| Bacillus coagulans : A
superior probiotic
L. sporogenes* was first isolated and described in
1933 by L.M. Horowitz-Wlassowa and N.W. Nowotelnow and the name was accepted in the fifth
edition of " Bergeys Manual of Determinative Bacteriology". However, it
was transferred to Bacillus coagulans in the seventh edition of Bergeys
manual due to simplification in cataloging. However, in honor of the original discoverers,
the name Lactobacillus sporogenes* is used widely, except for taxonomical
purposes. According to the Eighth Edition of Bergeys Manual of Determinative
Bacteriology, "Various spore-bearing rods which produce lactic
acid, are facultative or aerobic and catalase positive, have generally and correctly been
assigned to the genus Bacillus."
The characteristics
of L. sporogenes* as cited in Bergeys Manual (Seventh Edition) and other
sources are: "Gram - positive spore-forming rods 0.9 by 3.0 to 5.0 micron size,
aerobic to microaerophilic, producing L(+)-(dextrorotatory) lactic acid
homofermentatively." Since L. sporogenes* exhibits characteristics typical of
both genera Lactobacillus and Bacillus, its taxonomic position between the
families Lactobacillaceae and Bacillaceae has often been discussed. This,
along with the fact that there is no universally accepted official classification leaves
room for controversy in the nomenclature. The differentiation
characteristics of L. sporogenes* are indicated in Table 2.4:

Table 2.4 : L. sporogenes* -
key characteristics for differentiation:
Property Bacillus Lactobacillus Sporolactobacillus L. sporogenes*
It is often very
difficult to distinguish between two species of bacteria which are morphologically similar
and possess similar physiological and biochemical characteristics. DNA-DNA homology is an
useful technique in resolving this difficulty. The nucleotide sequence in the DNA of
various bacteria differs. Additionally, DNA from closely related bacteria bind to each
other more efficiently. These facts are used in the typing and classification of bacteria
and have been effectively employed to recognize the innate resemblance between L.
sporogenes* and members of the genus Lactobacillus. The results of a detailed
study prove conclusively that L. sporogenes* is closer to Lactobacillus
than to Bacillus, irrespective of its taxonomical placement under genus Bacillus in the Bergeys manual.
The characteristics
by which L. sporogenes* resembles the genus Bacillus are as listed:
Cells are long
and slender (0.3 to 0.8 mM), some are bent and all the cells have rounded ends.
Motile with
peritrichous flagellas
Gram positive
Colonies are
usually 2.5 mm in diameter, convex, smooth, glistening and do not produce any pigment.
Extremely
fastidious organisms requiring complex organic substrates for growth such as fermentable
carbohydrate, peptone, meat and yeast extract. MRS medium supplemented with tomato juice,
manganese, acetate and Tween-80 is a suitable medium for growth.
Grow optimally at
30° C to 37° C and the optimum pH in the range 5.5 to 6.2.
Microaerophilic,
exhibit fermentative metabolism and are facultatively aerobic.
Produce acid from
arabinose, xylose, glucose, galactose, mannose, fructose, maltose, sucrose and trehalose.
Do not hydrolyse
starch or casein.
Do not liquefy
gelatin
Are indole
negative and do not produce hydrogen sulfide or gas.
Produce
dextro-rotatory lactic acid from glucose, fructose, sucrose, trehalose and inulin.
Menaquinones are
absent.
The characteristics
by which L. sporogenes* differs from the genus Bacillus are as listed:
Species of
Bacillus are always rod-shaped and straight but lactic acid bacteria also appear as bent
rods and exhibit very little motility.
The spores
present are terminal in position and are not centrally located as in Bacillus.
Lactic Acid
Bacteria are oxidase negative (OXIDASE TEST)
Do not reduce
nitrate to nitrite while most Bacilli reduce nitrate.
The mole % G+C of
DNA in Bacillus ranges from 32 to 69% whereas in Lactobacillus it does not exceed 53%.
L. sporogenes* was assigned to genus Lactobacillus based on the above characteristics by Nakayama
and Yanoshi (1967).
L. sporogenes* grows
in the temperature range of 35° C to 50° C; the optimum pH range is 5.5-6.5. Unlike
other lactobacilli currently in clinical use, L. sporogenes* can form spores.
Sporulation is the development in microorganisms of bodies each wrapped in a protective
coat (a natural process of microencapsulation in a calcium-dipicolinic acid- peptidoglycan
complex). Under favorable conditions, the spores germinate into viable bacilli and carry
on their life activities. The spores of L.sporogenes* are ellipsoidal bodies
measuring 0.9 to 1.2 by 1.0 to 1.7 microns. Their morphology and formation are
schematically represented in Figures 2.6 and 2.7:


This property of
spore formation by L. sporogenes* is the main characteristic that makes it the
probiotic of choice in clinical applications. On oral administration, these spores survive
the acidic gastric environment and are activated due to the low pH, mechanical churning
action of the stomach and the water in the gastric environment. The spore coats imbibe
water, swell, and the increased water content causes a rise in the metabolic rate of the
sporulated bacilli. Outgrowths begin to protrude from the spore-coats. The spores pass on
to the duodenum where the outgrown cells germinate and transform into viable vegetative
cells. They begin to proliferate in the small intestine, multiplying rapidly. Usually,
germination takes place about four hours after ingestion. A large supply of viable L.
sporogenes* is thereby ensured in the small intestine. These cells settle in the
intestinal tract and continue their metabolic activities, producing lactic acid and
probably bacteriocins, which render the intestinal environment non-conducive for the
growth of harmful pathogenic bacteria. The maintenance of a low,
constant level of lactic acid on the inner surface of the intestinal tract helps restore
the microecological balance after antibiotic therapy. Antibiotic therapy may kill
beneficial microbes which help in the synthesis of vitamins B and digestive enzymes. Since
L. sporogenes* produces only L (+)-lactic acid, it
does not cause metabolic acidosis.
Morphological and physiological
characteristics of L. sporogenes*
The vegetative
cells are rods occurring singly, rarely in short chains, the filaments varying with
cultural conditions. The cells are motile by means of flagella.
Carbohydrate fermentation:
| Carbohydrate |
Acid
production |
Gas
production |
| Inulin |
- |
- |
| Maltose |
+ |
- |
| Mannitol |
+ |
- |
| Raffinose |
+ |
- |
| Sorbitol |
- |
- |
| Sucrose |
+ |
- |
| Trehalose |
+ |
- |
Properties of the spores:
The spores of L.
sporogenes*74 are resistant to heat and other adverse environmental conditions,
surviving even under a temperature of 100° C for twenty minutes in phosphate buffer at pH
7. The spores germinate in malt broth even in the presence of dilute hydrochloric acid (at
pH 4.6 to 5.6), caustic soda solution (pH 7.6-9.6), saline solution (5%, 10%, 20%
concentration), a 2.5% solution of boric acid as well as distilled water. The spores are
two to eight times more resistant to antibiotics than the vegetative cells. |