Amendments to the Enforcement Ordinance of the Standards of Feed and Feed Additives  
  
The government of Japan will  add standard s and specification s of Bacillus subtilis  and Phytase .  
Major additions  are outlined  below.  
 
 
Bacillus subtilis No.5  
 
Specifications for feed additives  
Active substance  
Compositional specifications  
This active substance is a lyophilized or frozen  Bacillus subtilis JA-ZK strain intended for the 
use in manufacturing.  
Origin   The original strain is the Bacillus subtilis JA-ZK strain isolated from soil in 2000.  
Methods of preservation and criteria for passage  
Passage the original strain in culture media containing nutrients such as yeast extract and starc h. 
Preserve the original strain at ≤ -70 °C  frozen , or at 2 ~ 8 °C after lyophilized . 
Grow the original strain using the same media and store the lyophilized or frozen active 
substance in small quantity. The maximum number of passage of the original strain  is 10. This 
product itself should not be used for passage.  
 
Product  
Compositional specifications  
This product is in the form of powder. This product is made from mixing bacterial cells 
collected after cultivation of Bacillus subtilis  No.5 active substanc e and starch, then dry and mix 
excipient substance.  
Content    1 g of this product contains 10-1 times to 10-2 times of live bacteria corresponding to 
the content on the label.  
Standards for methods of manufacture  
Culture the original strain of  Bacillus subtilis  No.5. Collect bacterial cells, add starch, dry and 
mix excipient substance.  
Standards for methods of storage  
Store the products in airtight containers.  
 
Specifications for methods of manufacture in general  
This Bacillus subtilis  is allow ed to be used in feed for broiler, layer and pig. 
 
 
 Phytase  No. 2-3 
 
Specifications for feed  additives  
Active substance  
Compositional specifications  
Unit     ≥ 5000FTU/1g   
Physical and chemical properties     
1. This  active substance is light brown liquid and has characteristic odor . 
2. pH of aqueous solution or aqueous  suspension  (1→100) is 3.5  to 6.5. 
3. This active substance  has the best enzyme activity  when pH is between pH4.5 and pH6.0.  
Ignition residue  ≤ 5.0% (0.5g)  
Standards for methods of manufacture  
Culture  genetically modified Schizosaccharomyces pombe . Filter the preparation or extract it 
with water . Then, concentrate the filtrate.  
 
Product  1 
Compositional specifications  
This product  is in the form of liquid and is made from mixing sodium chloride , citric acid and 
sorbitol  to the a ctive substance  of Phytase No . 2-3. 
Unit     This product  contains Phytase  correspond ing to 85 to 170% of the FTU on the label . 
 
Product 2 
Compositional specifications  
This product is  in the form of pieces, powder or particle s and is made from mixing or 
granulating after adding sodium sulfate , excipient substances , and a queous solution of polyvinyl 
alcohol  if necessary, or  piece s, powder or particle s made from  mixing  citric acid, flour and 
calcium prop ionate . 
Unit    This product  contains Phytase correspond ing to 85 to 170% of the FTU on the label . 
 
Specifications for methods of manufacture in general  
This Phytase  is allowed to be used in feed  for broiler , layer  and pig.