction and significantly lower toxicity to normal gut flora.

Morphologic and biochemistry

They are Gram-positive, catalase-positive bacterium, rod-shaped, and has the ability to form a tough, protective endospore, allowing the organism to tolerate extreme environmental conditions.

Reproduction

B. subtilis can divide from a single cell to make two daughter cells, within 30 min with favorable condition. Amazing right!!  In addition, it forms single endospore that can remain viable for decades and is resistant to unfavourable environmental conditions such as drought, salinity, extreme pH, radiation and solvents. The endospore is formed at times of nutritional stress, allowing the organism to persist in the environment until conditions become favorable. Most of the commercial product, agriculture and aquaculture grade are in spore-form condition, which mean they are simply in sleeping mode, covering them with a protective layer. Because of this characteristic of resistance and stability, people are happy to commercialize it into product, where they can keep it for long period.

Prior to the process of sporulation the cells might become motile by producing flagella, take up DNA from the environment, or produce antibiotics. These responses are viewed as attempts to seek out nutrients by seeking a more favourable environment, enabling the cell to make use of new beneficial genetic material or simply by killing of competition. Wild-type natural isolates of B. subtilis are difficult to work with compared to laboratory strains that have undergone domestication processes of mutagenesis and selection. These strains often have improved capabilities of transformation (uptake and integration of environmental DNA), growth, and loss of abilities needed "in the wild." And, while dozens of different strains fitting this description exist, the strain designated 168 is the most widely used.