Field of invention
The invenlion disclosed in this application relates to an improved composition (Medium) which is useful for assay of a probiolic and a process for its preparation
Probiotics are simple bacteria or a mixture of microoruanisms which have been mutated or rendered non pathogenic. They are administered m oral form to patients who may have disturbances in the normal microbiota of the gastro intestinal tract due to infections or antibiotic therapy. Probiotics are also used to treat infections in place of antibiotics and hence have been postulated to have inhibitatory effects on the growth of pathogenic microorganisms.
The action of probiotics is studied in vitro by using bacteriological media similar to those used to study antibiotic susceptibility such as Mueller Hmton agar broth which has low content of thymidine and cations both of which are known to decrease the activity of some antibiotics. Such a medium however is not ideal to test the activity of a probiolic which consists of a mixture of organisms all of which may have different growth requirements Meat particles are known to produce a low O/R (oxidation reduction potential) thai facilitales the growth of anaerobes.
Prior art
Bacteriological media containing meat particles are commercially available and are used to cultnate facuhatne and strict anaerobes Such a medium however is not used ibr antibiotic assays since it is not free of all substances that may be inhibiton to some antibiotics- Mueller Hinton medium broth has been used to assay probiotics and antibiotics: however it supports the growth of only aerobes and facultative anaerobes and may not be ideal if the probiotic consists of a mixture of organisms of different oxygen requirements.


Still another objective of the present invention is to provide a process for the preparation ot an miprosed composition (medium) which will support the growth of different types of probiotic organisms.
The composition ( medium ) of the present invention can support the growth of different kinds of organisms of varied oxygen and nutritional requirements and is also suitable for probiotic and antibiotic assays. As mentioned above there is no


Sterfle probiotic assay medium when inoculated with mixed probiolic organisms- ailow the growth ofaerobes in the broth and the growth of anaerobes on the meal particles After an incubation period of 24-72 hours at 37° C in an atmosphere of about 7-10 % Co? the antimicrobial acti\ ity can be studied using the supernatant that contains the metabolites against the pathogens of interest.


be incubated at 37°C for 72 hours. The cell free supernatant can be obtained by cenlnfugalion at 5000 rpm and it's antimicrobial efficacy can be tested against broth cuhures of the pathogens of interest.

AS PER THE PROCESS DEFINED ALL THE ABOVE INGREDIENTS ARE MIXED & AUTOCLAVED TOGETHER . BUT ACCORDING TO THE PROCESS DESCRIBED IN THE EXAMPLES ONLY MINCED MEAT/ BEEF HEART IS AUTOCLAVED AND OTHER INGREDIENTS ARE ADDED AFTER AUTOCLAVING THEREFORE THESE TWO PROCESSES ARE INCONSISTENT THEY SHOULD BE MADE CONSISTENT


Since each organism is able to grow optimally and metabolise, the metabolic by products accumulate in the supernatant, which can easily be assayed. Toxic by products that may inhibit the metabolites are adsorbed by certain components (starch) of the medium. Hence probiotic organisms grow well.

We Claim
1. An improved composition (media) useful for probiolic assay which comprises (i) Meal pieces in an amount in the range of 5 to 80 % b\ wl ((h) Peptone in an amount in the range of 0,5 to 10% by \\t (iii) Sugar in an amounta in the range of 0.1 to 20.0% by wt (i\) Salt (Na CI) in an amount in the range of 0.1 to 10.0 % by wt (v) Protein tndrolysate in an amount in the range of 0.1 to 10% by wt (vi) Meal infusion in an amount in the range of 10 to 90% by wt and (\ii) Starch in an amount in the range of 0.05 to 10.0 % by wt. the pH of the resulting composition ( media ) being in the range of 7.4 + / - 0.2
2 An improved composition (media) as claimed in claim 1 wherein the amount of Meat pieces used ranges from 30 to 50% by wt
3. An improved composition (media) as claimed in claims 1 & 2 wherein the amount of Proteose peptone used ranging from I to 3 % by wt
4. An improved composition (media) as claimed in claims 1 to 3 wherein the amount of Dextrose used ranging from i to 2 % by wA
5 An improved composition (media) as claimed in claims 1 to 4 wherein the amount of NaCI used ranging from 0.3 to 0.6 % by wt
0. An improved composition (media) as claimed in claims 1 to 5 wherein the amount of Casein hydrolysate used ranging from I to 2 % by wt
7. An improved composition (media) as claimed in claims 1 to 6 wherein the amount of Beef infusion used ranging from 25 to 45 % by wt

8. An improved composition (media) as claimed in claims 1 to 8 wherein the amount of Soluble starch used ranging from -0.1 to 0.2 % b\ \\l.
9, An improved composition (media) as claimed in claims 1 to 8 wherein the pH
of the composition { media) is maintained at 7.5
10. A process for the preparation of an improved composition (media) as claimed
in claims 1 to 8 which comprises auloclaving the mixture of (i) Meat pieces in an
amount in the range of 15 to 50 % by wt ((ii) Proteose peptone in an amount in the
range of 1 to 5% by wt (iii) Dextrose in an amount in the range of 0,5 to 2.0% by
wt (i\) NaCl in an amount in the range of 0.2 to 0.7 % by wt (v) Casein
hydrolysate in an amount in the range of 0.5 to 2% by wt (vi) Beef infusion in an
amount in the range of 20 to 50% by wt and (vii) Soluble starch in an amount in
the range of 0. i to 0.2% by wt. at a temperature in the range of 115o C - 135o C. at
a pressure m the range of 101bs to 30 lbs of pressure for a period in the range of 15
to 30 minutes- cooling the resulting composition (medium) to room temperature
1 1 A process as claimed in claim 10 wherein the autoclaving is effected at a temperature in the range of 120o C to 135oC at a pressure in the range of 10 to 15 lbs of pressure for a period in the range of 15 to 20 minutes .
12. A process as claimed in claims 10& 11 wherein the amount of Meat pieces used ranges from 30 to 50% b> wl
13. A process as claimed m claims 10 to 12 wherein the amount of Proteose peptone used ranging from 1 to 3 % by wt
!4. A process as claimed in claims 10 to 13 wherein the amount of Dextrose used ranging from 1 to 2 % by wt
15 A process as claimed in claims 10 to 14 wherein the amount of NaCl used ranging from 0.3 to 0.6 % by wt

K), A process as claimed in claims 10 to 15 wherein the amount ol Casein hydroly sale used ranging from 1 to 2 % by wt
17 A process as claimed in claims 10 to 16 wherein the amount of Beef infusion
used ranging from 25 to 45 % by w t
1