Guidance for FDA Staff 
 
Compliance Policy Guide 
Sec. 527.300 Dairy Products - 
Microbial Contaminants and Alkaline 
Phosphatase Activity  
 
 
Additional copies are available from: 
Division of Compliance Policy HFC-230 
Office of Enforcement 
Office of Regulatory Affairs 
Food and Drug Administration 
5600 Fishers Lane 
Rockville, MD  20857 
(Tel) 240-632-6860 
http://www.fda.gov/ora/compliance_ref/cpg/default.htm 
 
You may submit either electronic or written comments regarding this guidance at any 
time.  Submit electronic comments to http://www.regulations.gov.  Submit written 
comments to the Division of Dockets Management (HFA-305), Food and Drug 
Administration, 5630 Fishers Lane, rm. 1061, Rockville, MD  20852.  All comments 
should be identified with the docket number listed in the notice of availability that 
publishes in the Federal Register. 
  
 
U.S. Department of Health and Human Services 
Food and Drug Administration 
Center for Food Safety and Applied Nutrition  
Office of Regulatory Affairs  
 
December 2010  Contains Nonbinding Recommendations 
Table of Contents 
 
 
  I.  Introduction 
 
 II.  Background 
 
III.  Policy 
 
IV.  Regulatory Action Guidance 
 
 V.  Specimen Charges  
  2 Contains Nonbinding Recommendations 
Guidance for FDA Staff 
 
Compliance Policy Guide 
Sec. 527.300 Dairy Products - Microbial Contaminants 
and Alkaline Phosphatase Activity 
 
This guidance represents the Food and Drug Administration's (FDA's) current thinking on 
this topic.  It does not create or confer any rights for or on any person and does not 
operate to bind FDA or the public.  You can use an alternative approach if the approach 
satisfies the requirements of the applicable statutes and regulations.  If you want to 
discuss an alternative approach, contact the FDA staff responsible for implementing this 
guidance.  If you cannot identify the appropriate FDA staff, call the telephone number 
listed on the title page of this guidance. 
 
I.  Introduction: 
 
*The purpose of this document is to provide guidance for FDA staff regarding its 
enforcement policies for pathogens and indicators of inadequate pasteurization or post-
pasteurization contamination of dairy products. 
 
FDA's guidance documents, including this guidance, do not establish legally enforceable 
responsibilities.  Instead, guidances describe the Agency's current thinking on a topic and 
should be viewed only as recommendations, unless specific regulatory or statutory 
requirements are cited.  The use of the word should in Agency guidances means that 
something is suggested or recommended, but not required.  
 
II.  Background: 
 
Significant outbreaks of illness associated with dairy products include salmonellosis 
(including infections with some multi-drug resistant Salmonella), enterohemorrhagic 
Escherichia coli (EHEC) O157:H7 infections, listeriosis, staphylococcal food poisoning, 
botulism, and Yersinia enterocolitica infection.  (A Federal Register notice of availability 
for draft CPG Sec. 555.320 Listeria monocytogenes was published on February 7, 2008 
(73 FR 7293)).  Symptoms of illnesses from pathogens may include anything from mild 
discomfort to headaches, nausea, vomiting, watery or bloody diarrhea, cramps, and 
hemolytic uremic syndrome.  In some cases, death may result.   
 
A. Pathogens 
The presence of pathogens in dairy products is an indicator of poor sanitation, 
temperature abuse, inadequate pasteurization (21 CFR 133.3(d)), fermentation failure, or 
obtaining milk from diseased animals.  Many pathogens originate from the feces of the 
  3 Contains Nonbinding Recommendations 
dairy animals themselves.  Contamination of the udder and teats by the feces may in turn 
contaminate raw milk.  Insanitary practices by milk handlers may contribute to the 
contamination.  Insanitary or inadequate processing conditions also may contaminate the 
raw milk dairy product.  Pasteurization of raw milk is lethal to pathogens. Thus, the 
presence of pathogens in a dairy product made from milk that was subjected to a 
pasteurization process indicates inadequate pasteurization or post-pasteurization 
contamination. 
 
Salmonella - Salmonella is a pathogen that, when consumed, can cause an infection.  A 
dose of as little as 15-20 organisms can cause illness.  The symptoms of infection include 
gastroenteritis.  Salmonella is shed in the feces of infected animals and can contaminate 
pasture land and milking parlors.   
 
EHEC O157:H7 and other enterohemorrhagic Escherichia coli - The infectious dose 
of EHEC 0157:H7 is estimated to be between 10-100 organisms. When food 
contaminated with the EHEC O157:H7 is consumed, the pathogen colonizes the intestinal 
tract where it produces a toxin and causes haemorrhagic colitis that can progress to more 
serious complications such as haemolytic uraemic syndrome or thrombotic 
thrombocytopenic purpura.  EHEC O157:H7 is the predominant EHEC strain that has 
caused illness worldwide.  However, other EHEC serotypes have also been implicated in 
illness and are of public health concern.  
 
Campylobacter jejuni - A dose of 400-500 organisms of Campylobacter jejuni can cause 
infection.  Symptoms of infection include abdominal pain, fever, diarrhea, and vomiting.   
Campylobacter jejuni can either be shed in feces or in milk from an infected udder of a 
dairy animal.  Most human outbreaks of infection of Campylobacter jejuni that are 
associated with dairy products have been linked to raw milk or inadequately pasteurized 
milk.    
 
Yersinia enterocolitica - Yersinia enterocolitica is a pathogen which causes infection.  
Symptoms of infection include gastroenteritis, fever, diarrhea, bloody stools, rash, joint 
pain, nausea, vomiting, headache, and malaise.  Infection by Yersinia enterocolitica is 
also considered a cause of reactive arthritis.  Yersinia enterocolitica has been found in 
many different animals and is shed in feces. 
 
Clostridium botulinum - Clostridium botulinum produces a neurotoxin.  A few 
nanograms (ng) of the neurotoxin can cause illness.  Symptoms include lassitude, 
weakness, vertigo, double-vision, difficulty speaking, and difficulty swallowing.  
Symptoms can progress to difficulty of breathing, weakness of other muscles, abdominal 
distention, and constipation.  The incidence of this disease is low, but the mortality rate is 
high if not treated immediately and properly.  Although the neurotoxin is heat labile and 
can be destroyed when exposed to a minimum of 80 oC for 10 minutes, the time and 
temperature reached in pasteurization do not destroy the neurotoxin.  
 
Enterotoxigenic Staphylococcus  Some species of Staphylococcus produce an 
enterotoxin that is extremely heat stable and is not inactivated at pasteurization 
  4 Contains Nonbinding Recommendations 
temperatures.  When ingested, the enterotoxin may rapidly produce symptoms including 
nausea, vomiting, retching, abdominal cramps, muscle cramping, headache, and transient 
changes in blood pressure and pulse rate.  The presence of any Staphylococcus 
enterotoxin in a dairy product is of public health concern. 
 
A dairy cow with mastitis may be the source of enterotoxigenic Staphylococcus in raw 
milk, which may subsequently be commingled with other milk.  Also, at any point from 
the milk collection process to the packaging of the finished product, enterotoxigenic 
Staphylococcus species can be introduced by an infected human, inadequate employee 
hygienic practices, such as inadequate hand washing, equipment and utensils that are not 
cleaned and sanitized, or contaminated raw material. 
 
Staphylococcus aureus has traditionally been used as a microbiological indicator of 
insanitation during processing.  Because of environmental factors, low levels of 
Staphylococcus aureus may be found in raw milk, even when produced using good 
manufacturing practices (GMPs).  However, excessive numbers of Staphylococcus 
aureus organisms in raw milk or other dairy products, i.e., greater than or equal to 104 
cfu/g, indicate that the product was produced under insanitary conditions. 
 
Bacillus cereus - Bacillus cereus can cause illness when 106 cfu/g or more are consumed 
in food.  There have been two enterotoxins produced by Bacillus cereus identified as 
causing foodborne illness.  Illness is characterized by abdominal pain and diarrhea or 
nausea and vomiting.  Bacillus cereus is commonly found in soil, on vegetables, and in 
many raw and processed foods, including milk and cheese. 
 
Additional information about foodborne pathogens and associated diseases can be found 
in the Bad Bug Book. 
 
B. Nontoxigenic Escherichia coli  
Escherichia coli  has traditionally been used as a microbiological indicator of insanitation 
during processing.  Escherichia coli are not inherently present in the milk of a dairy 
animal.  Escherichia coli in milk and dairy products generally originate from animal or 
human feces. Thus, the presence of Escherichia coli in milk or other dairy product means 
that the milk or dairy product was exposed either directly or indirectly to feces.  
 
Because of the close association of raw milk with the animal environment, low levels of 
Escherichia coli may be present in raw milk or products made from raw milk, even when 
properly produced using GMPs.  Insanitary conditions, including poor employee hygiene 
practices, improperly sanitized utensils and equipment, or contaminated raw materials, 
may also be a source of Escherichia coli in milk and other dairy products. 
 
C. Alkaline phosphatase 
Alkaline phosphatase is an enzyme that is naturally found in milk.  Although the alkaline 
phosphatase in milk is more thermal resistant than nonsporeforming microorganisms, it is 
denatured by pasteurization temperatures.  Therefore, the presence of alkaline 
phosphatase indicates the milk was not properly pasteurized.  The alkaline phosphatase 
  5 Contains Nonbinding Recommendations 
level should be negligible in dairy products, other than certain cheese and cheese 
products.  
 
For some dairy products such as certain cheeses, microorganisms used during production 
produce alkaline phosphatase.  For some cheeses, the maximum level of alkaline is 
established in the standard of identity for the cheese (21 CFR part 133).  For other cheese 
and related cheese products required to be made from pasteurized milk, the alkaline 
phosphatase level should not be greater than 3 g/0.25 g (12 micrograms phenol 
equivalents per gram). 
 
III.  Policy: 
 
FDA will review the available evidence on a case-by-case basis to determine whether a 
dairy product is adulterated and, in doing so, will be guided but not bound by the 
following general statements of policy relating to the presence in those products of 
pathogens, nontoxigenic Escherichia coli, or alkaline phosphatase.  
 
A. Pathogens 
Salmonella species, EHEC O157:H7 and other enterohemorrhagic Escherichia coli, 
Campylobacter jejuni, Yersinia enterocolitica, vegetative cells of Clostridium 
botulinum, Clostridium botulinum toxin, Staphylococcus enterotoxin, or Bacillus 
cereus enterotoxin 
Dairy products may be considered adulterated within the meaning of section 402(a)(1) of 
the Federal Food, Drug, and Cosmetic Act (the Act) (21 U.S.C. 342 (a)(1)) in that they 
bear or contain a poisonous or deleterious substance which may render them injurious to 
health when Salmonella species, EHEC O157:H7 and other enterohemorrhagic 
Escherichia coli, Campylobacter jejuni, Yersinia enterocolitica, vegetative cells of 
Clostridium botulinum, Clostridium botulinim toxin, Staphylococcus enterotoxin, or 
Bacillus cereus enterotoxin are present.  
 
Staphylococcus aureus 
Dairy products may be considered adulterated within the meaning of section 402(a)(4) of 
the Act (21 U.S.C. 342(a)(4)), in that they have been prepared, packed, or held under 
insanitary conditions whereby they may have been rendered injurious to health when 
Staphylococcus aureus are present at levels greater than or equal to 104 cfu/g. 
 
Bacillus cereus 
Dairy products may be considered adulterated within the meaning of section 402(a)(4) of 
the Act (21 U.S.C. 342(a)(4)), in that they have been prepared, packed, or held under 
insanitary conditions whereby they may have been rendered injurious to health when 
Bacillus cereus are present at levels greater than or equal to 104 cfu/g. 
 
B. Nontoxigenic Escherichia coli  
Dairy products may be considered adulterated within the meaning of section 402(a)(4) of 
the Act (21 U.S.C. 342(a)(4)), in that they have been prepared, packed, or held under 
insanitary conditions whereby they may have become contaminated with filth when 
  6 Contains Nonbinding Recommendations 
Escherichia coli is found at levels greater than 10 MPN per gram in two or more 
subsamples or greater than 100 MPN per gram in one or more subsamples. 
 
C. Alkaline phosphatase 
Dairy products made from bovine milk may be considered adulterated within the 
meaning of section 402(a)(4) of the Act (21 U.S.C. 342 (a)(4)) when:  
  For cheese and related cheese products required to be made from pasteurized milk 
(21 CFR part 133), alkaline phosphatase levels are: 
o  Greater than 5 g/0.25 g (20 micrograms phenol equivalents per gram) in one 
or more subsamples for brick (21 CFR 133.108), semisoft (21 CFR 133.187), 
and semisoft part-skim cheeses (21 CFR 133.188);  
o  Greater than 4 g/0.25g (16 micrograms phenol equivalents per gram) in one 
or more subsamples for Limburger cheese (21 CFR 133.152); or  
o  Greater than 3 g/0.25 g (12 micrograms phenol equivalents per gram) in one 
or more subsamples for all other cheese and related cheese products.   
  For dairy products other than cheese and related cheese products, alkaline 
phosphatase level is greater than or equal to 2.0 micrograms phenol equivalents 
per gram in one or more subsamples.  
 
IV.  Regulatory Action Guidance:  
 
Interstate Milk Shippers 
 
Dairy products produced in an Interstate Milk Shippers (IMS) listed plant that may be 
considered adulterated under the Act should first be referred to the appropriate State 
regulatory agency for follow-up.  This includes a non-IMS product manufactured in an 
IMS-listed plant for which the State is providing a regulatory program to address these 
issues.  If the State is unable to take appropriate action, the district should proceed to 
initiate FDA action as indicated below.  
 
Direct Reference 
 
The following represent criteria for direct reference seizure submission to the Division of 
Compliance Management and Operations (HFC-210), for direct reference import 
detention by the district and for direct reference submission of detention without physical 
examination (DWPE) to ORA, Office of Regional Operations, Division of Import 
Operations and Policy (HFC-170): 
Analysis of the dairy product demonstrates that one or more subsamples is positive 
for Salmonella species, Campylobacter jejuni, Yersinia enterocolitica, 
Staphylococcus enterotoxin, or Bacillus cereus enterotoxin. 
 
Recommendations 
 
The following represent criteria for recommending seizure, import detention, or DWPE 
to CFSAN/Office of Compliance/Division of Enforcement (HFS-605):  
1.  For all dairy products: 
  7 Contains Nonbinding Recommendations 
  EHEC O157:H7 or other enterohemorrhagic Escherichia coli, or vegetative cells 
of Clostridium botulinum, or Clostridium botulinum toxin found in one or more 
subsamples; or 
  Escherichia coli found at levels greater than 10 MPN per gram in two or more 
subsamples or greater than 100 MPN per gram in one or more subsamples; or 
  Staphylococcus aureus found at levels greater than or equal to 104 cfu/g in one or 
more subsamples; or 
  Bacillus cereus found at levels greater than or equal to 104 cfu/g in one or more 
subsamples. 
2.  For cheese and related cheese products made from pasteurized bovine milk: 
  Alkaline phosphatase level in: 
o  Brick, semisoft, and semisoft part-skim cheeses - greater than 5 g/0.25 g (20 
micrograms phenol equivalent per gram) in one or more subsamples;  
o  Limburger cheese - greater than 4 g/0.25g (16 micrograms phenol equivalent 
per gram) in one or more subsamples for Limburger cheese;  
o  All other cheese and related cheese products - greater than 3 g/0.25g (12 
micrograms phenol equivalent per gram) in one or more subsamples. 
3.  For all other dairy products made from bovine milk, except cheese and related cheese 
products: 
  Alkaline phosphatase level is greater than or equal to 2.0 micrograms phenol 
equivalents per gram in one or more subsamples. 
 
Other Considerations 
 
This guidance does not establish acceptable levels of pathogens, nontoxigenic 
Escherichia coli, or alkaline phosphatase in dairy products.  FDA may choose to take 
action against adulterated food that does not meet the criteria under this Section. 
 
V.  Specimen Charges: 
 
Domestic Seizure 
 
A. Pathogens 
Salmonella species, EHEC O157:H7 or other enterohemorrhagic Escherichia coli, 
Campylobacter jejuni, Yersinia enterocolitica, vegetative cells of Clostridium 
botulinum, Clostridium botulinim toxin, Staphylococcus enterotoxin, Bacillus cereus 
enterotoxin 
The article of food was adulterated when introduced into and while in interstate 
commerce and is adulterated while held for sale after shipment in interstate commerce, 
within the meaning of the Act, 21 U.S.C. 342(a)(1), in that it bears and contains a 
poisonous or deleterious substance, namely ________,  which may render it injurious to 
health.  
 
  8 Contains Nonbinding Recommendations 
Staphylococcus aureus 
The article of food was adulterated when introduced into and while in interstate 
commerce and is adulterated while held for sale after shipment in interstate commerce, 
within the meaning of the Act, 21 U.S.C. 342(a)(4), in that it has been prepared, packed, 
and held under insanitary conditions whereby it may have been rendered injurious to 
health. 
 
Bacillus cereus 
The article of food was adulterated when introduced into and while in interstate 
commerce and is adulterated while held for sale after shipment in interstate commerce, 
within the meaning of the Act, 21 U.S.C. 342(a)(4), in that it has been prepared, packed, 
and held under insanitary conditions whereby it may have been rendered injurious to 
health.  
 
B. Nontoxigenic Escherichia coli 
The article of food was adulterated when introduced into and while in interstate 
commerce and is adulterated while held for sale after shipment in interstate commerce, 
within the meaning of the Act, 21 U.S.C. 342(a)(4), in that it has been prepared, packed, 
and held under insanitary conditions whereby it may have become contaminated with 
filth. 
 
C. Alkaline phosphatase 
The article of food was adulterated when introduced into and while in interstate 
commerce and is adulterated while held for sale after shipment in interstate commerce, 
within the meaning of the Act, 21 U.S.C. 342(a)(4), in that it has been prepared, packed, 
and held under insanitary conditions whereby it may have been rendered injurious to 
health.  
 
Import Detention 
 
A. Pathogens 
Salmonella species, EHEC O157:H7 or other enterohemorrhagic Escherichia coli, 
Campylobacter jejuni, Yersinia enterocolitica, vegetative cells of Clostridium 
botulinum, Clostridium botulinum toxin, Staphylococcus enterotoxin, Bacillus cereus 
enterotoxin 
The article of food is subject to refusal of admission pursuant to section 801(a)(3) of the 
FD&C Act in that it appears to be adulterated within the meaning of section 402(a)(1) of 
the FD&C Act in that it bears and contains a poisonous and deleterious substance, 
namely                           ,  which may render it injurious to health.  
 
Staphylococcus aureus 
The article of food is subject to refusal of admission pursuant to section 801(a)(3) of the 
FD&C Act  in that it appears to be adulterated within the meaning of section 402(a)(4) of 
the FD&C Act in that it has been prepared, packed, and held under insanitary conditions 
whereby it may have been rendered injurious to health. 
 
  9 Contains Nonbinding Recommendations 
Bacillus cereus 
The article of food is subject to refusal of admission pursuant to section 801(a)(3) of the 
FD&C Act  in that it appears to be adulterated within the meaning of section 402(a)(4) of 
the FD&C Act in that it has been prepared, packed, and held under insanitary conditions 
whereby it may have been rendered injurious to health. 
 
B. Nontoxigenic Escherichia coli 
The article of food is subject to refusal of admission pursuant to section 801(a)(3) of the 
FD&C Act in that it appears to be adulterated within the meaning of section 402(a)(4) of 
the FD&C Act in that it has been prepared, packed, and held under insanitary conditions 
whereby it may have become contaminated with filth. 
 
C. Alkaline phosphatase 
The article of food is subject to refusal of admission pursuant to section 801(a)(3) of the 
FD&C Act  in that it appears to be adulterated within the meaning of section 402(a)(4) of 
the FD&C Act in that it has been prepared, packed, and held under insanitary conditions 
whereby it may have been rendered injurious to health.* 
   
*Material between asterisks is new or revised.*  
 
Issued: 10/1/80  
Revised: 7/1/83, 8/1/86, 3/95, 8/96, 12/2010 
  10