Probiolac

STABILITY TEST REPORT

1.00E+121.00E+111.00E+101.00E+091.00E+08CFU / Gm 1.00E+071.00E+061.00E+051.00E+041.00E+031.00E+021.00E+011.00E+00-

April 2007

July 2008

April 2009

July 2009

April 2010

April 2011

Mar 2008

Jan 2008

Jan 2009

July 2010

Date / Period Product Count

Comment : The Sample continues to demonstrate excellent stability. Counts remain in the 10th power range. Place : Salem Date : 30.10.11 Respectfully Submitted

L.Poonmani Microbiologist

July 2011

Jan 2010

Oct 2007

Oct 2009

Nov 2008

Jan 2007

Jan 2011

Oct 2010

Jul 2007

M/s.Intvet Products Mfg.Co.(P)Ltd., 171-G, Rajapuram Nedunchalai Nagar Salem.5

Report : #3-333-707 Subject : Heat/Time study

Purpose :
To compare the Heat stability of three products at 190 F. to simulate the temperature of peleting.

Procedure :
Five grams of material were placed in a sterile glass bottle, heated to 190 F. This bottle was placed on its side and rotated to spread material out to get the largest surface area possible to expose to the heat. The bottle was placed on the side in the over. Samples were drawn as rapidly as possible the following three time points: (15 minutes), (30 minutes) and (45 minutes) in addition & To sample was taken on a reference and a control. The samples were placed in a sterile glass bottle with 100ml distilled water and shaken for two hours on a rotary shaker followed by blending two minutes at high speed. A terifold serial dilution series was set up and applicable dilutions were placed using MRS Lactobacillus Agar.

Results :
The average temperature throughout the experiment was 189.8 F. A comparison of stability of PRODUCT -X, PROBIOLAC and PRODUCT -Y 10 at 188.8 F. for a verified time. Lactobacillus sp bacteria were enumerated using NRIA procedure. Sample Product Product -X Probiolac Time colony forming TO 1,000 1,300 T-15 200 1,300 400 Units / gram T-30 0 1,100 800 T-45 0 900 300 %Loss of viability at 45 minutes 100% 30.8% 99.9%

Product -Y 108,000,00

Both probiotics and Product -Y 10 rapidly loose viability at 10.88.8F. Probiolac appears to be fairly stable at 1.88.8 F. showing approximately 31% loss at 48 minutes. The small increase in counts in the Product -Y 10 material at T-30, is probably a sampling error. To try to minimize the temperature various the samples were weighed rapidly, this could account for the discrepancy.

CONCLUSION :
Probiolac is a more stable product at 188.8 F. than the other two products tested. The decrease in the Probiolac counts seen above and not in any other tests, is due to the fact that prolonged temperature (In excess of 15 minutes ) is pushing the limit of even what your product can withstand.

M/s.Intvet Products Mfg.Co.(P)Ltd., 171-G, Rajapuram Nedunchalai Nagar Salem.636 005

Report : #3-0276-702

Dear Sirs, Here are the results of a study we conducted using Intvet Products Probiolac, animal culture, Lot No. 2293-16. The purpose was to do a time course study to determine the number of lactobacillus sp. surviving at 185 F. This temperature was chosen to approximate the temperature of pelleting. The time points chosen were 0.15, 30, 45 and 80 minutes. For the experiment, no other materials were added to the Lactobacillus animal culture. The counts were made using the N.F.I.A method which I feel is more sensitive than other methods. Time at 155.5 0 min. 15 min. 30 min. 45 min. 80 min. Lactobacillus / gram 2.7 X 10 6 2.1 X 10 6 2.5 X 10 6 2.0 X 10 6 3.1 X 10 6

The data shows significant loss of viability through one hour at 185 F. The slight increase in bacterial numbers at one hour is most likely due to a sampling error. The bottom line is, that the Intvet Products Lactobacillus will survive the high temperatures encountered in pelleting, and in fact, will survive for periods of time much longer than normal pelleting time.
Respectfully submitted

L.Poonmani Microbiologist

M/s.Intvet Products Mfg.Co.(P)Ltd., 171-G, Rajapuram Nedunchalai Nagar Salem.636 005 Subject : Lactobacillus Analysis Sample Identification Probiolac Lactobacillus count Time course 90 F Day 0 Day 7 1.2 X 107 CFU/g 1.25 X 107 CFU/g

Laboratory Number 1777-1

1777-2

Product -X

1.0 X 109 CFU/g

2.0 X 105 CFU/g

1777-3 Method

Product -Y XFIA

4.3 X 105 CFU/g Sensitivity of method

8.5 X 104 CFU/g 100 CFU/g

Comment : All methods used are XFIA approved. The various products were stored in tightly sealed containers at 90 for seven days. Product Probiolac Product -X Product -Y Respectfully Submitted L.Poonmani Microbiologist % loss No loss 80% loss 100% (effective loss)