Guidance for Industry: Standards for the 
Growing, Harvesting, Packing, and 
Holding of Sprouts for Human 
Consumption 
 
You may submit electronic or written comments regarding this guidance at any time. Submit 
electronic comments to https://www.regulations.gov. Submit written comments to the Dockets 
Management Staff (HFA-305), Food and Drug Administration, 5630 Fishers Lane, rm. 1061, 
Rockville, MD 20852. All comments should be identified with Docket No. FDA-2017-D-0175.  
For questions regarding this document contact FDAs Technical Assistance Network by 
submitting your question at https://www.fda.gov/food/food-safety-modernization-act-fsma/fsma-
technical-assistance-network-tan. 
 
U.S. Department of Health and Human Services 
Food and Drug Administration 
Center for Food Safety and Applied Nutrition  
 
September 2023 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
OMB Control No. 0910-0816 
Current expiration date available at https://www.reginfo.gov  
See additional PRA statement in Section IX of this guidance 
 
    Contains Nonbinding Recommendations 
Table of Contents 
 
I. Introduction ............................................................................................................................... 4 
II. Background .............................................................................................................................. 6 
A. Other FDA Efforts Related to Sprouts ................................................................................... 6 
B. Coverage of the Produce Safety Rule ..................................................................................... 7 
C. Coverage of Subpart M .......................................................................................................... 8 
D. Compliance Dates for Sprouts .............................................................................................. 11 
E. General Sprout Production .................................................................................................... 11 
III. Cleaning and Sanitizing ....................................................................................................... 12 
A. Frequency of Cleaning and Sanitizing ................................................................................. 12 
B. Cleaning and Sanitizing Policies and Recordkeeping .......................................................... 15 
C. Cleaning ................................................................................................................................ 16 
D. Sanitizing .............................................................................................................................. 17 
E. Verification of Cleaning and Sanitizing ............................................................................... 18 
F. Cleaning and Sanitizing Conducted in Response to Suspected or Known  Contamination . 18 
IV. Agricultural Water in Sprout Operations........................................................................... 19 
A. Numerical Microbial Quality Criterion for Agricultural Water Used in a  Sprouting 
Operation ( 112.44(a)) ............................................................................................................. 20 
B. Agricultural Water Source Type and Treatment Status ........................................................ 21 
C. Safe and of Adequate Sanitary Quality for Its Intended Use ( 112.41) .............................. 23 
D. Reuse of Sprout Irrigation Water ......................................................................................... 24 
E. Agricultural Water Testing Frequency, Sampling, and Test Methods ................................. 25 
F. Post-Harvest Water Management ......................................................................................... 26 
V. Seeds for Sprouting ................................................................................................................ 27 
A. Seed Sourcing, Receiving, Handling and Storage ................................................................ 28 
1. Seed Sourcing .................................................................................................................... 28 
2. Seed receiving .................................................................................................................... 29 
3. Visual inspection of seeds and their packaging ................................................................. 30 
4. Seed testing ........................................................................................................................ 31 
5. Seed storage ....................................................................................................................... 32 
B. Seed Treatment ..................................................................................................................... 32 
1. Choosing a seed treatment ................................................................................................ 34 
2. Seed treatment efficacy ...................................................................................................... 35 
3. Using pre-treated seeds ..................................................................................................... 38 
4. Additional considerations for treating seeds for sprouting ............................................... 38 
C. Corrective Actions for Seeds That May Be Contaminated with a Pathogen........................ 40 
VI. Environmental Monitoring .................................................................................................. 42 
A. Principles for Developing an Environmental Monitoring Plan ............................................ 43 
B. The Written Environmental Monitoring Plan ....................................................................... 44 
C. Developing a Sampling Plan ................................................................................................ 45 
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  Contains Nonbinding Recommendations 
D. Testing for Listeria spp. or L. monocytogenes ..................................................................... 46 
E. Person(s) Collecting Samples ............................................................................................... 46 
F. Establishing Sample Collection Locations and Frequency ................................................... 47 
1. Identifying sample collection locations ............................................................................. 47 
2. Deciding on the number of food contact surface and non-food contact surface sampling 
sites ........................................................................................................................................ 47 
3. Identifying sampling frequency ......................................................................................... 48 
G. Timing of Sample Collection ............................................................................................... 50 
H. Sample Collection and Shipping .......................................................................................... 51 
I. Choosing a Laboratory ........................................................................................................... 51 
J. Choosing a Test Method for Listeria spp. or L. monocytogenes ........................................... 52 
K. Documenting and Interpreting Test Results ......................................................................... 54 
1. Sampling plan that specifies testing for Listeria spp. (recommended approach) ............. 54 
2. Sampling plan that includes testing for L. monocytogenes ............................................... 55 
L. Developing a Corrective Action Plan and Taking Corrective Actions ................................. 56 
1. Corrective action plan ....................................................................................................... 56 
2. Implementing corrective actions ....................................................................................... 56 
3. Corrective actions if you detect Listeria spp. on a food contact surface .......................... 57 
4. Corrective actions if you detect Listeria monocytogenes on a food contact surface or non-
food contact surface............................................................................................................... 63 
5. Corrective actions if you detect Listeria spp. on a non-food contact surface ................... 64 
M. Analysis of Data for Trends ................................................................................................. 70 
VIII. Recordkeeping ................................................................................................................... 70 
A. Recordkeeping Overview ..................................................................................................... 71 
1. General requirements ........................................................................................................ 71 
2. Duplication not required ................................................................................................... 74 
3. Record retention and availability ...................................................................................... 74 
4. Format ............................................................................................................................... 74 
B. Supervisory Review of Records ........................................................................................... 75 
C. Sprouts-Specific Record Requirements and Recommendations .......................................... 76 
1. Enforcement Discretion for Requirements of Subpart M (applies to soil or substrate 
grown sprout types that are harvested by the customer before use) ..................................... 76 
2. Records for Seeds for Sprouting ........................................................................................ 76 
3. Required Records for Spent Sprout Irrigation Water ........................................................ 79 
VIII. Appendices ......................................................................................................................... 85 
Appendix 1. Aseptic Sampling.................................................................................................. 85 
Appendix 2. Collection Procedures  Environmental Sampling ............................................... 87 
Appendix 3. Potential Sources of L. monocytogenes for Sampling in a Sprout Operation ...... 91 
IX. Paperwork Reduction Act of 1995 ...................................................................................... 94 
X. References ............................................................................................................................... 95 
 
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  Contains Nonbinding Recommendations 
Guidance for Industry: Standards for the 
Growing, Harvesting, Packing, and 
Holding of Sprouts for Human 
Consumption 
 
This guidance represents the current thinking of the Food and Drug Administrations (FDA or 
we) on this topic. It does not establish any rights for any person and is not binding on FDA or the 
public. You can use an alternative approach if it satisfies the requirements of the applicable 
statutes and regulations. To discuss an alternative approach, contact FDAs Technical Assistance 
Network by submitting your question at https://www.fda.gov/food/food-safety-modernization-
act-fsma/fsma-technical-assistance-network-tan.  
I. Introduction 
This guidance is intended for those persons (you) who grow, harvest, pack and/or hold sprouts 
covered by Subpart M of our final rule, published in the Federal Register (80 FR 74353) on 
November 27, 2015, entitled, Standards for the Growing, Harvesting, Packing, and Holding of 
Produce for Human Consumption (the Produce Safety Rule or the Rule). 
The Produce Safety Rule established for the first time U.S. Federal requirements for the growing, 
harvesting, packing, and holding of produce for human consumption, including sprouts (Title 21 
Code of Federal Regulations part 112 (21 CFR part 112)). The Rule focuses on certain 
conditions and practices identified as common routes of contamination of produce (similar to the 
areas covered by the 1998 Guidance, Guide to Minimize Microbial Food Safety Hazards for 
Fresh Fruits and Vegetables (GAPs Guide)) (Ref. 1). The Rule establishes requirements 
addressing common routes of microbial contamination, including: agricultural water; biological 
soil amendments of animal origin; worker health and hygiene; equipment, tools, buildings and 
sanitation; and domesticated and wild animals.  
Sprouts represent a distinct food safety concern because the conditions under which sprouts are 
produced (i.e., temperature, water activity, pH and available nutrients) are also ideal for the 
growth of pathogens, if present (Ref. 2). Between 1996 and 2020 in the United States, FDA 
observed 52 reported outbreaks of foodborne illness associated with sprouts. Together, it is 
estimated that these outbreaks resulted in at least 2700 cases of illness, 200 hospitalizations, and 
three deaths (Ref. 3, Ref. 4, Ref. 5, Ref. 6, Ref. 7, and Ref. 8). During this timeframe, sprouts 
have been associated with outbreaks of several different pathogens, including Salmonella spp., 
Listeria monocytogenes, E. coli O157:H7, and several types of non-O157:H7 pathogenic E. coli 
(i.e., E. coli O157:NM (H-), E. coli O104:H4, E. coli O26, E. coli O121, E. coli O103) (Ref. 9 and 
Ref. 10). In foodborne illness outbreaks associated with sprouts where the source of 
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  Contains Nonbinding Recommendations 
contamination was identified, epidemiological investigations often identify the most likely 
source of contamination as seeds used for sprouting (Ref. 2 and Ref. 11). However, poor 
sanitation and unhygienic practices at the sprout operation have also contributed to the 
contamination of sprouts (Ref. 2, Ref. 6, Ref. 12, Ref. 13, and Ref. 14). 
Because the distinctive practices and conditions for growing sprouts present unique risks, we 
established sprout-specific requirements in Subpart M (Sprouts) of the Produce Safety Rule. 
Subpart M of the Rule builds on sprout production practices similar to areas covered in our 1999 
Sprout Guidances (discussed further below, now withdrawn). Sprout operations subject to the 
Produce Safety Rule must comply with all applicable requirements in the Rule, including, but not 
limited to, all applicable requirements in Subpart M. 
The requirements in the Produce Safety Rule are directed specifically to covered farms (as that 
term is defined in the Rule) that grow, harvest, pack or hold covered produce, including sprouts.  
Covered farms that grow, harvest, pack or hold sprouts are referred to in this guidance as sprout 
operations, or you. Produce that is covered by the Rule is referred to as covered produce.  
Neither the Produce Safety Rule nor this guidance document is directed to growing, 
conditioning, or distributing seed for sprouting or to the handling of sprouts at a retail food 
establishment. However, as noted in our prior Sprout Guidances and our May 2009 letter to 
suppliers and distributors of seed for sprouting and sprout operations (Ref. 15 and Ref. 16), 
everyone in the sprout supply chain has a responsibility to help ensure food safety. FDA has also 
issued the final guidance document regarding good agricultural practices for seed for sprouting, 
entitled Reducing Microbial Food Safety Hazards in the Production of Seed for Sprouting: 
Guidance for Industry (Ref. 17). The Produce Safety Rule does not address chemical or 
physical hazards. However, you have a responsibility to ensure that your sprouts are not 
adulterated or misbranded under the Federal Food, Drug, and Cosmetic Act (FD&C Act) (21 
U.S.C.  301 et seq.) and are produced in compliance with all applicable laws and regulations. 
Under section 402(a)(1) of the FD&C Act, a food is adulterated if it bears or contains any added 
poisonous or deleterious substance which may render it injurious to health, and such substances 
may include or otherwise result from physical and chemical (including radiological) 
contamination. 
This guidance provides our current thinking and recommendations to assist sprout operations 
subject to the Produce Safety Rule primarily in complying with the sprout-specific requirements 
in Subpart M. This guidance also briefly discusses certain requirements (in Subparts E and O of 
the Rule relating to Agricultural Water and Records) of particular relevance to a sprout 
operation. Some requirements of Subpart M and other subparts to which sprouts are subject (e.g., 
Subpart M, 112.147, related to spent sprout irrigation water; Subpart C, related to training; and 
Subpart L, related to equipment, tools, and buildings) are covered in the draft reissued sections 
(Ref. 18). In addition, this guidance may also be useful to sprout operations that are not subject 
to the Produce Safety Rule that voluntarily choose to follow the standards established in the 
Rule. In the development of this guidance, we particularly considered industry and international 
documents related to food safety and hygienic production of sprouts (Ref. 19, Ref. 20, and Ref. 
21). We have incorporated aspects of these documents that are consistent with our laws, 
regulations, and existing policies. 
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  Contains Nonbinding Recommendations 
Some of the material in this guidance relates to regulatory requirements of the Produce Safety 
Rule that are also covered in the draft Standards for the Growing, Harvesting, Packing, and 
Holding of Produce for Human Consumption: Guidance for Industry (general produce 
compliance and implementation draft guidance (83 FR 53196) issued in October 2018) (Ref. 22). 
At the present time, the material on these overlapping topics (e.g., records, cleaning and 
sanitizing) is consistent between the two guidance documents. This final guidance focuses 
specifically on insights drawn from FDAs experience with sprout operations, such as from 
inspections and sprout-associated foodborne illness outbreak investigations and elaborates on 
how the broader records and cleaning and sanitizing standards in part 112 could apply to and be 
implemented in a sprout operation.  
In general, FDAs guidance documents do not establish legally enforceable responsibilities. 
Instead, guidances describe the Agencys 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 
A. Other FDA Efforts Related to Sprouts 
On October 27, 1999, we published a notice of availability in the Federal Register (64 FR 
57893) for two guidance documents to inform all parties involved in the production of sprouts 
(i.e., producers, conditioners, and distributors of seeds used for sprouting, and sprout operations) 
that sprouts had been recognized as an important cause of foodborne illness and to provide 
recommendations for preventive controls that we believed should be taken immediately to reduce 
the likelihood of sprouts serving as a vehicle for foodborne illness. We refer to these prior (now 
withdrawn) guidance documents collectively as the 1999 Sprout Guidances. 
FDA and our food safety partners in the public and private sectors have engaged in education 
and outreach to the sprout industry to promote adoption of our recommendations. We have also 
worked with the sprout industry to advance the scientific knowledge applicable to enhancing the 
safety of sprouts. For example, in 2000, we collaborated with the California Department of 
Public Health, in cooperation with the sprout industry and academia, to develop an educational 
video series entitled, Safer Processing of Sprouts (Ref. 23). In addition, we have provided 
technical assistance to the Illinois Institute of Technologys Institute for Food Safety and Health 
(IIT IFSH) Sprout Safety Taskforce in  developing their US Sprout Industry Production Best 
Practices (Ref. 20). 
We also have been working with the Sprout Safety Alliance (SSA), a public-private partnership, 
since 2012 to develop a standardized curriculum and training and outreach programs for 
stakeholders in the sprout industry to enhance the industrys understanding and implementation 
of the requirements of the Produce Safety Rule, and of best practices for improving sprout safety 
(Ref. 24, Ref. 25, and Ref. 26). The SSA is composed of representatives from the sprout industry, 
retailers, academia, and federal, state, and local food safety agencies. The SSA is funded by a 
grant from FDA to IIT IFSH. 
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  Contains Nonbinding Recommendations 
B. Coverage of the Produce Safety Rule 
Under  112.1 of the Produce Safety Rule, unless specifically excluded under  112.2, food that 
is produce (as that term is defined in the Rule), and that is a raw agricultural commodity (RAC), 
is covered by the Rule. This includes a produce RAC that is grown domestically and a produce 
RAC that will be imported or offered for import into any State or territory of the United States, 
the District of Columbia, or the Commonwealth of Puerto Rico. Covered farms (i.e., those 
subject to the Produce Safety Rule) must comply with all applicable requirements of the Rule 
when conducting a covered activity on covered produce (see  112.4). There are certain 
exclusions, exemptions and limitations on which farms are covered farms (see  112.4). For 
example, sprout operations for which, on a rolling basis, the average annual monetary value of 
produce sold during the previous 3-year period, adjusted for inflation, is less than or equal to 
$25,000, are not covered by this regulation ( 112.4(a)). See https://www.fda.gov/food/food-
safety-modernization-act-fsma/fsma-inflation-adjusted-cut-offs for information on FDA Food 
Safety Modernization Act (FSMA) inflation adjusted cut-offs. See also Standards for Produce 
Safety, Coverage and Exemptions/Exclusions for 21 Part 112, available at 
https://www.fda.gov/media/94332/download to assist you in determining which provisions of the 
Produce Safety Rule, if any, apply to you. 
Sprouts are produce as we defined that term in the Produce Safety Rule (see  112.3). Sprouts are 
also RACs when they are in their raw or natural state (see  112.3 and section 201(r) of the 
FD&C Act, defining raw agricultural commodity as any food in its raw or natural state, 
including all fruits that are washed, colored, or otherwise treated in their unpeeled natural form 
prior to marketing.). Therefore, sprouts in their raw or natural state are covered produce, except 
as otherwise provided in  112.2. 
If sprouts are made into processed food(s), those processed foods are not covered by the Produce 
Safety Rule (see  112.2(a)(3)). An example of a processed food made using sprouts is sprouted 
seed butter. This does not mean that sprout RACs that will be made into processed food are 
themselves exempt from the Produce Safety Rule simply because those RACs will later be 
transformed into processed food. Rather, it means that the Produce Safety Rule only applies 
during the time that the sprouts are RACs. Other requirements, such as those in 21 CFR Part 117, 
may apply to any manufacturing/processing of the sprout RACs into processed food, depending 
on the circumstances. 
Sprouts do not qualify for the exemption from the Produce Safety Rule in  112.2(a)(1) for 
produce that is rarely consumed raw. This provision contains a list of produce commodities (such 
as potatoes) for which we have analyzed dietary consumption patterns and determined such 
commodities are rarely consumed raw. This exemption applies only to the commodities 
identified in  112.2(a)(1), none of which are sprouts. 
Some sprouts may be eligible for exemption from the Produce Safety Rule under  112.2(b), if 
they will receive commercial processing that adequately reduces the presence of microorganisms 
of public health significance. Examples of commercial processing that adequately reduces the 
presence of microorganisms of public health significance appear in  112.2(b)(1). This 
exemption requires covered farms to establish and maintain certain documentation and 
disclosures set forth in  112.2(b)(2)-(3): 1) disclosures to customers; and 2) annual written 
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  Contains Nonbinding Recommendations 
assurances obtained from customers. Sprouts that receive commercial processing (e.g., canned, 
shelf-stable mung bean sprouts) could potentially qualify for this exemption from the Produce 
Safety Rule, but only if the commercial processing adequately reduces the presence of 
microorganisms of public health significance and all documentation and disclosure requirements 
are met.1 We note that simply drying/dehydrating sprouts may not adequately reduce the 
presence of microorganisms of public health significance (Ref. 27 and Ref. 28) and therefore 
would not make them eligible to receive the commercial processing exemption from the Rule. 
Some sprout operations may be eligible for a qualified exemption and associated modified 
requirements in a calendar year if, during the previous 3-year period preceding the applicable 
calendar year, the average annual monetary value of the food (as defined in  112.3) the sprout 
operation sold directly to qualified end-users (as defined in  112.3) during such period exceeded 
the average annual monetary value of the food the sprout operation sold to all other buyers 
during that period; and the average annual monetary value of all food (as defined in  112.3) the 
sprout operation sold during the 3-year period preceding the applicable calendar year was less 
than $500,000, adjusted for inflation. (FDA makes available the updated inflation-adjusted 
figures at https://www.fda.gov/food/food-safety-modernization-act-fsma/fsma-inflation-adjusted-
cut-offs.)  Section 112.6(b) describes the modified requirements applicable to a qualified exempt 
operation (e.g., labeling). Section 112.7 requires the sprout operation eligible for the qualified 
exemption, provided for in  112.5, to establish and keep adequate records necessary to 
demonstrate that the sprout operation satisfies the criteria for a qualified exemption (e.g., dated 
sales receipts), including a written record reflecting that the owner, operator, or agent in charge 
of the sprout operation has performed an annual review and verification of the sprout operations 
continued eligibility for the qualified exemption. 
C. Coverage of Subpart M 
The requirements in 21 CFR part 112, Subpart M apply to the growing, harvesting, packing and 
holding of all sprouts except sprouts that are grown in soil or non-soil substrates (e.g., mats, 
perlite or other growth media) and that are harvested above the soil or substrate line without their 
roots (see  112.141). We determined that soil- or substrate-grown sprouts that are harvested 
above the soil or substrate line, such that their roots are not harvested for human consumption, 
do not present the same risks as other types of sprouts and, therefore, we excluded them from the 
 
1 In the Federal Register of January 5, 2018 (83 FR 598), we published a notification of availability of a guidance 
document titled Policy Regarding Certain Entities Subject to the Good Manufacturing Practice and Preventive 
Controls, Produce Safety, and/or Foreign Supplier Verification Programs. In that guidance document, we stated 
that we intend to exercise enforcement discretion regarding certain written assurance requirements, including those 
in 21 CFR part 112 (the Produce Safety Rule). We intend to exercise such discretion until we can complete a 
rulemaking process to consider options for the assurance requirements. https://www.fda.gov/regulatory-
information/search-fda-guidance-documents/guidance-industry-policy-regarding-certain-entities-subject-current-
good-manufacturing-practice-and. See also the Federal Register of March 14, 2022 (87 FR 14169), in which we 
published a notification of availability of a guidance document titled Current Good Manufacturing Practice and 
Preventive Controls, Foreign Supplier Verification Programs, Intentional Adulteration, and Produce Safety 
Regulations: Enforcement Policy Regarding Certain Provisions.  In that guidance document, we restated our intent 
to exercise enforcement discretion with respect to the Produce Safety Rule assurance requirements, as previously 
indicated in the 2018 guidance document described immediately above. https://www.fda.gov/regulatory-
information/search-fda-guidance-documents/guidance-industry-current-good-manufacturing-practice-and-
preventive-controls-foreign-supplier.  
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sprout-specific requirements in Subpart M (80 FR 74353 at 74497). However, the requirements 
of Subpart M do apply to soil- or substrate-grown sprouts that are harvested with the roots. If you 
use soil or substrate as a growth medium in your operation, in addition to all other relevant 
requirements of the Produce Safety Rule, you must comply with the applicable requirements in 
21 CFR part 112, Subpart F (Biological Soil Amendments of Animal Origin and Human Waste).  
We discuss the requirements of Subpart F in the draft compliance and implementation guidance 
(Ref. 22).  
We recognize that sprout operations may sell certain soil- or substrate-grown sprout types, such 
as wheatgrass, in a tray used for growing, with the soil/substrate and root intact, to commercial 
entities, including facilities that are required to register with FDA under section 415 of the 
FD&C Act, restaurants, retail food establishments, and non-profit food establishments. In such 
cases, the operation that has harvested the sprouts with the roots and these sprouts are ordinarily 
subject to Subpart M (see  112.141). However, we understand that, in some cases, such sprouts 
may then be cut above the soil and/or substrate line at the commercial entity immediately before 
use or sale to the consumer. When a  sprout operation sells sprouts with the roots intact in soil or 
substrate, and the commercial entity that receives them will cut the  sprouts above the soil or 
substrate line before use, we intend to exercise enforcement discretion for the requirements of 
Subpart M if the sprout operation annually collects written assurances from the commercial 
entity stating that the sprouts will be cut above the soil or substrate line before use (see also 
Section VII.C.1).  
Note that soil- or substrate-grown sprouts harvested above the soil line are still considered 
covered produce and, unless exempt or excluded under the provisions of 21 CFR part 112, 
Subpart A, are subject to all other applicable requirements of the Produce Safety Rule. The 
standards in Subpart M may be useful to sprout operations that produce soil- or substrate-grown 
sprouts that are harvested above the soil or substrate line that voluntarily choose to follow these 
standards, in addition to complying with the required provisions of all other subparts in the 
Produce Safety Rule. 
We are aware that certain sprouts are grown hydroponically in trays, and then harvested without 
the root at either the sprout operation or by the customer. As we previously stated in the 
preamble to the final rule (80 FR 74353 at 74497, comment/response 364), all hydroponically 
grown sprouts are subject to the requirements of Subpart M. Under typical conditions for 
growing hydroponic sprouts, water circulates between sprouts in the same growing unit, such 
that any pathogens present in the seed or introduced during sprouting can spread throughout the 
production lot of sprouts (Ref. 2, Ref. 29, and Ref. 30).  
We note that microgreens and sprouts are different products. This interpretation is consistent 
with our prior Sprout Guidances and with other public and private standards (e.g., IIT IFSH 
Sprout Taskforce US Sprout Industry Production Best Practices document (Ref. 20), Food Safety 
Australia New Zealand (FSANZ) standards for sprouts (Ref. 31)). Historically, the primary 
criterion we have used to distinguish between the two product categories has been the growth 
stage of the leaves. Sprouts are usually harvested when the cotyledons (i.e., seed leaves) are still 
un- or under-developed and true leaves have not begun to emerge. In contrast, microgreens reach 
a later stage of growth, typically associated with the emergence of true leaves. Unlike sprouts 
covered by Subpart M, microgreens are typically grown in soil or substrate and harvested above 
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the soil or substrate line. Because microgreens are not sprouts, they are not subject to the 
requirements in Subpart M (Ref. 32). However, microgreens are considered covered produce for 
the purposes of the Produce Safety Rule and, unless exempt or excluded under the provisions in 
Subpart A, microgreens are subject to the other subparts of the  Produce Safety Rule (80 FR 
74353 at 74497, comment/response 363). 
The following figure provides factors to consider when determining whether sprouts are subject 
to the sprout-specific requirements of Subpart M of the Produce Safety Rule.  
Figure 1. Determining Coverage under Subpart M of the Produce Safety Rule 
 
 
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  Contains Nonbinding Recommendations 
Note that throughout this guidance, for the ease of the reader, we often refer collectively to 
everything, including beans, sprouted to produce sprouts for human consumption simply as 
seeds. In the Rule, we used the phrase seeds or beans to remove any potential confusion as to 
whether beans for sprouting were included. References to seeds in this guidance should be read 
to include things other than seeds that are sprouted to produce sprouts for human consumption, 
such as beans.  
In addition, the definition of produce includes sprouts irrespective of seed source (see  112.3). 
We consider sprouts to be a distinct category of produce that includes many different varieties 
(e.g., alfalfa, mung). Sprouts (i.e., alfalfa, mung and soybean sprouts) were evaluated to 
determine whether they are rarely consumed raw, and therefore exempt from the Produce Safety 
Rule (see 80 FR 74353 at 74392, comment/response 68). Sprouts did not meet the threshold 
criteria for the rarely consumed raw exemption and are therefore considered covered produce. 
The mature plant and/or the seeds used to grow sprouts may or may not fall under the definition 
of produce. For example, wheatgrass sprouts and soybean sprouts have long been considered 
sprouts by the food industry and FDA and are covered produce under the Produce Safety Rule. 
However, for both wheat and soybeans, the mature crops are considered food grains, which do 
not fall under the definition of produce and are not subject to the Produce Safety Rule. Sprouts 
may also be produced from seeds of produce that is considered to be rarely consumed raw, such 
as beet or pumpkin sprouts. While the mature produce may be rarely consumed raw, and 
therefore exempt from the Produce Safety Rule, the sprouts produced from the seeds of these 
crops are still covered produce under the Produce Safety Rule because we consider sprouts to be 
a distinct commodity from the mature form, and sprouts have not met the threshold criteria for 
the rarely consumed raw exemption.  
D. Compliance Dates for Sprouts 
Sprout operations covered by the Produce Safety Rule and subject to Subpart M must be in 
compliance with all applicable provisions of the Produce Safety Rule at this time. The extended 
compliance dates provided for covered farms producing other produce commodities to comply 
with the agricultural water requirements of Subpart E do not apply to the production of sprouts 
subject to Subpart M. Sprout operations eligible for the qualified exemption from the Produce 
Safety Rule are also expected to be in compliance with the applicable modified requirements at 
this time.  
The same compliance dates that apply to all other covered produce apply to sprouts that are not 
subject to Subpart M but are otherwise subject to the Produce Safety Rule (i.e., soil- or substrate-
grown sprouts harvested without their roots). 
E. General Sprout Production 
This section discusses common production practices for growing, harvesting, packing, packaging 
and holding of sprouted seeds. Later sections of this guidance describe the requirements of the 
Produce Safety Rule and provide recommendations for sprouts operations in conducting the 
different steps in sprout production.  
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Typically, sprout production consists broadly of the steps depicted in Figure 2. Your operation 
may add to or omit some of these practices (or do them in a different order) depending on a 
number of factors, including: the type of seeds you sprout; whether the required seed treatment is 
applied by you, your seed supplier, or both; and the size and resources of your operation.  
Figure 2. Typical Sprout Production Processes (Adapted from Ref. 2) 
Seed Receipt  Seed Storage  Initial Seed Rinse  Seed Treatment  Pre-germination 
Seed Soak  Germination and Growth  Microbial Testing of Spent Sprout Irrigation Water 
(or in-process sprouts)  Harvest Wash/Drain Sprouts Bulk Cool/Spin Dry  Pack 
and/or Package  Cooling & Storage Distribution 
III. Cleaning and Sanitizing 
Cleaning and sanitizing are different, and important, steps that are critical to the safety of your 
finished sprouts. Cleaning refers to the practice of removing organic material and other debris 
from surfaces. The Produce Safety Rule defines sanitize to mean to adequately treat cleaned 
surfaces by a process that is effective in destroying vegetative cells of microorganisms of public 
health significance, and in substantially reducing numbers of other undesirable microorganisms, 
but without adversely affecting the product or its safety for the consumer ( 112.3). When 
cleaning and sanitizing is required, you must properly clean surfaces prior to sanitizing because 
many sanitizers will not be effective unless food and dirt have been removed from the surface 
first (see definition of sanitize in  112.3, to adequately treat cleaned surfaces ... (emphasis 
added)). Sanitizing surfaces is usually achieved through chemical means. 
This section discusses cleaning and sanitizing policies and procedures, frequencies, verification 
activities, recordkeeping, and corrective actions to take in response to suspected or known 
contamination. 
A. Frequency of Cleaning and Sanitizing 
Section 112.123(d)(1) is a general requirement that applies to all covered farms, and requires you 
to inspect, maintain, clean and, when necessary and appropriate, sanitize all food contact 
surfaces (FCSs) of equipment and  tools used in covered activities as frequently as reasonably 
necessary to protect against contamination of covered produce. For operations growing sprouts 
covered by Subpart M, we determined that it is necessary and appropriate to sanitize all such 
FCSs used to grow, harvest, pack, or hold sprouts after cleaning the FCSs and therefore we 
specifically require both cleaning and sanitizing for such FCSs prior to contact with sprouts or 
seeds used to grow sprouts, as reflected in  112.143(b).  
Food contact surfaces means those surfaces that contact human food and those  surfaces from 
which drainage, or other transfer, onto the food or onto surfaces that contact the food ordinarily 
occurs during the normal course of operations. Food contact surfaces includes food contact 
surfaces of equipment and tools used during harvest, packing, and holding ( 112.3). Food 
contact surfaces also includes FCSs of equipment and tools used in growing sprouts, such as 
those that contact seeds for sprouting (see e.g.,  112.143(b)). In a sprouting operation, FCSs 
include, for example: trays or drums used for sprouting; interior surfaces of  containers used for 
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seed rinsing, seed treatment, and pre-germination seed soaking; and counters that come into 
contact with sprouts during packing and/or packaging. 
While the appropriate frequency of cleaning and sanitizing may vary based on your specific 
production practices, in general, we recommend cleaning and sanitizing of FCSs used in sprout 
operations at least daily to meet the requirements of  112.123(d)(1) and 112.143(b). For 
example, you should clean and sanitize all FCSs in the production environment at the end of each 
production day. We recommend that initial sanitizing, of any surface, occur following the 
completion of cleaning. You may determine that there are some situations where it is necessary 
to re-sanitize an FCS in the morning after cleaning and sanitizing it the night before. To illustrate 
the requirement in  112.143(b) that FCSs be cleaned and sanitized before contact with sprouts 
or seeds used to grow sprouts, we provide the following non-exhaustive list of examples of when 
you must incorporate a cleaning and sanitizing step for FCSs: 
  Before contact with a different production batch of sprouts (e.g., when more than one 
production batch of sprouts, including seeds for a different production batch of sprouts, 
will contact the same FCSs on the same day). Cleaning and sanitizing FCSs between 
production batches of sprouts minimizes the potential for cross-contamination from one 
production batch to the next, and also could reduce the extent of corrective actions 
needed in response to a positive pathogen (or indicator organism) test result (e.g., from a 
spent sprout irrigation water, sprouts, or environmental sample), because it could serve as 
a point at which the risk of contamination of subsequent production batches is 
substantially reduced (see Section V (Sampling and Testing of Spent Sprout Irrigation 
Water (or In-Process Sprouts)) in the draft re-issued sections (Ref. 18)) and Section VI 
(Environmental Monitoring) in this document);  
  Before the first time the FCS is used in your sprout operation to contact seeds for 
sprouting or sprouts. FCSs of such tools or equipment may have been contaminated 
during manufacturing, storage, or transport, or, if they were repurposed from another use, 
may have been contaminated during that use;  
  Before contact with seeds or sprouts after storage for an extended period (e.g., several 
days, weeks, or months) since the FCSs of the equipment and tools may have become 
contaminated during storage; 
  Before contact with seeds or sprouts after the FCS has been exposed to a source of 
contamination (e.g., contaminated water, human or animal fecal material, animal or insect 
activity); and 
  Before contact with seeds or sprouts after contacting food other than sprouts, regardless 
of whether the food is produce that is covered by the Produce Safety Rule, because of the 
risk that pathogens transferred from these other foods may multiply in the unique 
conditions in which sprouts are grown. 
Surfaces that are used continuously for more than a day, such as bins or drums that contain a 
single production batch of sprouts during germination and growth periods lasting more than a 
day, do not need to be cleaned or sanitized on a daily basis while they continuously contain or 
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otherwise contact only one production batch of  growing sprouts. We consider such uses to be a 
single, continuous instance of contact. The end of one production batch and the beginning of the 
next should be considered a break in the single, continuous instance of contact, warranting 
cleaning and sanitizing of the surfaces between batches. 
Additionally, you must maintain and clean all non-FCSs of equipment and tools subject to 
Subpart L that are used during harvesting, packing, and holding as frequently as reasonably 
necessary to protect against contamination of sprouts ( 112.123(d)(2)). You may consider 
sanitizing your non-FCSs on the same schedule as your FCSs. 
While the rule does not require sanitizing of non-FCSs, we recommend that sprout operations 
sanitize even non-FCSs on the same schedule that we recommend for cleaning FCSs because of 
the particularly high-risk nature of sprout production. We recommend cleaning and sanitizing all 
non-FCSs in accordance with the frequencies in Table 1 below. The recommendations in the 
following table are adapted from a 1999 publication by Tompkin et al. (Ref. 33). If the results of 
your environmental monitoring, spent sprout irrigation water testing, or product testing indicate a 
food safety concern, you should consider increasing the frequency of cleaning and sanitizing as 
part of an overall corrective action plan. If the manufacturer of equipment used in your operation 
recommends cleaning on a more frequent basis, we recommend that you increase this frequency 
to match the recommendations of the manufacturer. 
Table 1. Recommended Frequency of Cleaning and Sanitizing Non-Food Contact Surfaces 
for Sprout Operations 
Surface, Area, or Equipment  a
Frequency of Cleaning and Sanitizing  
Drains and floors  Daily 
Waste containers  Daily 
Cleaning tools (e.g., mops, brushes)  Daily 
Surfaces that have a greater potential to become  Daily 
a source of L. monocytogenes contamination 
(e.g., surfaces likely to be touched by employees 
who touch product or FCSs during operations, or 
areas where there may be a build-up of moisture 
or product residue) 
Condensate drip pans  Monthly 
Motor housings, external surfaces of enclosed  Monthly 
processing systems 
b Semi-annually 
Overhead piping, ceilings and walls  
Refrigerators (e.g., coolers) containing exposed  Quarterly 
RTE food 
Interiors of Ice Makers  Semi-annually 
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a 
Since production environments vary, it may be appropriate to increase cleaning frequencies depending on the 
specific circumstances of the product area. 
b 
We recommend that that you clean and sanitize walls and ceilings in close proximity to a production line at the 
same time as the production equipment (e.g., daily). 
B. Cleaning and Sanitizing Policies and Recordkeeping 
You should create policies that describe cleaning and sanitation procedures. These include 
schedules, needed materials, tools, and chemicals for specific cleaning and sanitizing tasks. You 
may need several policies describing various tasks throughout the production environment. Each 
piece of equipment, as well as the production environment itself, should be considered when 
developing your policies. While not required by the Produce Safety Rule, we recommend 
establishing these policies to promote consistent applications of your intended cleaning and 
sanitizing procedures by different workers. The following details should be included in each 
policy and periodically reviewed by management: 
  For what piece(s) of equipment or area(s) of the production environment the policy was 
created; 
  Who is responsible for the cleaning and sanitizing tasks (see discussion on  112.22(a)(3) 
in Section III. A. 1 (Training and Supervision of Personnel who Conduct Cleaning and 
Sanitizing Activities) of the draft re-issued sections (Ref. 18)); 
  When and how often each task is to be completed; 
  What tools and chemicals (for both cleaning and sanitizing) are needed; 
  How the chemicals are to be prepared, as applicable; 
  How the chemicals are to be used (including what precautions need to be taken), as 
applicable; and 
  How to properly clean and sanitize the piece(s) of equipment or work area(s), including 
any equipment disassembly that is necessary. 
You must establish and keep records of the date and method of cleaning and sanitizing of 
equipment used during growing operations for sprouts and all covered harvesting, packing, or 
holding activities ( 112.140(b)). The records should be sufficient to demonstrate that your 
established cleaning procedures have been achieved (e.g., through the use of a cleaning and 
sanitizing checklist). Maintaining records of the performed activities not only helps you ensure 
activities are appropriately performed, but also serves as documentation to show auditors or 
inspectors, as necessary, that the activities were properly performed. These records should be 
maintained each time cleaning and sanitizing is performed, even if performed more than once per 
day. 
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C. Cleaning 
You should establish your cleaning procedures based on the characteristics of the soils and debris; 
the type, design and age of equipment or tools; surface materials (e.g., stainless steel, plastic, 
concrete); the general effectiveness of your cleaning agent(s); label instructions; the results of 
sanitation verification activities (see Section III. E (Verification of Cleaning and Sanitizing)); 
and environmental monitoring results (see Section VI (Environmental Monitoring)). 
Factors that influence the effectiveness of cleaning procedures include, but are not limited to, the 
following: 
  Characteristics of the organic matter, sprouts, and other debris that would impact its 
removal from the equipment; 
  The surface material (e.g., plastic, stainless steel, concrete, etc.) to be cleaned and its 
condition and construction. You should consider the surface material to be cleaned when 
selecting cleaning agents/detergents; 
  Contact time of the cleaning agent/detergent with the surface(s) being cleaned; and 
  The duration and force of physical scrubbing and the pressure of water or air used during 
the removal of the organic material. 
Mops, brushes and other equipment used for cleaning and sanitizing should be durable and 
replaced immediately if damaged, cracked, or worn, to prevent the colonization of those tools by 
pathogens that could lead to contamination of product or FCSs. These tools and equipment 
should also be stored appropriately to prevent them from becoming soiled, and they should be 
cleaned and sanitized as needed. 
The following steps are an example of a typical wet cleaning procedure (Ref. 34 and Ref. 35); 
some steps could be inapplicable to your operation or could be modified (Ref. 34 and Ref. 36): 
1.  Pre-Clean: personnel use tools such as brushes, brooms, scrapers, and collection pans to 
remove heavy residue, plant material or debris from the equipment or tools; 
2.  Initial Rinse: personnel rinse the equipment or tools with water to remove additional 
organic material and residue that was loosened during the pre-clean step; scrubbing could 
be necessary during this step; 
3.  Washing with Cleaning Agent and Mechanical Action: personnel apply a cleaning agent in 
one of several forms, such as by spray or foam; personnel should ensure full coverage of 
the relevant surface with the cleaning agent for the time and at the temperature specified 
by the cleaning agent manufacturer; small equipment or tools or components of these can 
be washed in sinks or tanks containing a solution of the cleaning agent; mechanical action 
(e.g., scrubbing) ordinarily should be applied, using tools such as brushes or scouring 
pads, in order to fully remove organic material and residues; 
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4.  Post-Rinse: personnel apply water to remove loosened soil and debris along with the 
cleaning agent; after the post-rinse the equipment and tools could be allowed to dry prior 
to further activities; and 
5.  Inspection: personnel check the equipment or tools for any remaining soil or residue. 
D. Sanitizing 
Sanitizing is generally not effective unless it is preceded by a cleaning step, because residual 
organic material can insulate pathogens from the action of the sanitizing treatment (Ref. 34, Ref. 
36, Ref. 37, Ref. 38, and Ref. 39).    
The following should be considered when developing your sanitizing procedures and when 
sanitizing your  equipment, tools, or production environment: 
  The surfaces should be cleaned as described above in Section III. C (Cleaning) before 
sanitizing; 
  Sanitizing approaches should take into account the type, duration, application method, 
temperature, and concentration of the sanitizing or antimicrobial treatment (Ref. 36 and 
Ref. 40); 
  Verification activities, described below, should be conducted to evaluate the effectiveness 
of  your cleaning and sanitizing activities; and 
  Chemical sanitizing agents must be used according to label directions, in accordance with 
the  Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA). 
The following should be considered when selecting sanitizing agents: 
  The material of the surface that is to be sanitized; 
  The contact time of the sanitizing or antimicrobial treatment with the surface being 
treated; 
  Characteristics of water (e.g., hardness, pH, temperature) used to prepare the sanitizing 
agent; 
  The compatibility of the sanitizing agent with the cleaning chemical(s); and 
  Appropriateness of the sanitizing agent for use on food or FCSs.  
Consulting with cleaning and/or sanitizing agent suppliers, academia, industry associations, 
scientific literature, and government agencies may help you select the products that are  best 
suited for your operation. 
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E. Verification of Cleaning and Sanitizing 
You should verify the chemical concentration of the sanitizer(s) that you use each time you use 
them, with an appropriate test kit or analytical instrument. If analytical instruments are used to 
measure sanitizer concentrations, they must be accurate and  precise as necessary and 
appropriate in keeping with their purpose ( 112.124(a)), adequately  maintained ( 112.124(b)), 
and adequate in number for their designated use ( 112.124(c)). 
To verify the effectiveness of your cleaning and sanitizing procedures, we recommend that you 
use at least  one, or more, of the available test methods. Some of these tests, which include 
bioluminescence, adenosine triphosphate (ATP), and protein-based technologies, provide rapid 
results and allow for follow-up or intensified cleaning and sanitizing activities if results are 
above your established thresholds. However, it is critical to recognize that none of these tests is 
an appropriate substitute for environmental monitoring of Listeria spp. or L.  monocytogenes, as 
required for sprout operations by  112.145 (see Section VI (Environmental  Monitoring)). 
F. Cleaning and Sanitizing Conducted in Response to Suspected or Known 
Contamination 
In addition to the cleaning and sanitizing required by  112.123(d) and 112.143(b), you must 
clean and sanitize FCSs and the production environment as a corrective action measure if: 
  Sprouts or spent sprout irrigation water test positive for the pathogens E. coli O157:H7, 
Salmonella, or any pathogens meeting the criteria in  112.144(c). In such a situation, you 
must clean and sanitize the affected surfaces and surrounding areas ( 112.148(c)); or 
  Listeria spp. or L. monocytogenes is detected in the growing, harvesting, packing, or 
holding environment. In such a situation, you must clean and sanitize the affected 
surfaces and  surrounding areas ( 112.146(b)). 
Further, you should clean and sanitize FCSs and the production environment as a corrective 
action measure when: 
  You have used a lot of seeds that you have been subsequently notified has tested positive 
for a pathogen or has been implicated in a foodborne illness outbreak; 
  Your sprout operation is involved in a foodborne illness outbreak; or 
  You become aware of an occurrence of known or suspected contamination of your 
sprouts or sprout operation.  
In such situations you should consider the following  actions as a means of intensified cleaning 
and sanitizing: 
  Dismantling equipment further than is described in your cleaning and sanitizing 
procedures or is your normal practice; 
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  Intensified (e.g., more vigorous, longer duration) scrubbing of surfaces; 
  Identifying, cleaning, and sanitizing possible harborage sites for pathogens and possible 
cross-contamination routes; 
  Soaking of equipment parts in an appropriate sanitizing agent for an appropriate duration; 
  Increased frequency of cleaning and sanitizing activities for all surfaces that are not 
already cleaned or sanitized daily; 
  Heat treatment of equipment or parts, as appropriate; 
  Replacement or repair of tools, equipment, or production operation surfaces, as needed. 
Additional information on corrective action measures required and/or recommended in response 
either to positive pathogen results in spent sprout irrigation water or sprouts ( 112.148), or to 
findings of Listeria spp. or Listeria monocytogenes in your sprout production environment ( 
112.146) is discussed in Section V (Sampling and Testing of Spent Sprout Irrigation Water (or 
In-Process Sprouts)) in the draft re-issued sections (Ref. 18) and Section VI (Environmental 
Monitoring) and Section V (Seeds for Sprouting) in this document.  
IV. Agricultural Water in Sprout Operations 
In the Produce Safety Rule, we define agricultural water as water used in covered activities 
on covered produce where water is intended to, or is likely to, contact covered produce or food 
contact surfaces, including water used in growing activities (including irrigation water applied 
using direct water application methods, water used for preparing crop sprays, and water used for 
growing sprouts) and in harvesting, packing, and holding activities (including water used for 
washing or cooling harvested produce and water used for preventing dehydration of covered 
produce) ( 112.3). 
Several uses of water typical to sprouting operations meet the definition of agricultural water, 
including water used to: 
  Irrigate sprouts; 
  Prepare ice that will contact sprouts; 
  Wash finished product; 
  Wash hands during and after harvest activities; and  
  Contact food contact surfaces (FCSs) (including water used to prepare seed treatments, or 
to rinse, wash, or soak seeds prior to sprouting).  
As stated above, food contact surfaces means those surfaces that contact human food and 
those surfaces from which drainage, or other transfer, onto the food or onto surfaces that contact 
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the food ordinarily occurs during the normal course of operations. Food contact surfaces 
includes food contact surfaces of equipment and tools used during harvest, packing, and holding 
( 112.3). Food contact surfaces also includes FCSs of equipment and tools used in growing 
sprouts, such as those that contact seeds for sprouting (see  112.143(b)). While seeds used to 
grow sprouts are not themselves covered produce, seeds for  sprouting are considered food 
under Section 201(f) of FD&C Act and as defined in  112.3. Requirements relating to food 
contact surfaces in part 112 apply not only to surfaces that contact (or drain onto, or otherwise 
transfer onto) sprouts themselves but also to those that contact seeds used to grow sprouts. In a 
sprouting operation, FCSs include, for example: trays or drums used for sprouting; interior 
surfaces of containers used for seed rinsing, seed treatment, and pre-germination seed soaking; 
and counters that come into contact with sprouts during packing and/or packaging.   
It is possible that not all of your water uses meet the definition of agricultural water. For 
example, water that you use to wash the  exterior of your delivery vehicle would not be 
considered agricultural water (assuming that the vehicle  exterior does not contact the sprouts, 
and water from the exterior of the vehicle does not drain or transfer to the sprouts).  
This section of the guidance is intended to help you understand and comply with some of the 
agricultural water requirements of the Produce Safety Rule, where water is intended or likely to 
contact sprouts and FCSs. Here, we provide a limited discussion of only certain provisions of 
part 112 related to agricultural water as they relate to sprout production, most notably the 
requirements in Subpart E that agricultural water must be safe and of adequate sanitary quality 
for its intended use ( 112.41); the numerical microbial quality criterion that is relevant to 
sprouting operations ( 112.44(a)); and the related requirements for agricultural water testing 
frequency ( 112.46).2 
A. Numerical Microbial Quality Criterion for Agricultural Water Used in 
a  Sprouting Operation ( 112.44(a)) 
Certain uses of agricultural water (including water used in growing sprouts) are subject to the 
microbial water quality requirement of no detectable generic E. coli per 100 mL prescribed by  
112.44(a). We are not aware of any commonly used agricultural water applications in sprouting 
operations that are not subject to this microbial water quality requirement. Agricultural water 
uses in sprouting operations that are subject to this standard include instances where water is: 
  Used as sprout irrigation water ( 112.44(a)(1)); 
  Applied in any manner that directly contacts covered produce during or after harvest 
activities (e.g., water that is applied to covered produce for washing or cooling  activities, 
such as at entry to a wash tank, and water that is applied to harvested crops to prevent 
 
2 In the Federal Register on December 6, 2021, we issued a proposed rule that, if finalized, would amend certain 
pre-harvest agricultural requirements for covered produce (other than sprouts) (86 FR 69120). To ensure that 
interested parties can readily view the proposed pre-harvest agricultural water revisions, we also proposed to 
reorganize and replace subpart E in its entirety. Of note, this proposed rule would not substantively alter the 
standards established in part 112, subpart E, for agricultural water used for sprouts, for which the compliance dates 
have passed, or for agricultural water used during harvesting, packing, and holding activities, or for treatment of 
agricultural water. 
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dehydration before cooling), including when used to make ice that directly contacts 
covered produce during or after  harvest activities ( 112.44(a)(2)); 
  Used to contact FCSs of equipment and tools used in growing, harvesting, packing and 
holding activities of sprouts (see  112.143(b)), or to make ice that will contact FCSs ( 
112.44(a)(3)); and 
  Used for washing hands (i.e., during and after harvest activities ( 112.44(a)(4)), and 
supplied to hand-washing facilities at sprouting operations, including water used for 
hand-washing during growing activities (see  112.130(a) and (b)(2); see also  
112.143(a))). 
For sprout operations,  112.143(a) requires that growing (as well as harvesting, packing, and 
holding) take place in a fully-enclosed building. Sections 112.130(a) and (b)(2) require that, for 
growing that takes place in a fully-enclosed building, adequate and readily accessible hand-
washing facilities must be provided and must be furnished with water that satisfies the  
112.44(a) microbial quality criterion. The rule requires that water used for hand washing during 
all phases of sprout operations must meet the  112.44(a) microbial quality criterion. 
Sprout operations use different types of growing units, such as rotating drums, bins, tanks, and 
racks of trays. These growing units may use different methods to irrigate the sprouts, such as 
overhead intermittent sprays. Water for all of the irrigation methods used to irrigate sprouts must 
meet the microbial quality criterion in  112.44(a), and related requirements elsewhere in 
Subpart E ( 112.44(a)(1)).  
After sprouts are removed from the growing unit, some sprout operations wash sprouts with cool 
water to remove hulls and/or to help lower the temperature of the sprouts. Some operations use a 
bubbling water bath to loosen and float off hulls. Occasionally, sprouts are washed in 
recirculating water in a flume system. The water used in all of these applications must meet the 
microbial quality criterion in  112.44(a), and related requirements elsewhere in Subpart E. 
There are also additional requirements at  112.48 for maintaining and monitoring the quality of 
water used during harvest, packing, and holding activities. 
Ice may also be used during packing and shipping. Water used to make such ice must meet the 
standard prescribed by  112.44(a)(3). If using ice, you should take steps, such as physical 
separation, to prevent melting ice from spreading from one production batch of sprouts to other 
production batches of sprouts, or from other products to sprouts, as it may serve as a source or 
route of contamination. Additionally, water used for cleaning FCSs used for packing and holding 
sprouts must meet the standard in  112.44(a)(3).  
B. Agricultural Water Source Type and Treatment Status  
You must identify the type(s) of agricultural water source(s) used in your operation (e.g., ground 
water, water from a public water supply), and must inspect your agricultural water sources and 
distribution systems, to the extent that they are under your control, to identify conditions that are 
reasonably likely to introduce known or reasonably foreseeable hazards into or onto covered 
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produce or FCSs in light of your covered produce, practices, and conditions ( 112.42(a)). You 
must adequately maintain these systems ( 112.42(b)). 
Understanding the types of agricultural water sources that you use will help you to determine 
which requirements of Subpart E are applicable. As explained above, we are not aware of any 
uses of agricultural water (as defined in  112.3) common in sprouting operations that are not 
subject to the microbial quality requirement in  112.44(a). Therefore, our discussion below 
focuses on  112.44(a) and related provisions.  
You may not use untreated surface water for purposes covered by  112.44(a), such as for sprout 
irrigation water (see  112.44(a)). The definition of surface water appears in  112.3 and 
includes, for example, water from rivers. 
You may use untreated ground water for purposes covered by  112.44(a). The definition of 
ground water appears in  112.3 and includes, for example, water from wells that does not 
meet the definition of surface water (e.g., wells that are not influenced by surface water). In 
addition to all other applicable requirements of Subpart E that apply to agricultural water 
generally, if you use untreated ground water for purposes covered by  112.44(a), you must: 
  Sample and test water from each untreated ground water source used for such purposes, 
at the frequency established in  112.46(c), and in compliance with the methodology 
requirements established for both sampling and testing in  112.47(b) and 112.151 (see 
also Section IV. E (Agricultural Water Testing Frequency, Sampling, and Test Methods) 
below); 
  If the microbial quality criterion (no detectable generic E. coli in 100 mL) is not met, 
immediately discontinue using water from the affected water source and/or distribution 
system for any purpose covered by  112.44(a), and take appropriate corrective measures 
before using the affected water source and/or distribution system again for any such 
purpose ( 112.45(a)); and 
  Maintain records related to agricultural water testing, corrective actions, and test 
methodology, as required by  112.50(b)(2), (6), and (9). For example, if you determine 
that water you use for a purpose listed in  112.44(a) does not meet the microbial quality 
criterion established in that section,  112.45(a) requires that you take certain  steps as a 
result, and  112.50(b)(6) requires you to keep records documenting the  steps that you 
took. 
You may use treated water (i.e., water that is treated in accordance with  112.43) for purposes 
covered by 112.44(a). This is true regardless of the source from which the water was taken.  For 
example, you may use water that originated from a surface water source and has been 
subsequently treated in accordance with the requirements of  112.43. Water that has been 
treated in accordance with the requirements of  112.43 is not required to be tested to ensure 
compliance with the microbial quality criterion ( 112.46(a)(3)). In addition to all other 
applicable requirements of Subpart E that apply to use of agricultural water generally by sprout 
operations, if you use treated water for purposes covered by  112.44(a), you must: 
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  Comply with all applicable requirements for treating water (e.g., the treatment must be 
effective to achieve the no detectable generic E. coli per 100 mL microbial quality 
criterion ( 112.43(a)(1)), delivered in a manner that ensures it consistently meets that 
criterion ( 112.43(a)(2)), and monitored at a frequency adequate to ensure it 
consistently meets that criterion ( 112.43(b)). For example, an effective monitoring 
program when using a chemical treatment method would measure the level of active 
compound as well as those factors that may affect its activity, such as pH, temperature, 
and contact time;  
  Establish and keep documentation of scientific data or information you relied on to 
support the adequacy of the method you used to satisfy the requirements of  
112.43(a)(1) and (a)(2) for treating agricultural water ( 112.50(b)(3)); and 
  Maintain records related to such treatment ( 112.50(b)(4)). 
You also may use water from a public water system or supply as described in  112.46(a)(1) and 
 112.46(a)(2) for purposes covered by 112.44(a). Such water is not required to be tested to 
ensure compliance with the microbial quality criterion ( 112.46(a)(1) and (2)). In addition to 
all other applicable requirements of Subpart E that apply to agricultural water generally, if you 
use water from a  public water supply for purposes covered by  112.44(a), you must: 
  Maintain annual documentation of the results or certificates of compliance from the 
public water system or supply that demonstrate that the water meets the microbial quality 
criterion of  112.44(a) (i.e., no detectable generic E. coli per 100 mL) ( 112.50(b)(7)). 
If you hold water received from a public water system or supply in a covered tank indoors, or 
otherwise protected from potential contamination from the environment, it can continue to be 
considered public water and the applicable requirements from Subpart E described above apply.   
If you hold water received from a public water system or supply in a pond, an uncovered tank 
outdoors where it is open to the atmosphere and therefore exposed to potential contamination in 
a manner similar to the exposure of other surface water sources, the water becomes subject to the 
requirements and restrictions related to a surface  water source. For example, the prohibition 
from using untreated surface water for uses covered by  112.44(a) would apply to such water. 
If you use water received from a public water system or supply to recharge a well or otherwise 
expose the water to potential contamination in a manner similar to that of ground water sources, 
it becomes subject to the requirements related to a ground water source. For example, the 
testing requirements in  112.46(c) applicable to untreated ground water would apply to such 
water. 
C. Safe and of Adequate Sanitary Quality for Its Intended Use ( 112.41) 
In addition to the specific numerical microbial water quality criterion in  112.44(a) that applies 
to sprout operations as discussed above, a general agricultural water quality requirement applies. 
Section 112.41 requires that all agricultural water must be safe and of adequate sanitary quality 
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for its intended use. The principle of safe and of adequate sanitary quality for its intended use 
contains elements related both to the attributes of the source water and how it will be used.  
A test result indicating the agricultural water does not meet the applicable microbial water 
quality requirement in  112.44(a) demonstrates that the water is not safe or of adequate sanitary 
quality for use as agricultural water in a sprout operation. However, the converse is not 
necessarily true. That is, agricultural water that was tested and found to meet the  112.44(a) 
microbial quality criterion may not be safe or of adequate sanitary quality for such use if, for 
example, pathogens are present in the water.  For example, when conducting a routine 
inspection, under  112.42, of a well that you use as a source of untreated ground water for 
sprout irrigation, you might find a dead animal in the well. You would now have reason to 
believe that the agricultural water from that well is not safe or of adequate sanitary quality for its 
intended use. If you have determined, or have reason to believe, that your agricultural water 
source is not safe or of  adequate sanitary quality for its intended use as required by  112.41, 
you must immediately discontinue that use(s), and before you may use the agricultural water 
source and/or distribution system again for that use(s), you must take appropriate corrective 
actions, as set forth in  112.45(a). 
D. Reuse of Sprout Irrigation Water 
We recommend against re-using the spent sprout irrigation water collected from one production 
batch to irrigate subsequent batches of sprouts (e.g., applying spent water from batch 1 to irrigate 
batch 2) without any form of water treatment in between batches. The conditions that are used to 
produce sprouts allow microorganisms, including those of public health significance, to grow. 
Using spent sprout irrigation water from one production batch of sprouts to irrigate another 
production batch of sprouts can lead to cross contamination between batches, increasing the 
amount of product exposed to any contamination that may be present. Moreover, depending on 
the circumstances, such water may not be safe and of adequate sanitary quality for its intended 
use, in which case the use would be prohibited by  112.41 (Ref. 30 and Ref. 41). 
Separate from the requirements for agricultural water under Subpart E, we note that the 
requirements in Subpart M for testing spent sprout irrigation water for Salmonella spp., E. coli 
O157:H7 (and any other pathogen meeting the requirements of  112.144(c)) apply to each 
individual production batch of sprouts ( 112.147). Re-using spent sprout irrigation water from 
one production batch of sprouts to irrigate another production batch of sprouts does not relieve 
you of the requirement to conduct testing in accordance with  112.147 for spent sprout 
irrigation water from each individual production batch of sprouts (see Section V (Sampling and 
Testing of Spent Sprout Irrigation Water (or In-Process Sprouts)) in the draft re-issued sections 
(Ref. 18)). 
Similarly, we also recommend that you not reuse spent sprout irrigation water to irrigate the 
same production batch of sprouts over time without any form of water treatment in between uses. 
Reusing spent sprout irrigation water for subsequent irrigation of the same production batch of 
sprouts over time could reintroduce any pathogens present in the water into the growing sprouts, 
which could contaminate a greater proportion of a given batch of sprouts. Any pathogens that 
may be present in the reused spent sprout irrigation water could then be added to the growing 
sprouts and could multiply to higher numbers during sprouting (Ref. 42).   
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E. Agricultural Water Testing Frequency, Sampling, and Test Methods 
You are required to test your agricultural water source(s) to ensure compliance with the 
microbial quality criterion in  112.44(a) if you use untreated ground water as agricultural water 
(see Section IV. B (Agricultural Water Source Type and Treatment Status) above). In this 
circumstance, you are initially required to test  each such water source a minimum of four times 
throughout the growing season or over a period of  one year (112.46(c)). If all four of the initial 
samples tested meet the microbial quality criterion in  112.44(a), you may test once annually 
thereafter. You must resume testing at least four times per  growing season or year if any annual 
test fails to meet the microbial quality criterion in  112.44(a). 
Section 112.46(c) requires all such samples to be representative of the intended use(s). By 
representative of the intended use(s) we mean collected from a location, and at a time of year, 
such that the sample can reasonably be expected to represent the quality of the agricultural water 
as it is normally used. For example, if you normally use untreated ground water for agricultural 
water uses only from April through August of each year, you should collect samples during that 
part of the year so that the samples are representative of the intended use(s). You should assess 
your production schedule to determine an appropriate sampling scheme for collecting the four 
initial samples. 
With respect to sampling location, sampling at the point of your actual use is ideal (e.g., 
sampling where water comes out of a faucet or hose in your building and enters your sprout 
growing unit to be used for irrigation). You may also sample at other points along the water 
distribution system, from the water source itself to the point of use, as long as there is no 
reasonably likely point of contamination in the water distribution system between your chosen 
sampling point and the point of use, such that the sample can reasonably be expected to represent 
the quality of the water when it is used. For example, if untreated ground water is drawn from a 
well that is not under your control, you might  choose to collect samples from a point that is 
under your control, such as a valve on a pipe at the edge of your property line. As another 
example, if the well is under your control and the piping from the well head to the points of use 
is in good condition, you may choose to collect samples from the well head. In either case, such 
samples can be considered representative of the intended use(s) (with respect to the location of 
the samples), provided that there is no reasonably likely point of contamination in the water 
distribution system between your chosen sampling point and the point of use. Further,  
112.42(a) requires you to conduct an inspection of your agricultural water system, to the extent 
under your control, to identify conditions that are reasonably likely to introduce known or 
reasonably foreseeable hazards. This inspection is required at the beginning of a growing season, 
as appropriate, but at least annually. This inspection should assist you in selecting your sampling 
points by determining what sampling locations are, or are not, representative of your intended 
use(s). Section 112.50(b)(1) requires you to establish and keep records of the agricultural water 
system inspection findings required by  112.42(a). 
Agricultural water samples to be tested for compliance with the  112.44(a) microbial quality 
criterion must be collected aseptically, in accordance with  112.47(b). You must use sterile 
equipment and tools and aseptic technique when collecting samples. Note that cleaning and 
sanitizing of sampling equipment and tools may not result in their being sterile (See Appendix 1 
(Aseptic Sampling)). 
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Agricultural water samples must be analyzed for compliance with the  112.44(a) microbial 
quality criterion using a method set forth in  112.151 ( 112.47(b)). We reviewed publicly 
available test methods for water and determined that EPA Method 1603 is appropriate for testing 
water quality for this purpose ( 112.151(a)). 
However, we recognize that other scientifically valid analytical methods may be available or 
may become available in the future. Therefore, we provide flexibility for you to use any other 
scientifically valid analytical method that is at least equivalent to the prescribed analytical 
method (i.e., EPA Method 1603) in accuracy, precision in detecting the relevant organism or 
indicator (i.e., generic E. coli) and precision in the relevant sample matrix (i.e., ground water), 
and sensitivity ( 112.151(b)(1)). We have identified a number of testing methodologies that 
meet the requirements of  112.151(b)(1), and have posted a list of these methods on our 
website. See Equivalent Testing Methodologies for Agricultural Water at 
https://www.fda.gov/food/laboratory-methods-food/equivalent-testing-methodology-agricultural-
water.   
Section 112.50(b)(9) requires you to establish and keep documentation of any analytical methods 
that you choose to use for agricultural water testing in lieu of the method that is incorporated by 
reference in  112.151(a). We recommend that records established and kept in compliance with  
112.50(b)(9) include 1) name and identification number of the method validated by third-party 
methods validation organization, or 2) detailed analytical procedures, an explanation of why the 
alternate method is at least equivalent to the reference method (such as a scientific study, or data 
or information regarding the sensitivity, specificity, and precision of the alternate method 
compared to the reference method), and any other relevant information supporting the use of the 
alternate method. We note that you are not required to notify us or submit information about such 
methods of analysis for our review or approval prior to use.  
You should choose a laboratory that is qualified to test agricultural water for generic E. coli. 
Testing is typically contracted to a third-party testing laboratory; however, testing may also be 
performed by a sprout operations own laboratory (e.g., an operations own in-house 
laboratory). Using an accredited laboratory (e.g., a laboratory accredited to International 
Organization for Standardization (ISO) Standard 17025) is one way to have confidence that the 
laboratory will provide reliable, accurate test results.   
F. Post-Harvest Water Management 
Section 112.48(a) requires you to manage the water used during harvest, packing, and holding 
activities for sprouts, as necessary, including by establishing and following water-change 
schedules for re-circulated water to maintain its safety and adequate sanitary quality and 
minimize the potential for contamination of covered produce and FCSs with known or 
reasonably foreseeable hazards. Section 112.48(b) requires you to visually monitor the quality of 
water that you use during harvest, packing, and holding activities. For example, as a sprout 
operation:  
  You must visually monitor water used for washing and cooling sprouts for buildup of 
organic material (such as hulls and plant debris). We recommend against the reuse of 
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untreated post-harvest water in multiple production batches of sprouts because of the 
potential risk associated with its use.  
  You should consider using a water treatment system as part of managing your post-
harvest water. If you choose to treat your water, those treatments should be applied in a 
manner that is consistent with  112.43.  
  The specific level and frequency of post-harvest water treatment, the point at which 
treatment should be applied, and the time intervals for monitoring treatments necessary to 
align with  112.43 may vary. An effective monitoring program for use of a chemical 
treatment method should measure the level of active compound as well as those factors 
that may affect its activity, such as pH, temperature, contact time, and organic load. For 
example, adequate monitoring of water treated with hypochlorite in a postharvest wash 
for sprouts should include monitoring the level of active antimicrobial (free available 
chlorine) and pH, since hypochlorite activity is reduced both by organic material (e.g., 
soil, plant debris) and pH values outside its effective range (pH 6.07.5) (Ref. 43, Ref. 
44, Ref. 45, and Ref. 46).  
V. Seeds for Sprouting 
The requirements in  112.142 are designed to reduce the likelihood that seeds for sprouting 
serve as a vehicle for introducing contamination in sprouts. This section of the guidance provides 
recommendations to help you comply with these requirements for receiving, storing, and treating 
seeds for sprouting. As previously mentioned, throughout this guidance, for the ease of the 
reader, we often refer collectively to everything sprouted to produce sprouts for human 
consumption, including beans, simply as seeds. In the Rule, we used the phrase seeds or 
beans to remove any potential confusion as to whether beans for sprouting were included. 
References to seeds in this guidance should not be read to exclude other things that are 
sprouted to produce sprouts for human consumption, such as beans or nuts. 
While poor sanitation and unhygienic practices at the sprout operation can contribute to 
contamination of sprouts, studies indicate that contaminated seed is the likely source of most 
sprout-related outbreaks (Ref. 2, Ref. 11, Ref. 47, and Ref. 48). Seed contamination can occur at 
various points in the supply chain, including the seed farm, seed conditioner, seed supplier, 
transportation, or the sprout operation. The sprout growing process provides opportunities for 
contamination that may be present in or on a few seeds to spread through the entire sprout 
production batch. 
Only a small portion of seed produced in the United States is destined for sprouting, while seed 
is more frequently used as planting stock for forages for livestock or for field cultivation of 
human food (Ref. 2). Although we encourage you to purchase seeds that were produced 
specifically for sprouting and grown under Good Agricultural Practices (GAPs) (i.e., FDAs 
guidance titled Reducing Microbial Food Safety Hazards in the Production of Seed for 
Sprouting) (Ref. 17), we are aware that this does not always occur. For this reason, seeds that 
you use for sprouting may be grown, milled, and/or stored under conditions where contamination 
is likely to occur. Seed may become contaminated by potential sources of fecal contamination, 
such as contaminated water, inadequately treated or raw manure used as fertilizer, wild or 
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domesticated animals, unclean equipment, or inadequate worker hygiene. These conditions may 
contribute to the presence of human pathogens on seed for sprouting. 
The steps of sorting, cleaning, storing, packaging, and shipping of seeds by seed mills may 
spread contamination between seed lots or within a seed lot, especially if GAPs are not followed. 
During these steps, seeds from multiple lots of different origins may be mixed together, 
providing opportunity for cross contamination between lots to occur, and complicating any 
corrective actions and/or traceback of the origin of the lots should they be involved in a 
contamination event or illness (Ref. 2). Use of unclean equipment may introduce contamination 
to a seed lot or spread contamination to other seed lots if the same equipment is used with 
multiple lots and not cleaned in between uses with different seed lots. Damage to the seeds 
during these steps may also affect the overall safety of the seeds; such damage may occur 
inadvertently or deliberately (i.e., scarification). Cracks, crevices, and other types of damage to 
seeds may facilitate the internalization of pathogens into seeds and make it difficult to remove 
pathogens during subsequent treatment and handling (Ref. 2). 
A. Seed Sourcing, Receiving, Handling and Storage 
1.  Seed Sourcing 
You must take measures reasonably necessary to prevent the introduction of known or 
reasonably foreseeable hazards into or onto seeds that you will use for sprouting ( 112.142(a)). 
We are aware that some sprout seed suppliers seek written assurances from their seed growers 
and handlers that their seeds were produced under GAPs and handled according to food safety 
best practices throughout harvesting, conditioning, storage, and transportation. Sprout operations 
can request this information from the entity which sells their seeds. While not requirements of 
the Produce Safety Rule, we recommend these practices as prudent when sourcing the seeds used 
for sprouting. In addition, we are aware that production of seed for sprouting under GAPs is 
required by some foreign governments (Ref. 21) and is accepted as an international food safety 
best practice (Ref. 19). We have published guidance titled, Reducing Microbial Food Safety 
Hazards in the Production of Seed for Sprouting (Ref. 17) which recommends GAPs for seed 
growers and handlers to produce seed for sprouting in accordance with food safety practices. We 
recommend that you source seeds for sprouting that have been grown, packed, and held 
according to this guidance. 
Knowing your seed supplier(s) and establishing specifications for the seeds you receive (such as 
being grown under GAPs and handled under sanitary conditions during storage and distribution 
or transport) can help improve the safety of the seeds you receive. Specifications also provide 
standards against which you can assess the acceptability of the seeds you receive for the 
production of finished sprouts. 
We further recommend having a proactive discussion with your seed supplier to learn what its 
practices are in the event that a portion of a master lot of seed tests positive for a pathogen prior 
to distribution (e.g., does the supplier divert the entire master lot to a use other than seeds for 
sprouting?). You should also ask whether seed from different sources is generally commingled 
before a master lot is broken down into sub-lots, to determine the relevance of any future 
positive test results on multiple sub-lots. We also recommend having a discussion with your seed 
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supplier about how you should notify them of any positive test results you receive (e.g., of spent 
sprout irrigation water as required under  112.142(b)(2)), including what information they 
would need and under what circumstances they would accept a return of a seed lot that had tested 
positive for a pathogen (e.g., results of confirmatory testing). It is prudent to obtain this 
information up front, rather than waiting to obtain it after receiving a positive test result. We also 
recommend you ask the seed supplier what actions they take when notified of contamination 
(e.g., by regulators or other sprout operations) associated with a seed lot that has already entered 
commerce.  
2.  Seed receiving 
We recommend you develop a seed receiving program setting out the standard operating 
procedures (SOPs) you follow to receive and inspect seed upon receipt and the criteria you use in 
determining whether to accept a shipment. The seed receiving SOP should describe the 
objectives, description of procedures for receiving seeds, including parameters that should be 
monitored or verified (e.g., in the form of a checklist), documentation that should be received, 
who is responsible for each task, materials or supplies needed (e.g., black light), and actions to 
be taken in the event that non-conformities are identified. An example of a seed receiving and 
inspection checklist can be found in the next section (Section V. A. 3 (Visual Inspection of Seeds 
and Their Packaging)). Your seed receiving program should include specific procedures for how 
you will handle any issues identified while inspecting a shipment of seed.   
Once seeds arrive at your operation, you should verify that the transport vehicle is clean and 
sanitary. You should also check the seed tag, package labeling and other documentation to 
ensure that the seeds are what you ordered (e.g., they match the information on the purchase 
order), that they are from a known supplier, that they meet any specifications you may have 
established with your seed supplier, and that any information or documentation that you plan to 
keep for your own records (such as documentation of the seed lot number, or of prior treatment 
of seed) has been provided. If requested information or documentation pertinent to the seed lot is 
missing, you may consider following up with your seed supplier to obtain that information. If a 
shipment does not meet your requirements, you should evaluate the deviation(s) and, based on 
your criteria for accepting seeds, determine whether the safety of the seed may be compromised 
and whether the lot should be rejected. You should be able to account for lots rejected due to 
evidence of contamination or for other reasons (e.g., in the event of a future recall). 
You should not use seed that is labeled Not for Human Consumption for growing sprouts for 
human consumption, as these seeds may have been grown, handled, or treated in such a way that 
makes them inappropriate for human food. As you are sourcing seed, we recommend asking your 
seed supplier if the lot of seed you are purchasing has tested positive for a pathogen or if the seed 
lot came from a larger master lot, and if so, whether any test done on any other part of that 
master lot was positive for a pathogen. We recommend against using seed from sub-lots of a 
master lot that has tested positive for a pathogen unless you have information that the source of 
the contamination to the part of the lot that tested positive is not applicable to your sub-lot (see 
also Section V. A. 4 (Seed Testing), below, for a discussion of  112.142(c)). This is because of 
the likelihood that other portions of the master lot could also be contaminated, and the limitations 
of seed testing methods to detect sporadic contamination.  
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3.  Visual inspection of seeds and their packaging 
You must visually examine seeds, and packaging used to ship seeds, for signs of potential 
contamination with known or reasonably foreseeable hazards ( 112.142(d)). Each bag should be 
examined for physical damage that could result in contamination with such hazards (e.g., holes 
from rodents) and other signs of contamination (e.g., stains, insects, presence of insect, bird, or 
rodent feces, presence of rodent urine, or foreign material) upon arrival. This can be as simple as 
using a black light in a dark room to examine seed packaging for signs of rodent urine, and 
visually examining the seeds and packaging for rodent feces, rodent gnawed holes, or excessive 
dirt, debris, and damage. In addition to looking for signs of rodent damage, you should also look 
for damaged seeds. Damage to seeds can provide small, hard to reach places for pathogens to 
lodge in and make any treatment to reduce pathogens less effective. We also recommend that if 
you store the seeds for any length of time after they arrive at your operation, you should re-
inspect the bags of seeds for signs of contamination before using them for sprout production. 
Procedures for receiving and inspecting seed (e.g., that could be included in a seed receiving and 
inspection checklist) should include the following information included in Table 2.  
Table 2. Suggested Procedures for Receiving and Inspecting Seed 
Check the transportation vehicle for conditions that could lead to contamination; 
Verify supplier name and address on packages to see if they match those on the purchase 
order; 
Check date of shipping and volume of products shipped; 
Verify conformance to your seed specifications (e.g., seed type, origin, grown in accordance 
with GAPs for seeds (i.e., FDAs guidance titled Reducing Microbial Food Safety Hazards 
in the Production of Seed for Sprouting) (Ref. 17), prior seed treatment); 
Check lot information was provided (e.g., lot size, lot number, master lot number (if 
applicable), name of establishments from which seed originated as well as establishments that 
handled seed, harvest date); 
Check condition of bags of seed (e.g., water damage, off odor, presence of insect, bird, or 
rodent feces, presence of rodent urine using a blacklight, mechanical damage, mold), 
ensuring that packaging does not allow for contamination and is intact; 
Check seed testing record, or letter of guarantee or certificate of conformance for such 
testing, if available, including lot number tested, name of laboratory, test results, and details 
of tests performed (e.g., test method used, information to support scientific validity of test 
method, date tested);  
Check seed treatment record, or letter of guarantee or certificate of conformance for such 
treatment, if available; and   
Account for lots rejected due to evidence of contamination or for other reasons. 
 
If you observe obvious signs of seed contamination upon receipt and you conclude the 
contamination occurred prior to the seed arriving at your sprout operation, you should report the 
information to the seed grower, distributor, supplier, or other entity from whom you received the 
seed, and return the shipment to the supplier. You should also discontinue use of the affected part 
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of the seed lot, and we recommend that you conduct intensified cleaning and sanitizing of any 
surfaces that may have become contaminated by the seed lot (see Section III (Cleaning and 
Sanitizing)) and take any other actions necessary to prevent reoccurrence of contamination. If 
you know or have reason to believe that a lot of incoming seeds may be contaminated with a 
pathogen, either because of a microbial test result or because it has been associated with 
foodborne illness, there are steps that you must take ( 112.142(b) and (c)), and you must 
document those actions as required under 112.150(b)(6). These steps are discussed in the 
section of this guidance document, Section V. C (Corrective Actions for Seeds That May Be 
Contaminated with a Pathogen). 
4.  Seed testing 
The Produce Safety Rule does not require microbial testing of seeds for sprouting by you or your 
supplier. However, if you or your supplier choose to conduct seed testing, we recommend that 
you use a sampling plan that results in a sample that is representative of the entire lot of seed, 
collect the sample aseptically, and test the sample using a scientifically valid method. Because 
microbial contamination in seeds, when present, is often at low levels and not uniformly 
distributed throughout a seed lot, such contamination is often difficult to detect (Ref. 2). If you or 
your seed supplier test a lot of seeds for pathogens, you should be aware that the absence of a 
positive test result for pathogens does not mean that the lot of seeds is necessarily pathogen-free.  
A positive test result for a pathogen indicates that the lot of seeds is contaminated. In such a case, 
you must take certain actions under  112.142(b), including discontinuing use of that seed lot for 
sprout production, except as provided for in certain limited circumstances in  112.142(c), and 
reporting the information to the seed grower, distributor, supplier, or other entity from whom you 
received the seeds. One such circumstance is if you treat the lot of seeds with a process that is 
reasonably certain to achieve destruction or elimination of the most resistant microorganisms of 
public health significance that are likely to occur in the seeds ( 112.142(c)(1)). We understand 
that some in the food industry may refer to such a treatment as a pasteurization step. However, 
we note that processes that meet the description in  112.142(c)(1) are not currently commonly 
used in the sprouting industry, in part, because such processes would be likely to impact the 
germination ability of the seed. Such processes are far more robust than seed treatments 
described in  112.142(e), which typically only reduce the numbers of microorganisms of public 
health significance, rather than eliminate them. For more information, see the seed treatment 
section (Section V.B (Seed Treatment)) of this document below. If you choose to conduct such a 
treatment (i.e., pasteurization step) on your seed lot so that you may use those seeds for 
sprouting, we note that you must still comply with the requirement in  112.142(b)(2) to report 
the positive test result to your seed grower, distributor, or supplier. 
You are not required to take the steps set forth in  112.142(b)(1) and (2) if you later reasonably 
determine, through appropriate follow-up actions, that the lot of seeds is not the source of 
contamination ( 112.142(c)(2)). However, we consider that there are very few circumstances in 
which it would be appropriate to apply the exception at  112.142(c)(2) in the event there has 
been a positive pathogen test result in an incoming lot of seed. 
It is also important to note that if a positive pathogen test result is obtained from a seed lot, re-
testing a new sample from the same seed lot and obtaining a negative pathogen test result does 
31 
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not negate the previous positive test result. In such a case, the required follow-up actions must 
still be taken.  
5.  Seed storage 
Seeds can become contaminated during storage. Storing seed for sprouting is considered part of 
the process of growing sprouts, and therefore the building in which your seed is stored is subject 
to the applicable requirements for buildings in Subpart L ( 112.122(a)). Further, seed for 
sprouting must be stored in a fully-enclosed building (112.143(a)), which should be of sound 
construction and in good repair. You must take measures reasonably necessary to prevent the 
introduction of known or reasonably foreseeable hazards into or onto seeds that you will use for 
sprouting ( 112.142(a)). Many pests are attracted to seeds, and these pests can serve as a source 
of, or vector for, spreading contamination, especially when seeds are in storage. In order to 
prevent the introduction of known or reasonably foreseeable hazards into or onto seeds that will 
be used for sprouting, you should store seeds in a protected manner. Your seed storage area 
should be clean, dry, and dedicated to seed storage. You must take measures to exclude pests 
from the building ( 112.143(a), 112.126, and 112.128(b), and see Section IV (Equipment, 
Tools, and Buildings) in the draft re-issued sections (Ref. 18)) and you must routinely monitor 
for pests, as necessary and appropriate ( 112.128, and see Section IV (Equipment, Tools, and 
Buildings) in the draft re-issued sections (Ref. 18)). You should store seeds off the floor and 
away from walls to facilitate pest control inspection. You should regularly inspect seed 
packaging and containers (see Section IV (Equipment, Tools, and Buildings) in the draft re-
issued sections (Ref. 18)). 
Once the original seed packaging is opened, the remaining seed must be stored in closed 
containers with tight-fitting lids or otherwise protected from contamination ( 112.142(a)). If you 
use containers other than the original packaging to hold seeds, their food contact surfaces (FCSs) 
must be cleaned and sanitized prior to use (see  112.143(b)). Containers should also be labeled 
to maintain the identity of the seed lot they contain. If the seeds have received prior antimicrobial 
treatment by a grower, supplier or distributor, this should be indicated on the label. 
If you intend to rely solely on a seed treatment conducted by an entity earlier in the supply chain 
(e.g., you will not re-treat the seeds yourself), you must store seeds that have received that prior 
treatment by the seed grower, distributor, or supplier under conditions that will prevent them 
from becoming contaminated (see  112.142(e)(2)(ii)). We recommend that seeds that received 
prior treatment be stored separately from untreated seeds, in airtight containers/packages that are 
as small as practicable to minimize the number of times any particular container/package is 
opened and closed to remove seed from that container (because each such event presents the 
possibility of contamination). If you reuse containers/packaging, you must clean and sanitize the 
FCSs on these containers before contact with seeds used to grow sprouts ( 112.143(b)).  
B. Seed Treatment 
Research indicates that seed contamination, when it occurs, may be at low levels, intermittent, 
and unequally distributed within seed lots (Ref. 2). However, even low levels of pathogens on 
seed for sprouting are a concern, because the conditions present during sprouting are ideal to 
support the growth of pathogens. There is some evidence that sprout operations associated with 
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outbreaks often did not apply seed treatments correctly, consistently, or at all in the production of 
sprouts associated with the outbreak (Ref. 2). While treating seeds used for sprouting does not 
guarantee pathogen-free sprouts, seed treatment has been shown to reduce the percentage of 
contaminated batches of sprouts (Ref. 49, Ref. 50, Ref. 51, Ref. 52, Ref. 53, and Ref. 54). For 
example, a quantitative risk assessment model predicts that, without any seed treatment or spent 
sprout irrigation water testing, 5.22% of sprout batches will be contaminated with Salmonella 
(Ref. 54). Following seed treatment, the predicted fraction of contaminated sprout batches was 
reduced to 2.32%, 0.0320%, and 0.000320% when 1-log, 3-log, or 5-log seed treatment was 
applied, respectively. When seed treatment was combined with spent sprout irrigation water 
testing, the fraction of positive sprout batches predicted by this model was reduced further. (This 
risk assessment is discussed in more detail below.) Therefore, seed treatment is a critical part of a 
multi-hurdle approach to reduce the public health risks associated with the consumption of 
sprouts. 
Section 112.142(e) requires that the seeds you use to grow sprouts be treated using a 
scientifically valid method to reduce microorganisms of public health significance. We use the 
term scientifically valid to mean an approach that is based on scientific information, data, or 
results published in, for example, scientific journals, references, textbooks, or proprietary 
research. We note that the term scientifically valid has a different meaning than validated 
with regard to seed treatment, for the purposes of the Produce Safety Rule. The term validation 
study is often used to refer to a systematic process of obtaining and evaluating scientific and 
technical evidence that a control measure or a combination of control measures is capable of 
effectively controlling the identified hazard(s). The Produce Safety Rule does not require you to 
perform a validation study to evaluate a seed treatment; however, we note that a validation study 
can help to evaluate whether a particular treatment is scientifically valid. 
To meet the seed treatment requirement, you must adopt one of the following two approaches: 
  Treat seeds that you use to grow sprouts using a scientifically valid method to reduce 
microorganisms of public health significance ( 112.142(e)(1)). If you perform treatment 
of seeds used for sprouting in accordance with  112.142(e)(1), you must establish and 
keep documentation of this treatment (112.150(b)(1)); or 
  Alternatively, you may rely on prior treatment of seeds conducted by a grower, 
distributor, or supplier of the seeds (whether to fulfill this requirement completely or for 
the purpose of you considering such prior treatment when applying appropriate additional 
treatment of the seeds at your operation immediately before sprouting), provided that you 
obtain documentation (such as a Certificate of Conformance) from the grower, 
distributor, or supplier that: (i) the prior treatment was conducted using a scientifically 
valid method to reduce microorganisms of public health significance; and (ii) the treated 
seeds were handled and packaged following the treatment in a manner that minimizes the 
potential for contamination ( 112.142(e)(2)). Such records should include 
documentation of the scientifically valid method used to treat the seeds, references to 
scientific data supporting the method, the level of log reduction achieved, the type of 
seeds used for any validation study that may have been done, the pathogens targeted, and 
information on how the seeds were handled and packaged following the treatment to 
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minimize the potential for contamination (e.g., seeds were packaged immediately after 
treatment, type of packaging). 
1. Choosing a seed treatment 
A successful seed treatment reduces microbial pathogens while preserving seed viability, 
germination, and vigor. Seed types can vary in sensitivity to antimicrobial agents and other types 
of treatments, which can affect how well the seeds germinate and grow after treatment. 
Additionally, the varying surface features of different types of seeds can influence how well a 
treatment can access and inactivate pathogens on or in the seed. Therefore, an antimicrobial 
treatment that is effective for one type of seed may not be as appropriate for other types. 
When reviewing the options available for seed treatment, especially if you plan to treat seeds at 
your operation (as opposed to, or in addition to, purchasing pre-treated seeds), you should 
 
consider the feasibility of applying the treatment at your operation. For example, irradiation3 is 
an option for seed treatment to reduce microorganisms of public health significance that may not 
be feasible for you to apply on-site. In addition, hot water treatments have been demonstrated to 
reduce pathogens on seeds by more than 5 log CFU/g in one study (Ref. 55) and to undetectable 
levels in another study (Ref. 56). However, these treatments can require use of equipment, such 
as industrial-sized hot water pasteurization machines (Ref. 57), that might be cost prohibitive for 
a small sprout operation. 
Nothing in the Produce Safety Rule requires or authorizes you to take measures in conflict with 
existing federal, State, or local regulations. We expect any seed treatment that is used to be 
applied in accordance with all applicable federal, state, tribal, or local requirements. For 
example, sources of radiation used to treat food are considered food additives, under the law, and 
require an authorizing regulation by FDA. Also, in compliance with the Federal Insecticide, 
Fungicide and Rodenticide Act (FIFRA), pesticide chemicals used as seed treatments must be 
registered with the Environmental Protection Agency (EPA) and labeled for use to reduce 
microorganisms of public health significance on seeds for sprouting. Unlike pesticide chemicals, 
equipment that works to control microorganisms by physical means (e.g., heat) are classified by 
EPA as pesticide devices and do not require registration with EPA under FIFRA (Ref. 58). 
Note also that some States require registration of pesticide devices, and we refer you to the 
appropriate State pesticide regulatory agency for more information on a particular States 
requirements related to such devices (Ref. 59).  
Rather than establishing a specific type or method of seed treatment that you must use to treat 
seeds, the Produce Safety Rule allows you to use any scientifically valid method to reduce 
microorganisms of public health significance on seeds that you use to grow sprouts ( 
 
3 21 CFR  179.26(b)(10) allows seed for sprouting to be treated with ionizing radiation up to a maximum dose of 8 
kGy. We note that the codified language in the regulation states seeds, but for the purposes of the regulation, the 
FDA recognizes that beans for sprouting (e.g., mung bean seeds used for sprouting) would also be included. For 
information about the use of non-ionizing sources of radiation (such as UV light, radio frequency, microwaves, and 
pulsed light) we refer the reader to 21 CFR Part  179 more generally. We note that while seeds that have been 
treated with ionizing radiation must be labeled with a radura symbol along with either the statement Treated with 
radiation or the statement Treated by irradiation, sprouts that are grown using irradiated seeds need not be so 
labeled where the sprouts themselves have not been irradiated (see 21 CFR  179.26(c)(2); 65 FR 64605, at 64606-7 
(Oct. 30, 2000)). 
34 
  Contains Nonbinding Recommendations 
112.142(e)). This approach provides you with flexibility in choosing a seed treatment. In 
choosing a treatment(s) appropriate to your operation and your sprouts, and in compliance with 
the Produce Safety Rule, there are a number of things to consider. In this guidance, we highlight 
certain treatments that have been studied in the literature and discuss important considerations 
related to using those treatments. 
Known seed treatment methods include those that work by chemical means (e.g., liquid or gas), 
physical means (e.g., heat), or a combination of these. Based on a review of available literature, 
physical and combination type treatments have been reported to be the most effective for 
reducing the numbers of pathogens on seeds for sprouting. Physical treatments, such as heat (dry 
heat or hot water), high pressure, and irradiation are reported to have better penetration 
characteristics for reaching bacteria on microscopically rough surfaces, as well in as the interior 
of the seed, compared to chemical treatments (Ref. 60). In some studies, physical treatments 
have been reported to achieve a 5-log or greater reduction in pathogens on seeds (Ref. 57, Ref. 
60, Ref. 61, and Ref. 62). Combination treatments (i.e., applying two or more methods 
sequentially or simultaneously) may be more effective than using a single treatment alone. 
Examples of such combination treatments include: chemical plus heat; irradiation plus dry heat; 
and high pressure plus dry heat. Literature suggests many combination treatments may be able to 
achieve a 5-log or greater reduction in pathogens on seeds (Ref. 57, Ref. 60, Ref. 61, Ref. 62, 
Ref. 63, Ref. 64, and Ref. 65). 
Previous studies suggest that the following chemicals, when used at appropriate concentrations, 
may be able to achieve at least a 3-log reduction in pathogens on seeds: acidified sodium chlorite 
(Ref. 66); calcium hydroxide (Ref. 67), calcium hypochlorite4 (Ref. 60); caprylic acid (Ref. 68); 
gaseous acetic acid (Ref. 69 and Ref. 70); hydrogen peroxide (Ref. 67 and Ref. 71); lactic acid 
(Ref. 64); monocaprylin (Ref. 68); oxalic acid (Ref. 64); phytic acid (Ref. 64); and sodium 
hypochlorite (Ref. 72). This is not intended to be an exhaustive list. We note, however, that such 
treatments require EPA approval and pesticide registration under FIFRA before they may be 
used for such purposes. We refer readers to EPA for further information on the registration 
process. Information regarding currently registered pesticide products is available on EPAs 
website at:  https://iaspub.epa.gov/apex/pesticides/f?p=PPLS:1. 
2. Seed treatment efficacy 
Many seed treatments reduce, but do not eliminate, microorganisms of public health significance 
that may be present on the seeds. Pathogens that are not eliminated by seed treatment can be 
amplified during the sprouting process; therefore, adequate seed treatment prior to sprouting is a 
key component of the multi-hurdle risk reduction framework for sprouts established in the 
Produce Safety Rule.  
The use of seed treatment in a multi-hurdle risk reduction framework is supported by the 
findings of a 2018 quantitative risk assessment conducted by researchers at FDAs Center for 
Food Safety and Applied Nutrition (Ref. 54). They assessed the risk of salmonellosis associated 
 
4 In our prior Sprout Guidances, we cited a 20,000 ppm calcium hypochlorite treatment as an example of a seed 
treatment. The EPA registration for this treatment has since been updated and we refer readers to EPA for further 
information on that registration. 
35 
  Contains Nonbinding Recommendations 
with the consumption of alfalfa sprouts to evaluate the public health impact of treating seeds and 
testing spent sprout irrigation water for Salmonella cells washed off during production. If there 
were no interventions, the assessment estimates that there would be 76,000 (95% confidence 
interval (CI) 15,400-248,000)5 cases of salmonellosis every year tied to the consumption of 
contaminated sprouts. The model indicates that the risk reduction from using a seed treatment 
that can achieve a 1-log reduction in Salmonella alone (i.e., without spent sprout irrigation water 
testing) is comparable to spent sprout irrigation water testing alone, and each additional 1-log 
reduction in Salmonella achieved by the seed treatment was predicted to provide additional 
reduction (i.e., greater than spent sprout irrigation water testing alone). 
Among the approaches that the researchers evaluated, the one they found to have the greatest 
public health impact, is using a seed treatment that obtains a 5-log reduction of Salmonella 
combined with testing spent sprout irrigation water. Together, these two interventions would 
result in an estimated 99.9994% reduction in the number of salmonellosis cases per year  
essentially no illnesses (95% CI <1-1.5). A 3-log reduction treatment in combination with 
effective spent sprout irrigation water testing (i.e., collecting and testing a sample that is 
representative of 100% of your batch of sprouts) would reduce the number of cases per year to 
45 (95% CI 10-146). Both approaches provide substantial public health benefits. However, the 
researchers estimate that there would be hundreds, or thousands, of illnesses if spent sprout 
irrigation water is tested in combination with treatments that achieve less than a 3-log reduction. 
For example, testing spent sprout irrigation water with a treatment that can obtain a 1-log 
reduction of Salmonella is predicted to only reduce the number of cases per year to 3,560 (95% 
CI 821-11,400). Testing spent sprout irrigation water alone would only reduce the cases to an 
estimated 12,100 (95% CI 2,400-41,200) per year. 
Based on the results of this risk assessment, we recommend the use of a seed treatment that 
achieves at least a 3-log reduction in microorganisms of public health significance. As more 
efficacious seed treatments become readily available on the market and registered for this use, it 
is likely that we will update this recommendation in the future. For more information on spent 
sprout irrigation water testing, see Section V (Sampling and Testing of Spent Sprout Irrigation 
Water (or In-Process Sprouts)) in the draft re-issued sections (Ref. 18). 
We recommend that you use the most efficacious seed treatment available that is practical for 
your operation to reduce the presence of microorganisms of public health significance on seeds 
for sprouting, with primary consideration given to reduction of Salmonella spp. and E. coli 
O157:H7. You should not rely only on a treatment that achieves a low level of reduction if other, 
more effective, treatments that are practical for your operation are available. We understand that 
a 3-log reduction is the minimum level of reduction of pathogens the EPA will consider when 
registering an antimicrobial treatment for seeds that includes a public health claim. Because 
using a treatment that provides a higher level of pathogen elimination before spent sprout 
irrigation water testing is undertaken is expected to reduce the number of production batches of 
sprouts that test positive for a pathogen (Ref. 54), such an approach would minimize the 
likelihood that you will invest time and resources to produce a production batch of sprouts that is 
 
5 The confidence interval (CI) numbers are the lower and upper range of the estimate. The estimates in this risk 
assessment do not factor in interventions that industry is currently conducting; they are designed to compare the 
impact of different approaches.  
36 
  Contains Nonbinding Recommendations 
later determined to be contaminated through routine testing required under  112.147 (and which 
must be discarded as a result, as required by  112.148).   
The validity and efficacy of any treatment is dependent on the specific parameters governing 
how the treatment is applied. You should carefully monitor those variables that impact the 
overall efficacy of the treatment, including, for example, seed type, treatment concentration, 
treatment time, temperature, pH, pressure, radiation dose, seed-to-treatment solution ratio, pre- 
and post-treatment rinsing, and agitation (as appropriate to maximize contact between seed and 
the treatment solution or gas). You must document observations of treatment and key parameters 
monitored during preparation and application of treatments (e.g., chemical composition of 
treatment solution, heating temperature, equipment used, concentration of treatment solution, 
treatment time, pH of treatment solution, presence of agitation)) ( 112.161)(a)(1)(ii)). If you are 
utilizing a seed treatment described in scientific literature, you should ensure that your use of the 
treatment is within the parameters used in the study. In other words, you should ensure that the 
conditions under which the treatment was applied in the study are compatible with how you will 
be applying the treatment at your operation. For example, if the treatment you are considering 
has only been tested on alfalfa seeds, and you plan to use the treatment on mung beans, you 
should not assume that the treatment will be equally effective at reducing microorganisms of 
public health significance on mung beans. You should, instead, obtain a study that was 
conducted on the type of seeds that you intend to treat.  
The Produce Safety Rule does not prohibit the use of proprietary seed treatments (e.g., 
treatments developed based on a firms own scientific research not published in scientific 
literature). However, your use of such a treatment method must meet the same requirements for 
scientific validity as must be met if you use a treatment based on published literature. 
If you choose to treat seeds using a scientifically valid method to reduce microorganisms of 
public health significance at your operation (i.e., instead of relying on prior seed treatment 
applied by an entity earlier in the supply chain), you should develop a written seed treatment 
SOP as a best practice to help you improve the safety of your sprouts. The SOP should include:  
  Objectives; 
  Methods used; 
  Who is responsible for each task; 
  Materials needed; 
  Treatment procedures (e.g., how to prepare, how to monitor, how and when to discard); 
  Parameters to be measured or monitored; and  
  Relevant records to be established and kept (in compliance with the requirements of  
112.150(b)(1) and otherwise).  
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  Contains Nonbinding Recommendations 
3. Using pre-treated seeds 
There are several seed treatment methods that can be effectively applied by a grower, handler, or 
distributor of seeds such that, when followed by good handling and packaging practices, they can 
eliminate the need for you to treat seeds at your operation immediately before sprouting. 
However, you may, nonetheless, decide to re-treat pre-treated seeds. Using pre-treated seeds has 
several benefits, including increased flexibility for you and availability of treatment options that 
may be suitable for your supplier but cost-prohibitive to you (e.g., irradiation). We note that if 
you choose to purchase seeds that have been pre-treated with ionizing radiation, if you re-treat 
those same seeds with ionizing radiation, the cumulative dose must not exceed the maximum 
dose allowed by  179.26(b)(10). 
If you choose to rely, in whole or in part, on using pre-treated seeds, you must obtain 
documentation from your seed supplier that the treatment was conducted using a scientifically 
valid method to reduce microorganisms of public health significance ( 112.142(e)(2)(i)) and 
that the treated seeds were handled and packaged, following the treatment, in a manner that 
minimizes the potential for contamination ( 112.142(e)(2)(ii)). If you choose to rely solely on 
using pre-treated seeds, you must obtain documents of this type from your supplier that are 
specific to each lot of seeds you receive from that supplier. One such type of document you may 
obtain is a Certificate of Conformance. Such a certificate (or other form of documentation used 
for this purpose) should include the name or brief description of the scientifically valid method 
used to treat the seeds, the level of log reduction that the method is capable of achieving, the type 
of seeds used for any relevant validation study, and the pathogen(s) targeted by the study. We 
recommend that you also ask your seed supplier for a written explanation of the treatment 
parameters that were applied, and the basis for their conclusion that the treatment is scientifically 
valid. In the event of an investigation or inspection of your sprout operation, we may ask to 
review the science supporting the seed treatment(s) you rely on, including proprietary 
treatment(s), to ensure it is scientifically valid.   
If you intend to rely solely on a seed treatment conducted by an entity earlier in the supply chain 
(e.g., you will not re-treat the seeds yourself), you must store seeds that have received that prior 
treatment by the seed grower, distributor, or supplier under conditions that will prevent them 
from becoming contaminated (See  112.142(e)(2)(ii) and above Section V. A. 5 (Seed 
Storage)).  
4. Additional considerations for treating seeds for sprouting 
a. Water Use 
 
If you use water to prepare your seed treatment, the water is agricultural water and therefore 
must meet the microbial quality criteria in  112.44(a) (i.e., no detectable generic E. coli in 100 
mL water), and related requirements elsewhere in Subpart E. Similarly, if you rinse or soak seeds 
before, during, or after seed treatment, water used for these purposes must also meet this 
microbial quality criterion, and related requirements elsewhere in Subpart E, because such water 
contacts FCSs (surfaces of the rinse or soak container that then contact seeds used for sprouting) 
(See  112.44(a)(3)) and see Section IV (Agricultural Water in Sprout Operations)).  
38 
  Contains Nonbinding Recommendations 
b. Equipment, tools, and storage 
 
You must visually examine seeds, and packaging used to ship seeds, for signs of potential 
contamination with known or reasonably foreseeable hazards ( 112.142(d)). If you have stored 
the seeds for any length of time after their arrival at your operation and prior to sprouting, before 
bringing seeds into the treatment area, you should re-inspect bags of seeds for signs of 
contamination. Any containers or utensils that will come into contact with the seeds as part of the 
treatment process must be cleaned and sanitized before such contact ( 112.143(b)). In addition, 
if you rinse or soak seeds prior to sprouting, you must clean and sanitize the FCSs of tools and 
equipment used during this activity before they contact the seeds ( 112.143(b) and see Section 
III (Cleaning and Sanitizing)).   
c. Pre-treatment rinse, post-treatment rinse, and pre-germination seed soak 
Seeds are typically rinsed before treatment, and may also be rinsed after treatment. While the 
Produce Safety Rule does not require pre- or post-treatment rinsing, we recommend that seeds be 
rinsed thoroughly before treatment to remove dirt, and to increase the efficiency of the treatment. 
If you rinse seeds before treatment, you should repeat the rinsing process with new water until 
the visible dirt is removed and the rinse water runs clear. We also recommend that you conduct 
your rinse in such a way as to maximize seed surface contact with the water (e.g., by mixing). If 
you use a surfactant to help remove soil and debris during seed rinsing, it should be rinsed out 
completely before the next step. 
Depending on production practices and the type of seeds used, some sprout growers choose to 
use a pre-germination soak to improve germination. Soaking causes seeds to swell and softens 
their hulls to allow the sprout to grow out of the seed. Pre-germination soaking is not required by 
the Produce Safety Rule. If you rinse or soak seeds prior to sprouting, you must take any other 
measures that are reasonably necessary to prevent the introduction of known or reasonably 
foreseeable hazards into or onto the seeds during that process ( 112.142(a)). Containers used to 
rinse or soak seeds should be large enough to allow thorough mixing. We recommend that you 
change the water each time you change the seeds you are rinsing or soaking (e.g., for each new 
production batch of sprouts) to ensure that it meets the agricultural water quality standards in  
112.41 and 112.44(a). 
d. Chemical seed treatment 
If you apply a chemical seed treatment, you should use clean containers designated for mixing 
the treatment chemicals. You should take care when preparing the treatment to ensure that the 
chemical is present at the desired concentration. You should determine the quantity of treatment 
solution needed based on the quantity of the seeds to be treated. You should refer to the chemical 
label instructions to calculate the amount of chemical needed to achieve the desired 
concentration in the desired volume of treatment solution. You should use a scale to weigh dry 
chemicals before adding them to the water (or other diluent). You should stir the solution to mix 
it completely and dissolve all solids.  After mixing, you should verify the treatment solution 
concentration according to the label directions (e.g., with a test kit), since the concentration can 
impact treatment effectiveness. 
39 
  Contains Nonbinding Recommendations 
Once the chemical treatment is prepared, you should carefully combine it with the seeds. You 
should agitate the seeds and the chemical treatment at the correct temperature and for the 
appropriate amount of time according to your established procedures and the chemicals label 
instructions. It is important to use the correct amount of treatment for a known quantity of seeds; 
too much seed and/or too little chemical may decrease the effectiveness of the treatment. The 
same batch of seed treatment solution should not be used to produce more than one production 
batch of sprouts; you should prepare a new treatment solution before each production batch of 
sprouts. You should rinse seeds thoroughly after treatment to remove any residual treatment 
solution, if a rinse is recommended on the chemical label. 
C. Corrective Actions for Seeds That May Be Contaminated with a 
Pathogen 
If you learn that a lot of seeds has been associated with foodborne illness, or if you learn that a 
lot of seeds may be contaminated with a pathogen based on microbial test results (including 
those required under  112.144(b)), you are required to take certain actions with respect to that 
seed lot and sprouts grown from that seed lot ( 112.142(b)). If your seed supplier notifies you to 
stop using a lot of seed, we recommend you clarify with your supplier whether the request is due 
to potential contamination of the seed with a pathogen. You are required to maintain records 
related to corrective actions taken on seed for sprouting ( 112.150(b)(6)). You should maintain 
a record to support your decision on whether to take a corrective action on potentially 
contaminated seed, including, for example, a record of the discussion with your seed supplier, 
the reason given for asking you to not use the lot of seed, and the date on which this information 
was communicated. 
Section 112.142(b) requires that in such circumstances, except as provided in  112.142(c), you 
must discontinue use of all seeds from that lot for sprout production (112.142(b)(1)), must 
ensure that sprouts grown from that lot of seeds do not enter commerce (112.142(b)(1)), and 
must report the information to the seed grower, distributor, supplier, or other entity from whom 
the seed was purchased (112.142(b)(2)). Research indicates that seed or bean contamination, 
when it occurs, may be at low levels, intermittent, or unequally distributed within lots.  Even low 
levels of human pathogens on seeds or beans used for sprouting are a concern, due to the ideal 
growth conditions present during sprouting. Once you know or have reason to believe that a lot 
of seeds or beans is contaminated, for example through microbial testing, there is reason to 
believe that other parts of that lot may also be contaminated.  Even if you subdivide the original 
lot you receive, corrective actions are applicable to the entire original lot of seeds or beans. 
You are required to report the findings to whomever supplied the seeds to you (e.g., the seed 
grower, distributor, or supplier) so that entity can then take appropriate follow-up actions. These 
actions may include informing other buyers about the contamination of the suspected lot of 
seeds, destroying or diverting any remaining seeds from that lot to non-food uses, and/or 
investigating the potential source of contamination, as necessary. Additionally, your seed 
grower, distributor, or supplier may be required to submit a report to the Reportable Food 
Registry (RFR), which requires food facilities to report certain information to the FDA when 
there is a reasonable probability that the use of, or exposure to, an article of food will cause 
serious adverse health consequences or death to humans or animals. 
40 
  Contains Nonbinding Recommendations 
Depending on the circumstances, it may also be appropriate for you to recall sprouts that have 
already entered commerce that were produced from the affected seed lot. You should recall any 
sprouts that are adulterated. 
When your belief that a lot of seeds may be contaminated is based solely on microbial test results 
(e.g., spent sprout irrigation water test result), you would not have to take the steps described in 
112.142(b)(1) (i.e., discontinuing use of the suspect lot of seeds and ensuring that sprouts made 
from them do not enter commerce) if you treat the suspect lot with a process that is reasonably 
certain to achieve destruction or elimination of the most resistant microorganisms of public 
health significance that are likely to occur in the seeds (112.142(c)(1)). (We emphasize that 
such processes are far more robust than seed treatments described in 112.142(e), which 
typically only reduce the numbers of microorganisms of public health significance, rather than 
eliminating them.) This option exists to allow you flexibility in responding to a finding of a 
pathogen in a seed lot that would otherwise mean you would have to discontinue use of the 
seeds, and to encourage future innovation in seed treatment processes. Processes that meet the 
requirements of 112.142(c)(1) are not, at this time, commonly used in the sprouting industry.  
Further, when your belief that a lot of seeds may be contaminated is based solely on microbial 
test results (e.g., spent sprout irrigation water or in-process sprout test result), and if you 
reasonably determine, through appropriate follow-up actions, that the lot of seeds is not the 
source of contamination, you do not have to take the steps in  112.142(b)(1) or (2) (i.e., 
discontinuing use of that seed lot and ensuring that sprouts grown from that lot do not enter 
commerce), nor do you need to inform the seed grower, distributor or supplier of the positive test 
result). We acknowledge that a spent sprout irrigation water/in-process sprout sample that tests 
positive for a pathogen could be caused by contamination that originated either in the seed or 
elsewhere during the sprout growing process. Nonetheless, you must take the corrective actions 
described in  112.142(b)(1), unless you can reasonably determine, through appropriate follow-
up actions, that the lot of seeds is not the source of contamination. However, we expect that the 
situations in which you could take follow-up actions that would be adequate for you to 
reasonably determine that the lot of seeds was not the source of contamination are quite limited. 
For examples of scenarios in which we believe such a determination might be appropriate, see 80 
FR 74353 at 74500-74501 (November 27, 2015). 
Section 112.150(b)(6) requires that you establish and keep records of certain corrective actions 
including those corrective actions taken in accordance with  112.142(b) and (c) (i.e., required 
actions if you know or have reason to believe that a lot of seeds may be contaminated with a 
pathogen) and  112.148 (i.e., additional required actions where your knowledge or reason to 
believe that the lot of seeds is contaminated came from required spent sprout irrigation water or 
sprout pathogen testing pursuant to  112.144(b)). Such records must comply with all applicable 
requirements for records in Subpart O.   
In the event that your sprouts are associated with foodborne illness, you are required to 
discontinue use of all seeds from the affected lot for sprout production and ensure that sprouts 
grown from that lot of seeds do not enter commerce ( 112.142(b)(1)). Also, you must report the 
information (i.e., association of the seed lot with illness) to your grower, distributor, supplier, or 
other entity from whom you received that seed lot ( 112.142(b)(2)). We are not aware of actions 
that you could take to demonstrate that the lot of seeds was not the source of contamination 
41 
  Contains Nonbinding Recommendations 
following an outbreak of foodborne illness associated with your sprouts. Regulators may make a 
determination that the seeds were not associated with the foodborne illness outbreak, but it is 
unlikely that you would have adequate information and records (e.g., epidemiological data and 
traceback information) to make that determination independently. Therefore, in such 
circumstances you must take the actions required in  112.142(b)(1) and (2). 
To facilitate taking any necessary corrective actions associated with your seed, we recommend 
that you establish and keep sufficient records to allow you to maintain the lot identity of the 
seeds you receive and the sprouts you produce, by which we mean that you would be able to 
determine which seed lot was used to grow each production batch of sprouts, and which 
container(s) of seed is part of a lot of seed. For each seed lot, your records should allow you to 
determine the identity of the supplier, variety of the seed, date of receipt, batch or lot number of 
the seed that you received (including all identifying numbers assigned  to the seed, both by the 
supplier and by you, if different), and the date(s) that you used the  seed from that lot for 
sprouting). This information will facilitate traceback to your seed supplier and (when  possible) to 
the seed grower as needed. Maintaining records that provide traceability by connecting  each 
production batch of sprouts to the lot of seeds used to grow them can help minimize the scope of 
product potentially affected in the event of a problem with a particular production batch of 
sprouts  or a particular lot of seeds. For example, if you receive a positive test result on a spent 
sprout irrigation water test, you will need to know which seed lot(s) you used to grow that batch 
of sprouts, in order to comply with the corrective actions required in  112.142(b). If your 
records do not allow you to identify which batch of seed was used to grow the contaminated 
product, you will need to take the required corrective actions on all seed of that variety at your 
operation (e.g., if your records do not identify which lot of clover seed was used to produce a 
batch of clover sprouts that tested positive for Salmonella, you would need to discontinue use of 
all lots of clover seed at your operation in order to comply with the requirements of  
112.142(b), unless you can reasonably determine, through appropriate follow-up actions, that the 
lot of seeds is not the source of contamination). We recommend that you maintain such records 
related to each lot of seeds for at least two years after your last use of the seed lot. 
VI. Environmental Monitoring 
This section of the guidance is intended to help you comply with the requirements of the Produce 
Safety Rule for environmental monitoring of the sprout growing, harvesting, packing, and 
holding environment by sampling and testing environmental samples for Listeria species (spp.) 
or Listeria monocytogenes (L. monocytogenes). For the purposes of this guidance, environmental 
samples are samples collected from a surface or area of the growing, harvesting, packing, or 
holding environment for the purpose of testing the surface or area for the presence of Listeria 
spp. or L. monocytogenes, in accordance with the requirements of  112.144(a) and 112.145. To 
accomplish this, you must establish and implement a written environmental monitoring plan that 
is designed to identify L. monocytogenes if it is present in the growing, harvesting, packing, or 
holding environment ( 112.145(a)). As part of your environmental monitoring plan, you must 
develop a sampling plan ( 112.145(c)) that includes how often, when, and where you will 
sample the environment, and identifies the microorganism for which you will test (i.e., Listeria 
spp. or L. monocytogenes). You must collect environmental samples aseptically and test them 
using a method as set forth in  112.152 ( 112.145(d)). 
42 
  Contains Nonbinding Recommendations 
The written environmental monitoring plan must also include a corrective action plan that, at a 
minimum, requires you to take the actions in  112.146, and details when and how you will 
accomplish those actions, if the growing, harvesting, packing, or holding environment tests 
positive for Listeria spp. or L. monocytogenes ( 112.145(e)). The presence of Listeria spp. on a 
surface indicates the potential for contamination of that surface with L. monocytogenes and 
suggests that conditions are suitable for survival and/or growth of L. monocytogenes (Ref. 73).  
Section 112.146 describes the corrective actions that you must take if the growing, harvesting, 
packing, or holding environment tests positive for Listeria spp. or L. monocytogenes, which 
include: 
  Conducting additional testing of surfaces and areas surrounding the area where Listeria 
species or L. monocytogenes was detected to evaluate the extent of the problem ( 
112.146(a)) (we refer to this type of testing in this document as exploratory testing); 
  Cleaning and sanitizing the affected surfaces and surrounding areas ( 112.146(b)); 
  Conducting additional sampling and testing to determine whether the Listeria spp. or L. 
monocytogenes has been eliminated ( 112.146(c)) (we refer to this type of testing in this 
document as cleaning verification testing); 
  Conducting finished product testing when appropriate ( 112.146(d)); 
  Performing any other actions necessary to prevent recurrence of the contamination ( 
112.146(e)); and 
  Taking appropriate action to prevent any food that is adulterated under section 402 of the 
FD&C Act from entering into commerce ( 112.146(f)). 
A. Principles for Developing an Environmental Monitoring Plan 
L. monocytogenes is the target pathogenic microorganism of concern for environmental 
monitoring in a sprout operation. There are several species of Listeria, but L. monocytogenes is 
the primary species known to cause disease in humans. This pathogen is among the leading 
causes of death from foodborne illness in the United States, and predominantly affects older 
adults, pregnant women, newborns and those with weakened immune systems (Ref. 74, Ref. 75, 
Ref. 76, and Ref. 77). L. monocytogenes has been associated with foodborne illness outbreaks as 
a result of a strain of the pathogen that is transient in a food establishment.  However, more 
commonly, L. monocytogenes foodborne illness outbreaks are associated with a strain of the 
pathogen that becomes established in a food establishment (i.e., a resident strain). In such cases, 
it can result in repeated product contamination (Ref. 78). Several foodborne illness outbreaks and 
recalls involving sprouts have occurred due to contamination with L. monocytogenes, and these 
outbreaks have included fatalities (Ref. 3, Ref. 4, and Ref. 79). 
L. monocytogenes is widespread in the environment. It is found in soil, water, sewage, and 
decaying vegetation (Ref. 80, Ref. 81, Ref. 82, and Ref. 83). It can be readily isolated from 
humans, domestic animals, raw agricultural commodities, and food processing environments 
(particularly in cool, damp areas). L. monocytogenes can multiply slowly at refrigeration 
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  Contains Nonbinding Recommendations 
temperatures, thereby challenging an important defense against proliferation of foodborne 
pathogens, refrigeration (Ref. 84, Ref. 85, Ref. 86, Ref. 87, and Ref. 88). However, L. 
monocytogenes grows more quickly at warmer temperatures. While L. monocytogenes may 
occasionally be found almost anywhere in a sprout production environment, it is most likely to 
become established in areas and on surfaces that are not only wet, but are relatively undisturbed 
and that may trap organic material. These areas and surfaces include drains, cooling units, drip 
pans, areas of condensation on walls or ceilings, and areas that are difficult to access or difficult 
to clean (e.g., cracks in metal, plastic or concrete surfaces, and hollow rollers). L. monocytogenes 
is known to form biofilms (i.e., communities of microbes embedded in an organic polymer 
matrix, adhering to a surface) on food contact surfaces (FCSs) and non-FCSs and, as a result, it 
can persist on these surfaces (Ref. 89). Once L. monocytogenes has established a niche, it may 
persist in the environment for long periods of time, serving as a potential source of repeated 
contamination, until and unless the niche is identified and eliminated (Ref. 88 and Ref. 90).  
As stated above, food contact surfaces means those surfaces that contact human food and 
those surfaces from which drainage, or other transfer, onto the food or onto surfaces that contact 
the food ordinarily occurs during the normal course of operations. Food contact surfaces 
includes food contact surfaces of equipment and tools used during harvest, packing, and holding 
( 112.3). Food contact surfaces also includes FCSs of equipment and tools used in growing 
sprouts, such as those that contact seeds for sprouting (see 112.143(b)). Requirements relating to 
food contact surfaces in part 112 apply not only to surfaces that contact (or drain onto, or 
otherwise transfer onto) sprouts themselves but also those that contact seeds used to grow 
sprouts. In a sprouting operation, FCSs include, for example: trays or drums used for sprouting; 
interior surfaces of containers used for seed rinsing, seed treatment, and pre-germination seed 
soaking; and counters that come into contact with sprouts during packing and/or packaging. The 
term non-food contact surfaces refers to any surfaces that, under normal operating procedures, 
do not contact either food or FCSs, and from which drainage, or other transfer, to food or FCSs 
does not occur. Non-FCSs may include equipment, vents, fixtures, drains, walls, floors, and 
employee clothing, shoes, and accessories.  
The goals of an environmental monitoring program should be to: 
  Find L. monocytogenes and its harborage sites if present in your operation; 
  Ensure that corrective actions have eliminated L. monocytogenes and harborage sites 
when found in your operation; and 
  Determine that FCSs that are used to grow, harvest, pack or hold sprouts have been 
cleaned and sanitized prior to contact with sprouts. 
B. The Written Environmental Monitoring Plan 
The mandatory contents of your environmental monitoring plan are prescribed in  112.145, as 
discussed above. We recommend that you periodically review and assess your written 
environmental monitoring plan and update it as needed in response to new information, relevant 
changes in your operation, or corrective actions that you have taken. For example, we 
recommend updating your environmental monitoring plan when you purchase new equipment, 
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  Contains Nonbinding Recommendations 
add new product lines, make significant changes to production flow or cleaning and sanitizing 
protocols, identify new sampling locations, or identify new concerns relative to your operation as 
a result of your environmental monitoring or corrective actions taken in response to such 
monitoring. We also recommend updating your environmental monitoring plan when you 
become aware of new science relative to the control of L. monocytogenes. 
In the sections below, we describe in more detail the components of the sampling plan section of 
the environmental monitoring plan. 
C. Developing a Sampling Plan 
As part of your environmental monitoring plan, you must have a written sampling plan. Your 
written sampling plan must: 
  Specify what you will test collected samples for (i.e., Listeria species or L. 
monocytogenes) ( 112.145(c)(1)) (we refer to this microorganism as the test 
microorganism in the remainder of this document); 
  Specify the frequency of sample collection, which must be no less than monthly ( 
112.145(c)(2)); 
  Specify at what point during production you will collect the samples ( 112.145(c)(2)); 
and 
  Specify the number and location of sample collection sites, which must include 
appropriate FCS sites and non-FCS sites of equipment, and other surfaces within the 
growing, harvesting, packing, and holding environment ( 112.145(c)(3)).  
Your written sampling plan should: 
  Include a list of all FCS and non-FCS sites identified for testing at a level of detail that 
facilitates your identification of sampling sites; 
  Identify the specific test method by which collected samples will be tested for the test 
microorganism (you are required to use a method set forth in  112.152); 
  Identify the person(s) responsible for sample collection in your operation and any specific 
training that the person(s) should have (112.22(a)(3)); 
  Specify how you will meet the requirement in  112.145(d) that samples must be 
collected aseptically. Your plan should include procedures for aseptic sample collection, 
including appropriate materials and steps to prepare for sample collection; 
  Specify the sample collection method used (e.g., sponge v. swab sampling, whether any 
samples will be composited) and sample sizes to be collected at the various sample 
collection sites; 
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  Contains Nonbinding Recommendations 
  Identify the laboratory you are using to conduct the testing; and 
  Identify the records you will keep for each sample collected, including the documentation 
of the results of your analytical tests. 
D. Testing for Listeria spp. or L. monocytogenes 
Your sampling plan must specify what you will test collected samples for (i.e., the test 
microorganism, Listeria spp. or L. monocytogenes) ( 112.145(c)(1)). Testing for Listeria spp. 
will detect multiple species of Listeria, including L. monocytogenes. A positive test result for the 
presence of Listeria spp. on an FCS or non-FCS indicates the potential for contamination of that 
surface with L. monocytogenes and suggests that conditions are suitable for survival and/or 
growth of L. monocytogenes (Ref. 73). However, such a test result does not establish the 
presence of L. monocytogenes on that surface. Nonetheless, testing for either the pathogen 
directly (L. monocytogenes) or the indicator organism (Listeria spp.) facilitates accomplishing 
the objectives for environmental monitoring stated above.  If you test for Listeria spp. and take 
appropriate corrective actions when you find it on an FCS or non-FCS, you should eliminate 
Listeria spp. regardless of whether or not the initial finding of Listeria spp. was, in fact, L. 
monocytogenes. For this reason, except in certain circumstances described further below (such as 
when product testing of sprouts is appropriate as a corrective action), you are not required to take 
the added step of determining whether the identified Listeria spp. is, in fact, L. monocytogenes, 
although you may voluntarily choose to do so. Further, except in the circumstances described 
below (such as when testing for L. monocytogenes in a growing unit as a corrective action), we 
recommend that you test environmental samples for Listeria spp. because doing so will detect 
both L. monocytogenes as well as species of Listeria that are more common than L. 
monocytogenes and allow you to correct situations that could potentially lead to contamination 
with L. monocytogenes. See also the discussion in Section VI. K (Documenting and Interpreting 
Test Results) below for an explanation of how you should react to positive test results depending 
upon whether you identify Listeria spp. or L. monocytogenes as the test microorganism in your 
plan, to help you make this decision. 
When testing product (e.g., as part of corrective actions), we recommend testing for L. 
monocytogenes rather than for Listeria spp. because of the risk to public health from L. 
monocytogenes in food. If you choose to test food for Listeria spp. and find it to be positive, we 
recommend that you either determine whether the Listeria spp. is L. monocytogenes or treat the 
food as if it were contaminated with L. monocytogenes. We further recommend that a lot of 
tested product be held until the test results are received, to prevent the need for a recall, should L. 
monocytogenes be detected in the product. 
E. Person(s) Collecting Samples 
You should identify in your sampling plan the person(s) that performs sample collection, at least 
by title. Sample collection may be performed by, for example, employees or contracted 
personnel. For a larger operation, we recommend you assemble a trained Sampling Team to 
undertake this activity.  Training should be specified in your sampling plan, and your records 
should indicate that that training was successfully completed (see  112.22(a)(3)). See Section III 
(Personnel Qualification, Training, and Hygienic Practices) in the draft re-issued sections (Ref. 
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18) for additional information on training related to sample collection for environmental 
samples. Samples must be collected aseptically, in accordance with  112.145(d), and, therefore, 
training in aseptic techniques may be useful. 
F. Establishing Sample Collection Locations and Frequency 
1. Identifying sample collection locations 
You should take a risk-based approach in determining where to sample and test the environment 
for the presence of Listeria spp. or L. monocytogenes. This can be accomplished by 
characterizing the areas in your operation according to the potential for product contamination 
(e.g., using a zone system). Zone designations for surfaces or areas reflect how close those 
surfaces or areas are to a ready-to-eat food (such as sprouts), and the risk that the surfaces or 
areas pose to the food if the surfaces or areas are contaminated with L. monocytogenes (Ref. 91, 
Ref. 92, and Ref. 93). For example, you could characterize your operation with four zones as 
shown in Appendix 3 (Potential Sources of L. monocytogenes for Sampling in a Sprout 
Operation). Zone 1 is all FCSs, zone 2 is non-FCSs in close proximity to food and FCSs, zone 3 
is more remote non-FCSs that are in or near the production areas and could lead to contamination 
of zones 1 and 2, and zone 4 includes non-FCSs in remote areas outside of the production area, 
from which environmental pathogens can be introduced into the production environment. 
Establishing a zone system is not a requirement of the Produce Safety Rule, but doing so may be 
helpful in designing an environmental monitoring plan and determining the frequency for 
sampling different surfaces in different areas.  If you do not establish a zone-based system, you 
should otherwise characterize areas where you will collect environmental samples according to 
their potential for contamination or, at a minimum, distinguish between FCSs and non-FCSs. 
2. Deciding on the number of food contact surface and non-food contact surface 
sampling sites 
You must specify sample collection sites in your sampling plan, and these must include both 
FCS and non-FCS sites, which you may have designated by zone as described above ( 
112.145(c)(3)). You should make a list of FCS and non-FCS sites for potential sampling at a 
level of detail that facilitates your identification of sampling sites and sampling frequencies and 
include this list as part of your sampling plan. For examples of FCS and non-FCS sampling sites, 
see Appendix 3 (Potential Sources of L. monocytogenes for Sampling in a Sprout Operation). 
We also recommend that you assign identifiers to each of your sample sites in your sampling 
plan, particularly if your operation has more than one possible location that could meet a site 
description. For example, if you have three rotary drum growing units, you should consider 
assigning unique identifiers (e.g., Growing Unit A, Growing Unit B, Growing Unit C) to 
facilitate sample collection and so that you can associate test results with the correct sample site 
(i.e., to facilitate trend analysis and corrective actions). In addition, you may consider developing 
a diagram of your operation that identifies the FCSs and non-FCSs that you have selected as 
sampling sites. 
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  Contains Nonbinding Recommendations 
To determine the appropriate number of FCS and non-FCS sites to sample, you should consider 
the size of your operation (e.g., square footage), features of your operation (e.g. building layout), 
design of your equipment (e.g. hard to reach/disassemble/clean areas), product flow, production 
methods you use to produce the sprouts, your cleaning procedures, and any previous sampling 
results. The number of sampling sites must be sufficient to determine whether your measures are 
effective ( 112.145(c)(3)). To accomplish this requirement, your selection of sampling sites 
should be adequate to verify the implementation and effectiveness of your sanitation measures 
for controlling the presence of L. monocytogenes in the sprout production environment by 
finding Listeria spp. or L. monocytogenes if it remains in the sprouting operation after your 
routine cleaning and sanitizing procedures. We recommend you develop a full listing of FCS and 
non-FCS sampling locations that you will sample within a specified frequency, and that the plan 
rotate through different sites from that list over time such that all sites that you have identified 
for testing are tested within the time period that you have determined is appropriate (e.g., 
quarterly). If you are testing a representative subset of FCS and non-FCS sites during each 
sampling event (e.g., monthly), each surface should be assigned a priority for sampling based on 
risk.  For example, if you are using a four-zone system, we recommend that the sampling sites in 
zones 1 and 2 be sampled at a higher frequency than those in zones 3 and 4, in keeping with the 
higher risk of L. monocytogenes contamination of sprouts that zones 1 and 2 present. 
While larger sprout operations will likely have more sampling sites than smaller operations, even 
the smallest sprout growers should collect samples from at least 5 sites of FCS (that directly 
contact the food) and 5 sites of non-FCS from each production area (e.g., each growing room) 
per sampling event. Your plan should focus on those sites that are in wet production areas (such 
as growing and packing areas) that pose the greatest risk.  
As discussed in Section VI. A (Principles for Developing an Environmental Monitoring Plan), L. 
monocytogenes can be widespread in the environment, has been isolated from sprout production 
environments, and has been shown to persist on equipment and in the production environment in 
harborage sites. As a result, even if you are cleaning and sanitizing frequently, you should expect 
that you will occasionally have positive environmental samples. We discuss in subsequent 
sections of this guidance the need to take appropriate corrective actions and to reassess your 
sampling plan for possible revision to address the occurrence of positive pathogen or indicator 
organism findings. Further, if you consistently have negative test results from the sites you are 
sampling, you should reassess your sampling plan to determine whether you should add and/or 
substitute other sites to ensure that you are not missing a possible source of contamination. 
3. Identifying sampling frequency 
Your sampling plan must specify how often you will collect environmental samples, which must 
be no less than monthly. Frequency of sampling should be based on risk, which, in part, depends 
on the size and complexity of your operation. The timing of the collection of the environmental 
samples during production is an important addition to the sampling plan to ensure that sampling 
is conducted in a manner to optimize detection of Listeria, if present. While environmental 
sampling must be performed at least monthly ( 112.145(c)(2)), each sampling event need not 
include samples from all your identified sampling sites, which may be spread out over a longer 
period of time (e.g., quarterly). When developing your sampling plan, you first need to 
determine: 1) how often you will conduct environmental sampling (i.e., collect samples from 
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some of your identified sampling sites), and 2) how frequently your periodic sampling will 
ensure that you have collected samples from of all your identified sampling sites (i.e., all sites in 
your site inventory. There are various ways to achieve this, and we provide distinct examples 
below. 
a. Sampling frequency example #1 
You evaluate your operation and identify 90 sites for sampling (30 FCS, 60 non-FCS, focusing 
on non-FCSs in close proximity to FCSs), of which a representative number will be sampled 
monthly. You might determine, for example, that all FCS sites that you have identified for 
testing will be tested at least every other month and all non-FCS sites that you have identified for 
testing will be tested at least quarterly. 
In such a case you could choose to, for example, collect samples monthly, with 35 samples (from 
15 FCS and 20 non-FCS sites) at each sampling event (e.g., first production day of every month), 
as shown in Table 3. 
Table 3. Sampling Frequency Example #1 
Month  FCS Sites Sampled  Non-FCS sites Sampled 
1  Sites 1  15  Sites 1  20 
2  Sites 16  30  Sites 21  40 
3  Sites 1  15  Sites 41  60 
 
b. Sampling frequency example #2 
Like in Example 1, you again identify 90 sites for sampling (30 FCS, and 60 non-FCS). You 
might determine that both FCS and non-FCS will be sampled quarterly.  To accomplish this you  
could choose to collect a proportion of those samples every week, for a total of 30 samples 
collected over the course of each month (e.g., 5-10 samples each week), as shown in Table 4.  
Table 4. Sampling Frequency Example #2 
Month / Week    FCS Sites Sampled  Non-FCS sites Sampled 
Month 1 / Week 1   Sites 1  3  Sites 1  5 
Month 1 / Week 2   Sites 4  6  Sites 6  10 
Month 1 / Week 3   Sites 7  8  Sites 11  15 
Month 1 / Week 4   Sites 9  10  Sites 16  20 
Month 2 / Week 1   Sites 11  13  Sites 21  25 
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Month / Week    FCS Sites Sampled  Non-FCS sites Sampled 
Month 2 / Week 2   Sites 14  16  Sites 26  30 
Month 2 / Week 3   Sites 17  18  Sites 31  35 
Month 2 / Week 4   Sites 19  20  Sites 36  40 
Month 3 / Week 1   Sites 21  23  Sites 41  45 
Month 3 / Week 2   Sites 24  26  Sites 46  50 
Month 3 / Week 3   Sites 27  28  Sites 51  55 
Month 3 / Week 4   Sites 29  30  Sites 56  60 
 
You could not wait, however, for the end of each quarter to collect all 90 samples (e.g., sampling 
in January and then waiting until April to sample all 90 sites again) as this practice would not 
meet the monthly minimum sampling frequency requirement ( 112.145(c)(2)). 
c. Sampling frequency example #3 
You establish a 4-zone system that prioritizes sampling from Zones 1 and 2. There are various 
ways to achieve this. 
You might determine, for example, that all FCS sites that you identify for testing will be tested at 
least once a month and all non-FCS sites you identify for testing will be tested at least quarterly, 
recognizing that the FCS and non-FCS sites that you identify for testing are representative of the 
total number of FCS and non-FCS that you have identified in your operation. 
In such a case you could choose to, for example, specify sample collection from specific FCS 
sites at least once every week, such that all FCS sites that you identify for testing are tested at 
least once each month. 
As part of such an approach, you might choose to, for example, specify sample collection from 
representative sets of Zone 2 non-FCS sites every two weeks and sample collection from Zone 3 
and 4 sites monthly, such that all non-FCS sites that you identify for testing are tested at least 
once each quarter. 
G. Timing of Sample Collection 
Your sampling plan must specify the point(s) during production at which you will collect 
environmental samples ( 112.145(c)(2)). We recommend you collect environmental samples 
several hours after the start of production (e.g., 3 to 4 hours), preferably towards the end of 
production, just prior to cleanup. Collecting your samples toward the end of production allows L. 
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  Contains Nonbinding Recommendations 
monocytogenes (if present) to work its way out of harborage sites and into the environment 
where it can more easily be detected. 
Environmental samples should not be taken immediately after surfaces have been sanitized, as 
the sanitizer may affect the test results. The timing of collection of environmental samples for 
identifying L. monocytogenes should not be confused with that of other types of samples 
collected for verification of cleaning and sanitizing, which are typically collected immediately 
after sanitizing. For example, ATP hygiene monitoring involves the use of a device called a 
luminometer to measure the combined total ATP of organic material (food residues and 
microbial populations) collected from a swabbed surface. See discussion on verification of 
cleaning and sanitizing in Section III (Cleaning and Sanitizing) of this guidance for more 
information. 
H. Sample Collection and Shipping 
While not required by the Produce Safety Rule, we recommend your sampling plan specify the 
sampling method that you will use and detail your sample collection procedures. Use of 
appropriate materials and equipment is particularly important to ensure that sample collection is 
conducted aseptically, as required by  112.145(d). For more details on aseptic sampling and 
information on sampling methods and sample collection, see Appendix 1 (Aseptic Sampling) and 
Appendix 2 (Collection Procedures  Environmental Sampling). 
Prior to and during delivery or shipment to a laboratory, samples should be kept refrigerated. 
Sealed coolant packs should be used in lieu of ice, as needed during delivery or shipment, to 
avoid the possibility of melting ice contaminating the sample. Samples should not be frozen. 
Samples should be shipped so that they are received by the laboratory within 24 hours after 
sample collection. We recommend that the maximum timeframe between environmental 
sampling and analysis be 48 hours (Ref. 94).  
Prior to sending the samples to a laboratory for testing, you should verify that your samples are 
clearly identified with the date and location of collection of the samples and the name of your 
sprouting operation. You should also specify the microorganism for testing. 
I. Choosing a Laboratory  
You should choose a laboratory that is qualified to test environmental samples for Listeria spp. 
or L. monocytogenes (whichever is your test microorganism). Testing is typically contracted to a 
third-party testing laboratory, but may also be performed by your own laboratory (e.g., your own 
in-house laboratory). If you use an in-house laboratory, you may be able to complete the 
screening step of the test method, but may need to send the enrichment broth out to a third-party 
testing laboratory for any confirmatory testing (if it cannot be conducted in-house). If using a 
third-party testing laboratory, you should use a laboratory that employs scientifically valid 
laboratory methods and procedures that can provide reliable, accurate test results. Using an 
accredited laboratory (e.g., a laboratory accredited to International Organization for 
Standardization (ISO)/IEC 17025:2017, General requirements for the competence of testing and 
calibration laboratories, Third edition, November 2017) is one way to have confidence that the 
laboratory will provide reliable, accurate test results. Regardless of which laboratory you use, 
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  Contains Nonbinding Recommendations 
testing must be done using a method as set forth in  112.152. You should ask the laboratory 
which test methods will be used when analyzing environmental samples for Listeria spp. or L. 
monocytogenes. We recommend you use a laboratory that either utilizes the method listed in  
112.152(a) or an alternate method reviewed by FDA and found to be equivalent that has been 
made available on our website (available at https://www.fda.gov/food/laboratory-methods-
food/equivalent-testing-methodologies-listeria-species-and-listeria-monocytogenes-
environmental-samples).  If the laboratory test methods do not meet the requirements of  
112.152, you may consider selecting another laboratory to analyze your samples or request that 
the laboratory utilize a different method.  
In addition, FDA published a final rule to establish the laboratory accreditation for analyses of 
foods (LAAF) program as required by FSMA. This final rule outlines the procedures and 
standards that accreditation bodies and laboratories will need to follow to participate in the 
LAAF program, as well as procedures for how FDA will manage and oversee the LAAF 
program. Once fully implemented, the final rule will require sprout operations subject to Subpart 
M to use a LAAF-accredited laboratory for corrective action testing of environmental samples ( 
112.146(a) and (c)) and finished product ( 112.146(d)). See 21 CFR 1.1107(a)(1)(i). 
J. Choosing a Test Method for Listeria spp. or L. monocytogenes 
In accordance with  112.152, the growing, harvesting, packing, and holding environment must 
be tested for Listeria spp. or L. monocytogenes using Testing Methodology for Listeria species 
or L. monocytogenes in Environmental Samples (currently available at  
https://www.fda.gov/media/94358/download6 ( 112.152(a))) or a scientifically valid method 
that is at least equivalent to FDAs method in accuracy, precision, and sensitivity ( 112.152(b)). 
If your laboratory utilizes a different method to test your environmental samples, we recommend 
you use the questions included in the decision tree for test methods requirements for Salmonella 
and E. coli O157:H7 in spent sprout irrigation water (or sprouts) (available at 
https://www.fda.gov/media/132369/download) when you are determining whether the method 
may be equivalent to the method listed in  112.152(a).  
If you test sprouts for Listeria monocytogenes as part of a corrective action, we recommend that 
you use the procedures described in FDAs Bacteriological Analytical Manual Online (BAM), 
Chapter 10  Listeria monocytogenes, Detection and Enumeration of Listeria monocytogenes 
in Foods (currently available at: https://www.fda.gov/food/laboratory-methods-food/bam-
chapter-10-detection-listeria-monocytogenes-foods-and-environmental-samples-and-
enumeration).6  
If you plan to use a test method other than the one listed in  112.152(a) to test the growing, 
harvesting, packing, and holding environment for Listeria spp. or L. monocytogenes, it must be a 
scientifically valid method that is at least equivalent to the method in  112.152(a) in accuracy, 
 
6 Because websites are subject to change, it is possible that this specific website address will change. If you cannot 
access this document at that website, alternative websites where you currently can access this document include  
https://www.fda.gov/food/science-research-food/laboratory-methods-food, and https://www.fda.gov/food/guidance-
regulation-food-and-dietary-supplements/food-safety-modernization-act-fsma. Alternatively, you can search on part 
or all of the title of the method, in the search box on FDAs Web site at https://www.fda.gov/ or using a generally 
available search engine. 
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  Contains Nonbinding Recommendations 
precision, and sensitivity ( 112.152(b)). We use the term scientifically valid to mean an 
approach that is based on scientific information, data, or results published in, for example, 
scientific journals, references, textbooks, or proprietary research. Although you are not required 
to notify or submit information to FDA prior to using such an alternate method, you must 
establish and keep records of any such alternate methods that you use ( 112.150(b)(5)). For 
example, your records could specify that you are using one of the methods that FDA listed in 
Equivalent Testing Methodologies for Listeria species and Listeria monocytogenes in 
Environmental Samples on our website (available at https://www.fda.gov/food/laboratory-
methods-food/equivalent-testing-methodologies-listeria-species-and-listeria-monocytogenes-
environmental-samples). If the alternate method that you use is not listed by FDA, your records 
regarding alternate methods should include the 1) name and identification number of the method 
validated by a third-party methods validation organization, or 2) detailed analytical procedures, 
an explanation of why the alternate method is at least equivalent to the reference method (such as 
a scientific study, or data or information regarding the sensitivity, specificity, and precision of 
the alternate method compared to the reference method), and any other relevant information 
supporting the use of the alternate method.   
As noted in the list of Equivalent Testing Methodologies for Listeria species and Listeria 
monocytogenes in Environmental Samples, FDAs method in the BAM for testing Listeria spp. 
and L. monocytogenes in environmental samples (available at 
https://www.fda.gov/food/laboratory-methods-food/bam-chapter-10-detection-listeria-
monocytogenes-foods-and-environmental-samples-and-enumeration) is another method that is 
considered to be at least equivalent to the method in  112.152(a) in accuracy, precision, and 
sensitivity and can be used as an alternate test method (as permitted in  112.152(b)).  
We recommend use of methods validated through a collaborative study (such methods are 
currently available at https://www.fda.gov/media/83812/download)7, for example per AOAC 
Appendix J or ISO 16140:2016. Alternate methods should be compared to FDAs reference 
method ( 112.152(a)) in environmental samples and demonstrated to meet the requirements for 
alternate methods in  112.152(b) (i.e., demonstrated to be at least equivalent to the reference 
method in accuracy, precision, and sensitivity). Methods validated by third-party methods 
validation organizations such as AOAC Official Methods of Analysis (OMA), MicroVal, and 
Association Francaise de Normalisation (AFNOR) may meet the requirements in  112.152(b); 
however, FDA does not automatically consider methods validated by third-party organizations 
such as the listed organizations to be equivalent to the reference method. Information on 
alternate methods reviewed by FDA and found to be equivalent will be made available on our 
website, such as at https://www.fda.gov/food/guidance-regulation-food-and-dietary-
supplements/food-safety-modernization-act-fsma and https://www.fda.gov/food/science-
research-food/laboratory-methods-food.  
 
 
7 Alternative websites where you can currently access this document include: 
 https://www.fda.gov/science-research/field-science-and-laboratories/method-validation-guidelines and 
https://www.fda.gov/food/guidance-regulation-food-and-dietary-supplements/food-safety-modernization-act-fsma. 
Alternatively, you can search on part or all of the title of the method, in the search box on FDAs Web site at 
https://www.fda.gov/ or using a generally available search engine. 
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K. Documenting and Interpreting Test Results 
Section 112.150(b)(4) requires that you establish and keep records of test results from 
environmental testing for Listeria spp. or L. monocytogenes you perform in compliance with  
112.144(a) and 112.145 or any additional testing conducted if you detect Listeria spp. or L. 
monocytogenes in the environment in compliance with  112.146 (a), (c), (d), and (e). The results 
of all analytical tests must be documented, regardless of whether they were conducted by your 
own (e.g., in-house) laboratories or third-party laboratories. 
Testing for Listeria spp. or L. monocytogenes in environmental samples using the FDA reference 
method ( 112.152(a)) can yield one of the following results: 
  A positive result for Listeria spp., which for purposes of the FDA reference method, 
means finding the presence of typical colonies on a Listeria specific agar (i.e., completing 
Steps I.A-I.F in the FDA reference method with respect to Listeria).8 
  A positive result for L. monocytogenes, which can be obtained if a positive result for 
Listeria spp., or a presumptive positive result for L. monocytogenes, is followed by 
confirmatory steps that result in a confirmed L. monocytogenes cultural isolate (i.e., 
completing Step I.I in the FDA reference method). 
  One of two types of negative results: 
  Listeria specific agars do not yield a positive result for Listeria spp., indicating a 
o
negative finding for Listeria spp.; or 
  Additional confirmatory steps after a positive result for Listeria spp. or a presumptive 
o
positive result for L. monocytogenes do not result in confirmation of the presence of 
L. monocytogenes, indicating that while Listeria spp. were found, L. monocytogenes 
was not found. 
1. Sampling plan that specifies testing for Listeria spp. (recommended approach) 
If you choose to establish a sampling plan that specifies testing for Listeria spp., you are not 
required to perform confirmatory testing for L. monocytogenes after obtaining a positive test 
result for Listeria spp., although you may voluntarily choose to do so. 
If your plan specifies that you will test for Listeria spp. and you conduct testing using the FDA 
reference method (recommended approach): 
 
8 The FDA reference method provides options for testing for both Listeria spp. and L. monocytogenes. They are 
described in the reference method in parallel, such that, for example, step I.E.1 describes part of the procedure for 
testing for L. monocytogenes while step I.E.2 describes part of the procedure for testing for Listeria spp. Thus, a 
positive result for Listeria spp. refers to completion of the reference method up to step I.F (or G, if applicable) 
including all steps relevant to Listeria spp. 
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  A positive result for Listeria spp. triggers the requirement to take corrective actions under 
 112.146; 
  A negative result for Listeria spp. does not trigger the requirement to take corrective 
actions under  112.146; 
  If you voluntarily choose to perform confirmatory testing for L. monocytogenes on an 
environmental sample that yields a positive result for Listeria spp., a positive result for L. 
monocytogenes also triggers the requirement to take corrective actions under  112.146; 
and 
  If you voluntarily choose to perform confirmatory testing for L. monocytogenes on an 
environmental sample that yields a positive result for Listeria spp., a negative result for 
L. monocytogenes does not eliminate the requirement to take corrective actions under  
112.146 based on your positive result for Listeria spp. A positive test result for the 
presence of Listeria spp. on an FCS or non-FCS indicates the potential for contamination 
of that surface with L. monocytogenes and suggests that conditions are suitable for 
survival and/or growth of L. monocytogenes (Ref. 73). Therefore, if your tests identify 
the presence of Listeria spp., you should eliminate Listeria spp., regardless of whether or 
not the initial finding of Listeria spp. was, in fact, L. monocytogenes.   
2. Sampling plan that includes testing for L. monocytogenes 
If you choose to establish a plan that specifies that you will test for L. monocytogenes and you 
conduct testing using the FDA reference method: 
  A positive result for Listeria spp. triggers the requirement to take corrective actions under 
 112.146, and you must continue with confirmatory testing to identify L. monocytogenes 
as provided in your sampling plan; 
  A negative result for Listeria spp. does not trigger the requirement to take corrective 
actions under  112.146; 
  A positive result for L. monocytogenes triggers the requirement to take corrective actions 
under  112.146; and 
  A negative result for L. monocytogenes does not eliminate the requirement to take 
corrective actions under  112.146 based on your positive result for Listeria spp. A 
positive test result for the presence of Listeria spp. on an FCS or non-FCS indicates the 
potential for contamination of that surface with L. monocytogenes and suggests that 
conditions are suitable for survival and/or growth of L. monocytogenes (Ref. 73). 
Therefore, if you identify Listeria spp., you should eliminate Listeria spp., regardless of 
whether or not the initial finding of Listeria spp. was, in fact, L. monocytogenes.   
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L. Developing a Corrective Action Plan and Taking Corrective Actions 
Your written environmental monitoring plan must also include a corrective action plan that, at a 
minimum, requires you to take the actions in  112.146, and details when and how you will 
accomplish those actions, if the growing, harvesting, packing, or holding environment tests 
positive for Listeria spp. or L. monocytogenes ( 112.145(e)). 
1. Corrective action plan 
Having a corrective action plan in place at your operation will help ensure that you take 
corrective actions quickly and consistently in response to a finding of Listeria spp. or L. 
monocytogenes in the environment. Your corrective action plan must ( 112.145(e)): 
  Specify when and how you will conduct additional testing of surfaces and areas 
surrounding the area where the positive test result was detected to evaluate the extent of 
the problem, including the potential for Listeria spp. or L. monocytogenes to have 
become established in a niche ( 112.146(a)) (i.e., exploratory testing); 
  Specify how and when you will clean and sanitize the affected surfaces and surrounding 
areas ( 112.146(b)); 
  Specify when and how you will conduct additional sampling and testing to determine 
whether the Listeria spp. or L. monocytogenes has been eliminated ( 112.146(c)) (i.e., 
cleaning verification testing); 
  Specify when and how you will conduct finished product testing when appropriate ( 
112.146(d)); 
  Indicate that you will perform any other actions necessary to prevent recurrence of the 
contamination ( 112.146(e)) and should specify what those actions may be.  For 
example, you should specify additional steps you will take to determine the source and 
route of contamination if your exploratory testing or cleaning verification testing yields 
positive results for Listeria spp. or L. monocytogenes and steps you will take to eliminate 
such sources and routes of contamination; and 
  Specify when and how you will take appropriate action to prevent any food that is 
adulterated under section 402 of the FD&C Act from entering into commerce ( 
112.146(f)). 
2. Implementing corrective actions 
We recommend that you consider what corrective action is appropriate in a specific set of 
circumstances based, in part, on whether or not you detected Listeria spp. or L. monocytogenes, 
whether or not you detected the organism on an FCS or a non-FCS site, and whether or not this 
was the first time you detected the organism on the surface. Based on the criteria described 
above, Table 5 provides a key to the corrective actions described in the sections below. 
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Table 5. Corrective Actions When Listeria Species or L. monocytogenes is Found in an 
Environmental Sample 
Test Microorganism  Type of Surface or Product  Positive*  Sprout 
where Positive was Found  Guidance 
Section 
Listeria spp.  FCS  1st positive  L.3.a.  
Listeria spp.  FCS  2nd positive  L.3.b. 
Listeria spp.  FCS  3rd or subsequent  L.3.c. 
positive 
L. monocytogenes  Product  Any  L.3.c. 
Listeria spp.  FCS in a growing unit  Any  L.3.d. 
L. monocytogenes  FCS or non-FCS  Any  L.4. 
Listeria spp.  Non-FCS  1st positive  L.5.a.  
Listeria spp.  Non-FCS  2nd positive  L.5.b. 
Listeria spp.  Non-FCS  3rd or subsequent  L.5.c. 
positive 
*For the purposes of this table, 2nd, 3rd, etc. positives refer to positives identified during 
corrective actions in response to the 1st positive identified through routine monitoring. 
3. Corrective actions if you detect Listeria spp. on a food contact surface 
We describe appropriate corrective actions for a positive test result for Listeria spp. from an 
environmental sample collected during your routine sampling of an FCS site in this section. 
These steps are also summarized in Figure 3 (FCS* Testing and Follow-Up Activities Decision 
Tree). 
a. Listeria spp. positive on an FCS (1st positive) 
You should examine the area surrounding the site of the positive test result in all directions for 
potential sources of Listeria spp. as described in Appendix 3 (Potential Sources of L. 
monocytogenes for Sampling in a Sprout Operation) of this guidance. You should pay particular 
attention to possible niches that provide harborage for L. monocytogenes. 
i.  Exploratory testing 
You must conduct additional testing of surfaces and areas surrounding the area where Listeria 
spp. was detected to evaluate the extent of the problem, including the potential for Listeria spp. 
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or L. monocytogenes to have become established in a niche ( 112.146(a)). Consider the 
following: 
  Exploratory testing provides you with information about the extent of the problem 
represented by the initial positive test result (e.g., whether the presence of Listeria spp. is 
isolated or more extensive). You do not need to wait for the results of the exploratory 
testing before doing any cleaning and sanitizing, but you should use the results of 
exploratory testing to inform your decision about the extent of cleaning and sanitizing 
that you will perform, as required by  112.146(b) (see next subheading);   
  You should conduct exploratory sampling and testing of sites that represent a potential 
source of contamination to the contaminated FCS site. Exploratory testing should include 
at least 3 samples from sites in close proximity to the positive site and those upstream in 
the operation (i.e., locations in the operation that the product contacts earlier in the 
product flow) to help identify the source of contamination; 
  Exploratory testing while in production: If you receive a positive Listeria spp. result for a 
routine sample from an FCS site while you are in production (e.g., you are either still 
growing the production batch of sprouts that was growing when you took your routine 
sample, or you have started another production batch of sprouts), you should conduct 
exploratory testing during that production cycle;  
  Exploratory testing when not in production: If you receive a positive result for a routine 
sample from an FCS site when you are not in production (e.g., the production batch of 
sprouts that was growing when you took your routine sample has been completed and 
you have not started the next production batch), you should conduct exploratory testing 
during the production of the next batch; and 
  If you receive any positive results from your exploratory testing, you should conduct 
intensified cleaning and sanitizing and intensified sampling and testing (discussed below 
 
as a recommended response to a 2ndpositive). 
ii.  Cleaning and sanitizing 
You must clean and sanitize the affected surfaces, including FCSs and non-FCSs, as well as 
surfaces from the surrounding area where the positive sample was taken ( 112.146(b); see also 
 112.123(d)(1)-(2); 112.143(b)). Consider the following: 
  At a minimum, the site of the initial positive Listeria spp. result and the sites of any 
additional positive results found during exploratory testing should all be cleaned and 
sanitized before your next production run; and 
  You should also consider verifying the efficacy of your cleaning and sanitizing using 
additional methods (e.g., ATP testing) beyond the required testing ( 112.146(c)) before 
your next production run; see Section III (Cleaning and Sanitizing), and 
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  If you receive negative test results from all of your exploratory testing, and you have 
cleaned and sanitized the affected FCSs and non-FCSs from the surrounding area where 
the original positive sample was taken, you may consider the actions required by 
112.146(b) and 112.146(c) to have been accomplished while conducting the actions 
required by  112.146(a). 
b. Listeria spp. positive on an FCS during exploratory testing (2nd positive)  
i.  Three production days of intensified cleaning and sanitizing 
If any of the exploratory testing samples are positive for Listeria spp., as an action to prevent 
recurrence of the contamination, you must clean and sanitize the affected surfaces and 
surrounding areas ( 112.146(b)). The cleaning and sanitizing that you perform under these 
circumstances should be intensified beyond your routine efforts and should be performed in the 
affected areas for the next three production days. Intensified cleaning and sanitizing include 
sanitation measures that you perform in addition to your normal sanitation procedures. See also 
Section III.F. (Cleaning and Sanitizing Conducted in Response to Suspected or Known 
Contamination). You should escalate the intensity of these measures in response to additional 
positive Listeria spp. samples. Intensified cleaning and sanitizing can include: 
  Increasing the frequency of cleaning and sanitizing for certain pieces of equipment;  
  Breaking down the equipment into its parts for further cleaning;  
  The use of more effective or different cleaning and sanitizing methods or agents (e.g. 
more aggressive scrubbing, applying heat (where possible)) (see Section III (Cleaning 
and Sanitizing)); 
  Intensified (e.g., more vigorous, longer duration) scrubbing of surfaces; 
  Identifying, cleaning, and sanitizing of possible harborage sites and possible cross- 
contamination routes; and 
  Soaking of equipment parts in an appropriate sanitizing agent for an appropriate duration. 
ii.  Three production days of intensified testing 
If any of the exploratory testing samples are positive for Listeria spp., as an action to prevent 
recurrence of the contamination ( 112.146(c) and 112.146(e)), you should conduct a round of 
intensified testing during each of the next three production days, both to verify the effectiveness 
of your intensified cleaning and sanitizing, and to look for possible harborage sites in the 
affected area. To look for possible harborage sites, you should disassemble equipment as 
appropriate, and sample and test areas of the equipment exposed by disassembly prior to 
cleaning and sanitizing the equipment. Each round of intensified testing should include at least 3 
samples, including the initial Listeria spp. positive site, surrounding positive sites that tested 
positive for Listeria spp. exploratory testing, and surrounding FCS and non-FCS sites in close 
proximity to any positive sites.  
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iii.  Finished product testing and other product actions 
You must conduct finished product testing when appropriate ( 112.146(d)). In the 
circumstances described here, where you have identified a second positive result on an FCS, you 
should: 
  Test the production batch(es) of sprouts from the production day associated with the 
second positive for Listeria spp. on the FCS. You should test the sprouts for L. 
monocytogenes using a statistically-based sampling protocol and analytical methods that 
will provide an appropriate level of confidence in the results (e.g., 95% confidence that 
you will detect L. monocytogenes in the sample if it is present). The International 
Commission on Microbiological Specifications for Foods (ICMSF) provides additional 
information on scientifically-based sampling plans that can be used to provide statistical 
confidence for results of product testing (Ref. 95). While these test results are pending, 
and while you are taking the other steps recommended in this section (i.e., three 
production days of intensified cleaning and sanitizing, three production days of 
intensified sampling and testing, receiving results from such testing, conducting a 
comprehensive investigation), you should not allow the production batch(es) of sprouts to 
enter commerce. 
iv.  Comprehensive investigation 
Following a second positive finding of Listeria spp. on an FCS, as an action to prevent 
recurrence of the contamination ( 112.146(e)), you should conduct a comprehensive 
investigation to identify and address Listeria sources, and modify your sanitation procedures, 
where appropriate. You may need to stop production at your sprout operation in order to conduct 
the comprehensive investigation or to take actions based on the comprehensive investigation 
(e.g., repair or replace equipment). Such an investigation should include: 
  Checking maintenance records for modifications or repairs to relevant equipment; 
  Interviewing and observing sanitation, maintenance, and production employees to 
determine whether appropriate procedures are being followed, including hygienic 
practices; 
  Reviewing production, maintenance, and sanitation procedures to determine whether 
modification of the procedures is necessary to prevent contamination; and 
  Reviewing traffic patterns and equipment layout. 
Based on the results of the comprehensive investigation, you should correct any identified 
problems (e.g., re-train personnel, revise sanitation procedures, repair equipment, update 
maintenance program) to prevent recurrence of contamination ( 112.21(d) and 112.146(e)). 
v.  Negative results from intensified testing and finished product testing 
If all results from your three production days of intensified testing are negative, and your 
finished product testing is also negative, you should return to routine environmental monitoring. 
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It would be reasonable to allow all sprouts from all three production days to enter commerce at 
this point, provided there is no other reason for concern (e.g., all other testing requirements in  
112.147 have been satisfied for these batches). However, you should target those surfaces where 
positive test results were found for sampling and testing during your next routine environmental 
sampling event. 
c. Listeria spp. positive during intensified testing (3rd or subsequent positive) or L. 
monocytogenes positive product 
i.  Stop production and investigate 
If your intensified sampling and testing again detects Listeria spp. on an FCS and/or a non-FCS 
site (third or subsequent positive), and/or you detect L. monocytogenes in your product, you 
should assume that you have a harborage site.  As an action to prevent recurrence of the 
contamination ( 112.146(e)), you should stop production and consult a food safety expert 
familiar with troubleshooting L. monocytogenes contamination problems to conduct a 
comprehensive investigation and make recommendations for appropriate actions for you to take 
based upon that investigation. You could choose to seek more information from sources such as 
cleaning and sanitation suppliers, equipment manufacturers, academia, extension services, and 
industry associations to assist you with these corrective action steps. 
ii.  Product actions 
You also must take appropriate action to prevent any food that is adulterated under section 402 
of the FD&C Act from entering into commerce ( 112.146(f)). If a production batch of sprouts 
tests positive for L. monocytogenes,  112.146(f) requires you to prevent it from entering into 
commerce. We recommend that you destroy any such production batch. If you are following the 
three production days of intensified testing described above in L.3.b, you also should consider 
the production batches during all three days of intensified testing to be adulterated, considering 
the positive finding of L. monocytogenes in the first batch, combined with the earlier positive 
findings in your environment. Moreover, we recommend that you destroy any such batches if 
you have held them or recall them if you have already shipped them. You should also evaluate 
whether any other production batches of sprouts (either at your operation or in distribution) 
should be recalled or destroyed. 
iii.  Returning to production 
After all of these corrective actions have been taken and production begins again, you should 
prevent your new production batches of sprouts from entering commerce until you test each 
production batch of sprouts for L. monocytogenes and conduct intensified testing of the 
environment on each production day, until you have received three consecutive days of negative 
test results for FCSs, non-FCSs, and sprouts. After receiving these negative test results for three 
consecutive days, you would not need to continue to test each production batch of sprouts for L. 
monocytogenes. 
The example in Figure 3 addresses testing and follow-up actions for a single positive finding of 
Listeria spp. on an FCS during one sampling period. Detecting Listeria spp. at several FCS 
sampling locations during the same sampling period could indicate that your routine sanitation 
61 
  Contains Nonbinding Recommendations 
procedures are inadequate and could indicate that Listeria spp. has more broadly established in 
your operation. The risk associated with cross-contamination from contaminated FCS sites to 
food increases as the number of contaminated FCS sites increases. Therefore, when more than 
one FCS site sample tests positive for Listeria spp. during one sampling period, you should 
immediately review your written cleaning and sanitation procedures to identify and implement 
more effective measures and escalate your corrective actions until you have identified and 
eliminated the Listeria spp. source(s). 
Figure 3. FCS* Testing and Follow-Up Activities Decision Tree 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
   
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d. Additional considerations if you detect Listeria spp. on an FCS in a growing unit 
(e.g., rotary drum growing units) 
If you obtain a positive for Listeria spp. from an FCS in a growing unit (e.g., FCS on a rotary 
drum growing unit) during growing, there is a heightened risk that your sprouts may be 
contaminated with L. monocytogenes. As a result, we recommend that you take additional 
corrective actions in response to the finding, beyond those already discussed above. 
If you receive notification of a positive test result for Listeria spp. from an FCS in a growing unit 
collected during your routine sampling (i.e., a first environmental positive from such a site), you 
should take all of the corrective actions described above relative to such a finding and should 
also take action to prevent the production batch(es) of sprouts associated with the positive 
sample site (i.e., the production batch of sprouts that was grown in that growing unit in which the 
positive Listeria spp. sample was collected at the time it was collected and any other potentially 
affected product) from entering commerce while you take the following steps: 
1)  Conduct exploratory testing as described above ( 112.146(a)). If your exploratory testing 
yields additional positive samples for Listeria spp., you should conduct intensified cleaning 
and sanitizing, and intensified testing, for three consecutive production days (as described 
 
above as a recommended response to a 2ndListeria spp. positive on an FCS site). If any of 
your intensified testing yields a positive result, you should proceed to the corrective actions 
recommended above for a 3rd Listeria spp. positive on an FCS site. 
2)  Further analyze the sample that was positive for Listeria spp. to determine whether the 
Listeria identified is L. monocytogenes. If you determine the sample is positive for L. 
monocytogenes,  112.146(f) requires you to take appropriate action to prevent the 
production batch of sprouts grown in the affected growing unit from entering commerce.  We 
recommend that you destroy any such product. 
3)  If all of your intensified testing of the environment for Listeria spp. for three consecutive 
production days, and L. monocytogenes finished product testing is negative, it would be 
reasonable at that time to allow the production batch of sprouts at issue to enter commerce, 
and to return to routine sampling and testing. 
4. Corrective actions if you detect Listeria monocytogenes on a food contact surface or 
non-food contact surface 
In general, we anticipate that sprout operations will elect to test FCS and non-FCS sites for 
Listeria spp., rather than L. monocytogenes. Ordinarily, there is minimal value in determining 
whether Listeria spp. is L. monocytogenes because, typically, you should focus on eliminating 
the Listeria spp. regardless of whether it is, in fact, L. monocytogenes. However, in certain cases 
you should consider conducting further tests to determine whether the Listeria spp. positive in 
your environmental samples is L. monocytogenes. One example of such a situation is described 
above in Section VI. L. 3. d (Additional considerations if you detect Listeria spp. on an FCS in a 
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growing unit (e.g., rotary drum growing units)) for positive Listeria spp. findings on an FCS in a 
growing unit. 
If you detect L. monocytogenes on an FCS,  112.146(f) requires to you take appropriate action 
to prevent any food that is adulterated under section 402 of the FD&C Act from entering into 
commerce. Depending on the circumstances, you may have production batches of sprouts that 
are adulterated under section 402(a)(4) of the FD&C Act because of their association with the 
affected FCS location. We recommend that you destroy any potentially affected production batch 
of sprouts (or other food) associated with the contaminated FCS (as part of a recall, if applicable) 
and follow procedures outlined above in the subsection titled returning to production in 
Section VI. L. 3. c. (Listeria spp. positive during intensified testing (3rd or subsequent positive) 
or L. monocytogenes positive product).   
Section 112.150(b)(6) requires that you establish and keep records of corrective actions 
conducted in accordance with the requirements of  112.146. Such records must include: 
  Documentation of additional testing of surfaces and areas surrounding the area where the 
positive test result was detected to evaluate the extent of the problem, including the 
potential for Listeria spp. or L. monocytogenes to have become established in a niche ( 
112.146(a)) (i.e., exploratory testing); 
  Documentation of cleaning and sanitizing the affected surfaces and surrounding  areas ( 
112.146(b)); 
  Documentation of additional sampling and testing to determine whether the Listeria spp. 
or L. monocytogenes has been eliminated ( 112.146(c)); 
  Documentation of finished product testing when appropriate (  112.146(d)); 
  Documentation of any other actions necessary to prevent recurrence of the  contamination 
( 112.146(e)); and 
  Documentation of appropriate action to prevent any food that is adulterated under section 
402 of the FD&C Act from entering into commerce ( 112.146(f)). 
5. Corrective actions if you detect Listeria spp. on a non-food contact surface 
We describe appropriate corrective actions for a positive test result for Listeria spp. from an 
environmental sample collected during your routine sampling of a non-FCS site in this section. 
These steps are also summarized in Figure 4 (Non-FCS* Testing and Follow-Up Activities for 
Zone 2 Decision Tree). We focus on corrective actions for positives in Zone 2, which are in close 
proximity to food and FCS, because, when contaminated, they present a greater risk of 
contamination of the food than do non-FCS in other zones. However, as required by  112.146, 
you must also take corrective actions if a positive result(s) is obtained in Zones 3 or 4. Corrective 
actions for non-FCS positives in Zones 3 and 4 may be less rigorous than those for non-FCS 
positives in Zone 2 provided that all relevant requirements are met. For example, you must 
conduct additional testing (i.e., exploratory testing, see Section VI. L. 3. a (Listeria spp. 
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positive on an FCS (1st positive))) of surfaces and areas surrounding the area where Listeria 
species or L. monocytogenes was detected ( 112.146(a)). However, it would be reasonable to 
take fewer exploratory samples after a positive test result in Zone 3 or 4 compared to the number 
of exploratory samples taken after a positive test result in Zone 2. You must clean and sanitize 
the affected surfaces and surrounding areas after obtaining a positive test result in Zone 3 or 4 ( 
112.146(b)), however the cleaning and sanitizing conducted might be more aggressive after a 
positive test result in Zone 2 compared to cleaning and sanitizing after a positive test result in 
Zone 3 or 4. You must then conduct additional sampling and testing (see Section VI. L. 3. a 
(Listeria spp. positive on an FCS (1st positive)) to determine whether the Listeria species or L. 
monocytogenes has been eliminated after obtaining a positive test result in Zone 3 or 4 ( 
112.146(c)). However, it would be reasonable to take more samples as a follow-up for a positive 
test result in Zone 2 compared to the number of samples taken after a positive test result in Zone 
3 or 4. Finished product testing is also more likely to be necessary in response to a positive test 
result in Zone 2 compared to after a positive test result in Zone 3 or 4.  
The discussion in this section relates only to positive environmental samples on non-FCSs. If 
you obtain a positive environmental sample (e.g., Listeria spp.) on an FCS during follow-up 
testing to an original positive on a non-FCS (e.g., during exploratory testing), you should 
immediately switch to taking corrective actions appropriate to positive findings on FCSs, which 
is covered in Section VI. L. 3. b (Listeria spp. positive on FCS during exploratory testing (2nd 
positive)) above. 
a. Listeria spp. positive on non-FCS (1st positive) 
You should examine the area surrounding the site of the positive test result in all directions for 
potential sources of Listeria spp. as described in Appendix 3 (Potential Sources of L. 
monocytogenes for Sampling in a Sprout Operation) of this guidance. You should pay particular 
attention to possible niches that provide harborage for L. monocytogenes. 
i.  Exploratory testing 
You must conduct additional testing of surfaces and areas surrounding the area where Listeria 
spp. was detected to evaluate the extent of the problem, including the potential for Listeria spp. 
or L. monocytogenes to have become established in a niche ( 112.146(a)). Consider the 
following: 
  Exploratory testing provides you with information about the extent of the problem 
represented by the initial positive test result (e.g., whether the presence of Listeria spp. is 
isolated or more extensive). You do not need to wait for the results of the exploratory 
testing before doing any cleaning and sanitizing. However, you should use the results of 
exploratory testing to inform your decision about the extent of cleaning and sanitizing 
that you will perform as required by  112.146(b) (see next subheading); 
  If the original positive test result is from a composite sample, you should either conduct 
additional follow-up testing to identify the specific non-FCS(s) that is contaminated with 
Listeria spp. or, alternatively, conduct your exploratory testing as if each non-FCS site 
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represented by the composite is positive (i.e., conduct exploratory testing of surfaces and 
areas surrounding all of the sites represented by the composite); 
  Exploratory testing should include at least 3 samples from surrounding FCS and non-FCS 
sites in close proximity to the positive site; 
  Exploratory testing while in production: If you receive a positive Listeria spp. result for 
from a routine sample for a non-FCS site while you are in production (e.g., you are either 
still growing the production batch of sprouts that was growing when you took your 
routine sample, or you have started another production batch of sprouts), you should 
conduct exploratory testing during that production cycle;  
  Exploratory testing when not in production: If you receive a positive Listeria spp. result 
for a routine sample for a non-FCS site when you are not in production (e.g., the 
production batch of sprouts that was growing when you took your routine sample has 
been completed and you have not started the next production batch), you should conduct 
exploratory testing during the production of the next batch; and 
  If you receive any positive results from your exploratory testing, you should conduct 
intensified cleaning and sanitizing and intensified sampling and testing (discussed below 
as a recommended response to a 2nd positive) 
ii.  Cleaning and sanitizing 
You must clean and sanitize the affected surfaces, including FCSs and non-FCSs as well as 
surfaces from the surrounding area where the positive sample was taken ( 112.146(b); see also 
 112.123(d)(1)-(2); 112.143(b)). Consider the following: 
  At a minimum, the site of the initial positive Listeria spp. result and the sites of any 
additional positive results found during exploratory testing should all be cleaned and 
sanitized before your next production run; and 
  You should also consider verifying the efficacy of your cleaning and sanitizing using 
additional methods (e.g., ATP testing) beyond the required testing before your next 
production run; See Section III (Cleaning and Sanitizing); and 
  If you receive negative test results from all of your exploratory testing, and you have 
cleaned and sanitized the affected FCSs and non-FCSs from the surrounding area where 
the original positive sample was taken, you may consider the actions required by 
112.146(b) and 112.146(c) to have been accomplished while conducting the actions 
required by  112.146(a). 
b. Listeria spp. positive on non-FCS during exploratory testing or cleaning 
verification testing (2nd positive) 
i.  Intensified cleaning and sanitizing 
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If any of the exploratory testing samples are positive for Listeria spp., as an action to prevent 
recurrence of the contamination, you must clean and sanitize the affected surfaces and 
surrounding areas ( 112.146(b)). The cleaning and sanitizing that you perform under these 
circumstances should be intensified beyond your routine efforts.   Intensified cleaning and 
sanitizing include sanitation measures that you perform in addition to your normal sanitation 
procedures. See also Section III.F. (Cleaning and Sanitizing Conducted in Response to Suspected 
or Known Contamination). You should escalate the intensity of these measures in response to 
additional positive samples. Intensified cleaning and sanitizing can include: 
  Increasing the frequency of cleaning and sanitizing for certain pieces of equipment;  
  Breaking down the equipment into its parts for further cleaning;  
  The use of more effective or different cleaning and sanitizing methods or agents (e.g. 
more aggressive scrubbing, applying heat (where possible)) (see Section III (Cleaning 
and Sanitizing)); 
  Intensified (e.g., more vigorous, longer duration) scrubbing of surfaces; 
  Identifying, cleaning, and sanitizing of possible harborage sites and possible cross- 
contamination routes; and 
  Soaking of equipment parts in an appropriate sanitizing agent for an appropriate duration. 
ii.  Intensified testing 
If any of the exploratory testing samples are positive for Listeria spp., you should, as an action to 
prevent recurrence of the contamination ( 112.146(c) and 112.146(e)), conduct another round 
of sampling and testing (intensified testing), both to verify the effectiveness of your intensified 
cleaning and sanitizing, and to look for possible harborage sites in the affected area. 
To look for possible harborage sites, you should disassemble equipment as appropriate and 
sample and test areas of the equipment exposed by disassembly prior to cleaning and sanitizing 
the equipment. Consider the following: 
1)  The follow-up samples should include at least 3 samples, including the initial positive site, 
surrounding sites that tested positive in exploratory testing, and surrounding FCS and non-
FCS sites in close proximity to any positive sites; and   
2)  If your intensified sampling and testing results are all negative, you should return to routine 
environmental monitoring.  However, we recommend that you target those surfaces where 
Listeria spp. positives were previously found for sampling and testing during your next 
routine environmental sampling event. If you find another Listeria spp. positive result at the 
site of the initial positive or in any surrounding areas, you should take further steps as 
described below. 
c. Listeria spp. positive on non-FCS during intensified testing (3rd or subsequent 
positive) 
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i.  Additional activities/comprehensive investigation 
If your intensified testing results in an additional positive Listeria spp. sample(s), as an action to 
prevent recurrence of the contamination ( 112.146(e)), you should conduct additional activities 
to determine the source and route of contamination, including a comprehensive investigation, as 
discussed in Section VI. L. 3. b (Listeria spp. positive on FCS during exploratory testing (2nd 
positive). These actions may vary depending on the risk that an FCS or food could become 
contaminated from the positive non-FCS site. Examples of such actions include escalating 
mitigation efforts to identify and eliminate the Listeria spp. source and consultation with a 
Listeria control expert. 
The example in Figure 4 addresses testing and follow-up actions for a single positive finding of 
Listeria spp. on a Zone 2 non-FCS during one sampling period. Detecting Listeria spp. at several 
Zone 2 non-FCS sampling locations during the same sampling period could indicate that your 
routine sanitation procedures are inadequate and could indicate that Listeria spp. has become 
established more broadly in harborages in Zone 2. The risk associated with cross-contamination 
to an FCS (Zone 1) or food increases as the number of contaminated Zone 2 non-FCS sites 
increases. When several Zone 2 non-FCS sites test positive for Listeria spp. during one sampling 
period, we recommend that you review your written sanitation procedures to identify and 
implement more effective measures and escalate your corrective actions until the situation is 
resolved. 
   
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Figure 4. Non-FCS* Testing and Follow-Up Activities for Zone 2 Decision Tree 
 
 
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M. Analysis of Data for Trends 
To make the best use of the data that you collect through your environmental monitoring 
program, we recommend that you analyze the data (e.g., sample results, corrective actions, 
findings from comprehensive investigations) from the program over time for trends that can help 
you to continuously improve sanitation conditions and reduce the overall percentage of positive 
environmental samples in your operation. This trend analysis could reveal that L. monocytogenes 
is not being controlled in your operation (e.g., a resident strain has become established in a niche 
environment), triggering you to take additional steps to control it.   
Examples of trends that could indicate that L. monocytogenes is not being controlled are: 
  Finding Listeria spp. in the same area on multiple, but non-consecutive, sampling 
occasions (e.g., positive one week and negative the next, and then positive again a couple 
of weeks later, appearing to be isolated positives); 
  An increase in the overall percentage of Listeria spp. positive samples in your operation; 
and 
  Increases in the percentage of Listeria spp. positive environmental samples in particular 
areas of your operation. 
Even if you have taken appropriate corrective actions for individual positive test results from a 
particular site, the continued finding of Listeria spp. positives at that site or in that area over time 
may indicate a continuing problem, such as an unidentified harborage site. If your analysis of 
data indicates a potential problem, such as an increased incidence of Listeria spp. in your 
operation, you should conduct a more complete investigation to determine if further actions are 
warranted and take appropriate corrective actions to reduce the incidence of Listeria spp. in your 
operation. A trend analysis may also lead you to update your written environmental monitoring 
plan.  
VIII. Recordkeeping 
Recordkeeping is an essential component of safely growing, harvesting, packing, and holding 
food, including sprouts. Records can be used to ascertain safety of products, monitor storage 
conditions, document good agricultural practices, and track receipt and distribution of product(s). 
Records also offer written evidence that required conditions are being consistently met and your 
established procedures are being properly followed and are under control (e.g., by facilitating 
trend analyses). They may also be used to help determine the cause of underlying problems in 
the event of a foodborne illness outbreak or recall. 
In this section, we provide a brief overview of the recordkeeping requirements in the Produce 
Safety Rule for sprout operations covered by Subpart M.  
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A. Recordkeeping Overview 
1. General requirements 
Except as otherwise specified, the requirements of Subpart O apply to all records that are 
required under the Produce Safety Rule. You must include the following in all required records, 
as applicable and unless otherwise specified: 
  The name and location of your sprout operation ( 112.161(a)(1)(i)); 
  The actual values and observations obtained during monitoring ( 112.161(a)(1)(ii)); 
  An adequate description (such as the commodity name, or the specific variety or brand 
name of a commodity, and, when available, any lot number or other identifier) of covered 
produce applicable to the record ( 112.161(a)(1)(iii)); 
  The location of a growing area or other area (for example, a specific growing bin) 
applicable to the record ( 112.161(a)(1)(iv)); and 
  The date and time of the activity documented (e.g., date and time of sample collection) 
(112.161(a)(1)(v)).  
Required records must also: 
  Be created at the time an activity is performed or observed ( 112.161(a)(2)); 
  Be accurate, legible, and indelible ( 112.161(a)(3));  
  Be dated, and signed or initialed by the person who performed the activity documented ( 
112.161(a)(4)); and 
  Be reviewed, dated, and signed, within a reasonable time after the records required under 
112.7(b), 112.30(b)(2), 112.50(b)(2), (4), and (6), 112.60(b)(2), 112.140(b)(1) and (2), 
and 112.150(b)(1), (4), and (6) are made, by a supervisor or responsible party 
(112.161(b)). 
Records can take many forms, including photographs. However, the photos alone do not have 
sufficient information to meet the general requirements for a record, and the additional 
information listed above must be captured (see  112.161).   
a. Actual values and observations 
Section 112.161(a)(1)(ii) requires records to include, as applicable, the actual values and 
observations obtained during monitoring. Actual values and observations mean that: (a) the 
value or observation, as applicable, itself is written (e.g., 96F) rather than a summary term, such 
as OK or Passed; and (b) truthful information is recorded. For example, if you are 
documenting a repair as a corrective action, we recommend taking pictures before and after the 
repair. Another example of the recording of actual values and observations is the listing of food 
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and/or food contact surfaces (FCSs) that may have been affected by Listeria contamination, as 
identified during a comprehensive investigation as a corrective action following repeated Listeria 
spp. positive results on an FCS.    
b. Date and time 
As applicable, you must include the date and time of the documented activity on the records ( 
112.161(a)(1)(v)). Records must also be created at the time that an activity is performed or 
observed ( 112.161(a)(2)). You must not pre-fill records, nor rely on your memory to write 
down the information later. For example, pertinent information, such as the date and time your 
seed treatment was applied, spent sprout irrigation water sample was collected, and spent sprout 
irrigation water sample was received and analyzed by the testing lab must be recorded at the time 
that the action is taken. 
c. Adequate description of covered produce applicable to the record 
You must include an adequate description of covered produce to which the record applies ( 
112.161(a)(1)(iii)). For sprouts, we recommend including a production batch lot number that 
includes the product name (e.g., sandwich blend), variety (e.g., alfalfa sprouts and/or clover 
sprouts), and brand name that your operation uses to identify the product. We recommend that 
you assign a unique identifier for each individual production batch of sprouts (as that term is 
defined in  112.3). Using unique production batch numbers and including them on all applicable 
records enables you to track a production batch of sprouts internally and through distribution in 
the event there is a problem with that production batch.  
If you use an identification system for product after it has been packaged for sale that is different 
from your production batch code, your ability to track the product will be enhanced if you are 
able to link the packed product code to the production batch of sprouts used to produce that 
packed product. For example, you may combine product from Production Batch A with 
product from Production Batch B during packing and assign product identifier #1234 to the 
final packed product. In this example, your ability to track the product would be enhanced if you 
are able to link packed product identifier #1234 to Production Batches A and B.9 
We recommend that your records include the following information, as applicable: 
  The seed lot number(s) for the seeds used to grow the production batch of sprouts (so that 
the lot of seeds that were used to grow each production batch of sprouts can be easily and 
reliably identified in the event of a problem with the production batch of sprouts); 
  Container size/type; 
 
9 FDA published a rule to establish additional traceability recordkeeping requirements (beyond what is already 
required in existing regulations) for persons who manufacture, process, pack, or hold foods the Agency has 
designated for inclusion on the Food Traceability List. The rule will affect some sprout operations. For further 
information, see Food Traceability List at https://www.fda.gov/food/food-safety-modernization-act-fsma/food-
traceability-list. 
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  Date packaged; 
  Number of units packaged; 
  Holding area; and 
  Any other comments or information that may be useful in the event of a problem with the 
production batch of sprouts. 
We also recommend that, for records of seed treatment that you perform at your operation to 
reduce microorganisms of public health significance (as required by  112.142(e)(1) and 
112.150(b)(1)), you include a description of the seeds to which the record relates. Such a 
description should include the type of seeds treated, and the seed lot number. 
d. Location of growing area or other area applicable to the record 
You must include the location of the growing area or other area applicable to the record ( 
112.161(a)(1)(iv)). For example, you could include: in your records for testing spent sprout 
irrigation water, the specific sprouting room and/or growing unit in which you grew the batch of 
sprouts being tested (e.g., Bean Sprouting Room #2, Cabin 1); in your records for seed treatment, 
the specific seed treatment area in which you treated the seeds; or, in your records of 
environmental monitoring, the particular packing line from which the sample was collected. If 
you have a diagram of your sprout operation, you may find it helpful for the names of the 
locations in the records to match the names in your diagrams. 
e. Accuracy, legibility, indelibility 
Required records must be accurate, legible, and indelible ( 112.161(a)(3)). For example, if you 
make a mistake on a record, you should mark through the mistake with a single line, add the 
correct information as close as possible to the information entered in error and initial and date the 
correction. You should not write over the existing information, obscure it by scratching it out, or 
use liquid correction fluid. While handwriting styles may vary, the information on a required 
record must be legible so that company officials and regulatory agencies, as appropriate, can 
review it. You must document required information in an indelible manner (e.g., in ink) so that it 
cannot be erased. 
 f. Dated, and signed or initialed by the person who performed the activity 
The individual who performed the activity documented must date, and sign or initial the required 
record ( 112.161(a)(4)). Examples of such individuals include the person inspecting the 
agricultural water system (for records required under  112.50(b)(1)), the worker who sanitized 
equipment used for growing sprouts (for records required under  112.140(b)(1)), or the 
laboratory analyst who tested spent sprout irrigation water samples for microbiological 
contamination (for records required under  112.150(b)(4)). 
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2. Duplication not required 
Section 112.163(a) provides that you are not required to duplicate any existing records if those 
records contain all of the required information and satisfy the requirements of the Produce Safety 
Rule. Similarly, if you have records containing some but not all of the required information,  
112.163(b) provides you the flexibility to keep any new information required either separately or 
combined with your existing records, even where the formats for the records may not be the 
same. For example, if you already receive an annual report of your municipal water quality from 
your county/city, this record may meet the requirement at  112.46(a)(2) for such public water 
system results, and you would not need obtain further certification.   
3. Record retention and availability 
You must keep required records for at least 2 years past the date the record was created ( 
112.164(a)(1)). 
You must keep records that relate to the general adequacy of equipment or processes or records 
that relate to analyses, sampling or action plans that you use, including the results of scientific 
studies, tests, and evaluations, for at least 2 years after you discontinue use of  such equipment or 
processes, or records related to analyses, sampling, or action plans ( 112.164(b)). Examples of 
such records for a sprout operation include: the written sampling plan for spent sprout irrigation 
water or sprouts, including the corrective action plan ( 112.147(a) and (c) and 112.150(b)(3)); 
the written environmental monitoring plan including the corrective action plan ( 112.145(a) 
and (e) and 112.150(b)(2)); and documentation of scientific data or information relied on to 
support the adequacy of a method used to satisfy the requirements of  112.43(a)(1) and (a)(2) 
for treating agricultural water ( 112.50(b)(3)).  
If you are eligible for the qualified exemption in accordance with  112.5, you must retain 
records that you rely on during the 3-year period preceding the applicable calendar year to satisfy 
the criteria for a qualified exemption, in accordance with  112.5 and 112.7, as long as 
necessary to support your exemption status during the applicable calendar year ( 112.164(a)(2)). 
In addition, you must have all required records readily available and accessible during the 
retention period for inspection and copying by FDA upon oral or written request, except that you 
have 24 hours to obtain records you keep offsite and make them available and accessible for 
inspection and copying ( 112.166(a)). Offsite storage of required records is permissible, 
provided such records  can be retrieved and provided onsite within 24 hours of request for 
official review ( 112.162(a)). Electronic records are considered to be onsite at your sprout 
operation if they are accessible from an onsite location at your farm ( 112.162(b)). 
4. Format 
As required by  112.165, you must keep records as: (1) original records; (2) true copies; or (3) 
electronic records. Original records generally refers to the document where the information 
was first recorded. True copies include, for example, photocopies, pictures, scanned copies, 
microfilm, microfiche or other accurate reproductions of the original records. True copies of 
records should be of sufficient quality to detect whether the original record was changed in a 
manner that obscured an original entry (e.g., through the use of liquid correction fluid). 
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Electronic records are subject to the same requirements as paper records. We do not require 
electronic records, nor are we specifying the form or format of the records that must be 
established and maintained, except as otherwise set forth in Subpart O (e.g., certain content is 
required when applicable as discussed in Section VII. A. 1 (General Requirements) above). To 
satisfy the requirements of the Produce Safety Rule, paper or electronic records, or a 
combination of the two, may be used. 
Records that are established or maintained to satisfy the requirements of part 112 and that meet 
the definition of electronic records in  11.3(b)(6) are exempt from the requirements of part 11 
(Electronic Records; Electronic Signatures). Records that satisfy the requirements of part 112, 
but that also are required under other applicable statutory provisions or regulations, remain 
subject to part 11 ( 112.165(c)). 
Although part 11 does not apply, except for records otherwise subject to part 11 (as provided in  
112.165(c)), you should take appropriate measures to ensure that electronic records are 
trustworthy, reliable, and generally equivalent to paper records with handwritten signatures 
executed on paper.   
B. Supervisory Review of Records 
Certain required records must be reviewed, dated, and signed by a supervisor or a responsible 
party within a reasonable time after the records are created ( 112.161(b)). These records 
include, as applicable: 
  Records related to eligibility for the qualified exemption ( 112.7(b)); 
  Records related to required training of personnel ( 112.30(b)); 
  Documentation of the results of all agricultural water testing ( 112.50(b)(2)); 
  Documentation of the results of water treatment monitoring ( 112.50(b)(4)); 
  Documentation of actions you take when your agricultural water does not meet the water 
quality requirements (112.50(b)(6)); 
  Documentation that process controls were achieved for a treated biological soil 
amendment of animal origin you produce for your own covered sprout operation(s) ( 
112.60(b)(2)); 
  Documentation of cleaning and sanitizing of equipment ( 112.140(b)(1) and (2)); 
  Records related to seed treatment ( 112.150(b)(1)); 
  Documentation of analytical test results for all sprout-specific testing (including spent 
irrigation water testing and environmental monitoring) done under Subpart M ( 
112.150(b)(4)); and, 
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  Documentation of sprout-specific corrective actions you take under Subpart M ( 
112.150(b)(6)). 
The person reviewing records should first make sure that all the required records are kept and are 
complete, ensure that they were completed accurately and in a timely manner, consider whether 
the records suggest any problems that need to be corrected, and institute any corresponding 
corrective actions, as necessary. The reviewer should also ensure that the observations and 
results are consistent with the expected range for the activity, look for any trend that could lead 
to problems in the future if not adequately addressed, and institute preventive measures as 
necessary. For example, a supervisor reviews a seed treatment record for seed to which 19,000 
ppm sodium hypochlorite should be applied and available chlorine concentration monitored to 
ensure proper treatment. However, the supervisor notes that the recorded value of sodium 
hypochlorite is 500 ppm. The supervisor should follow up with the worker who made the 
recording to determine whether there was a malfunction with the monitoring instrument, a 
difficulty by the worker in reading the instrument, a recording error (i.e., the record is not 
accurate), or an issue of not following the seed treatment protocol. 
C. Sprouts-Specific Record Requirements and Recommendations 
1. Enforcement Discretion for Requirements of Subpart M (applies to soil or substrate 
grown sprout types that are harvested by the customer before use) 
If you produce sprouts that will be sold to a commercial entity in a tray that was used for 
growing the sprouts, that contains the soil/substrate and roots intact, and such sprouts are then to 
be cut above the soil and/or substrate line at the retail or other establishment immediately before 
use, we intend to exercise enforcement discretion for the requirements of Subpart M provided 
you annually collect written assurances from your customers, stating that the sprouts will be cut 
above the soil and/or substrate line before use. 
2. Records for Seeds for Sprouting 
a. Required records for seed for sprouting 
Section 112.150(b)(1) requires that you establish and keep documentation of your treatment of 
seeds to reduce microorganisms of public health significance in the seeds, at your sprout 
operation; or alternatively, documentation (such as a Certificate of Conformance) from your seed 
supplier that seeds are treated to reduce microorganisms of public health significance and are 
appropriately handled and packaged following the treatment, in accordance with the 
requirements of  112.142(e). Like all records  required by the Produce Safety Rule, seed 
treatment records must comply with all applicable  requirements for records in Subpart O. For 
example, as applicable, these records must include:  
  The name and location of your sprout operation ( 112.161(a)(1)(i)); 
  Actual values and observations obtained during monitoring ( 112.161(a)(1)(ii)) (e.g., 
observations of treatment conditions and key parameters monitored during preparation 
and application of treatments (e.g., chemical composition of treatment solution, heating 
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temperature, equipment used, concentration of treatment solution, treatment time, pH of 
treatment solution, presence of agitation));  
  An adequate description of the covered produce ( 112.161(a)(1)(iii)) (e.g., the type of 
seed being treated, and, when available, any lot number or other identifier of the seed 
applicable to the record);   
  The location of the area applicable to that record ( 112.161(a)(1)(iv)) (e.g., the seed 
treatment room);  
  The date and time of the activity documented ( 112.161(a)(1)(v)) (e.g., the date and time 
the treatment was applied); and 
  This record must be dated and signed or initialed by the person who performed the 
activity documented ( 112.161(a)(4)) and must be reviewed, dated and signed within a 
reasonable time after the records are made by a supervisor or responsible party ( 
112.161(b)).  
If you are relying on prior treatment of seeds by someone in the supply chain prior to your 
operation (e.g., a grower, distributor, or supplier of seeds), whether to fulfill the treatment 
requirement completely or for the purpose of considering such prior treatment when applying 
appropriate additional treatment at your operation in accordance with  112.142(e)(2), you must 
obtain documentation (such as a Certificate of Conformance) from the grower, distributor, or 
supplier of seeds that the treatment was conducted using a scientifically valid method to reduce 
microorganisms of public health significance and that the treated seeds were handled and 
packaged, following the treatment, in a manner that minimizes the potential for contamination ( 
112.150(b)(1). Such records must comply with all applicable  requirements for records in Subpart 
O.  For example, as applicable, these records must: 
  Be dated, and signed or initialed by the person who performed the activity documented ( 
112.161(a)(4)), e.g., a person from the grower, distributor or supplier of seeds that 
applied the seed treatment; and  
  Be reviewed, dated and signed within a reasonable time after the records are made by a 
supervisor or responsible party ( 112.161(b)). For example, a supervisor or responsible 
party at the sprout operation must review, date and sign the Certificate of Conformance 
received for prior treatment of seeds by the grower, distributor or supplier of seeds.  
Section 112.150(b)(6) requires that you establish and keep records of certain corrective actions. 
These include those corrective actions taken in accordance with  112.142(b) and (c) in 
response to contaminated seed and  112.148. Such records must comply with all applicable 
requirements for records in Subpart O. For example, as applicable, these records must include: 
  The name and location of your sprout operation ( 112.161(a)(1)(i)); 
  Actual values and observations obtained during monitoring ( 112.161(a)(1)(ii)) (e.g., 
date/time, and person who provided the notification to the seed supplier, the information 
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provided, and responses from the seed supplier after notification); if conducting follow-
up actions to investigate the potential source of contamination, the actual findings of 
those follow-up actions; if you treat an affected lot of seeds with a process that is 
reasonably certain to achieve destruction or elimination in seeds of the most resistant 
microorganisms of public health significance, the records you keep to document this 
action must include the same content required for records documenting routine seed 
treatment (see above in this same section). Further, our recommended content for records 
related to routine seed treatment is also applicable to these records; 
  An adequate description of the covered produce ( 112.161(a)(1)(iii)) (e.g., the type of 
seed affected, and, when available, any lot number or other identifier of the seed to which 
the record is applicable);  
  The location of the area applicable to that record ( 112.161(a)(1)(iv)) (e.g., the seed 
treatment room if treating the seeds in compliance with  112.142 (e)(1), or the areas of 
the sprout operation that were included in an investigation as part of a follow-up action 
undertaken in compliance with  112.142(c)(2)); and  
  The date and time at which you performed each activity ( 112.161(a)(1)(v)) (e.g., 
notification of seed supplier, returning the affected seed lot to your supplier, destroying 
sprouts grown from the affected seed lot, treating seeds with a process reasonably certain 
to achieve destruction or elimination in seeds of the most resistant microorganisms of 
public health significance, or conducting follow up actions to investigate the potential 
source of the contamination). 
Section 112.150(b)(4) requires that you establish and keep records of all analytical tests 
conducted for purposes of compliance with Subpart M. This includes records of test results from 
any testing conducted as part of follow-up actions related to suspected contamination of a seed 
lot with a pathogen in compliance with  112.142(c)(2) (see also  112.150(b)(6)).  
b. Recommended records for seeds for sprouting 
While not requirements of the Produce Safety Rule, we recommend these practices as prudent 
for seeds used for sprouting.  
  To ensure seed treatments (as required under  112.142(e)) are consistently applied 
correctly, if you treat seeds at your sprout operation, you should develop a written seed 
treatment Standard Operating Procedure (SOP). See Section V (Seeds for Sprouting) for 
more information.  
  We recommend you develop a seed receiving program setting out the SOPs you follow to 
receive and inspect seed upon receipt and the criteria you use in determining whether to 
accept a shipment. We recommend that you use a seed receiving and inspection checklist 
for all incoming seed shipments to record and maintain information about the lot. An 
example of a seed receiving and inspection checklist can be found in Section V. A. 3 
(Visual Inspection of Seeds and Their Packaging). 
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  To minimize the risk of contamination associated with seeds for sprouting, we 
recommend that you request copies of assurances from your seed suppliers that the seeds 
they provide were produced under GAPs and handled according to food safety best 
practices throughout harvesting, conditioning, storage, and transportation (Ref. 17). We 
recommend receiving and maintaining copies of assurances from your seed suppliers as a 
prudent practice when sourcing the seeds used for sprouting and as a practice to support  
112.142(a). 
  We recommend that you establish and keep sufficient records to allow you to maintain 
the lot identity of the seeds you receive and the sprouts you produce. We recommend that 
you maintain such records related to each lot of seeds for at least two years after your last 
use of the seed lot. See subsection C (Corrective Actions for Seeds That May be 
Contaminated with a Pathogen) of Section V (Seeds for Sprouting) for more information. 
Seed lot numbers are typically assigned by seed suppliers. The seed lot number may appear on 
seed packages, or on seed shipment records. The seed lot number allows both the seed supplier 
and you to track seeds. Although not required by the Produce Safety Rule we recommend that 
you keep sufficient records to connect your test results and corrective actions to specific seed lots 
whenever possible. In addition, FDA published a rule to establish additional traceability 
recordkeeping requirements (beyond what is already required in existing regulations) for persons 
who manufacture, process, pack, or hold foods the Agency has designated for inclusion on the 
Food Traceability List. The rule will affect some sprout operations. For further information, see 
Food Traceability List at https://www.fda.gov/food/food-safety-modernization-act-fsma/food-
traceability-list.   
3. Required Records for Spent Sprout Irrigation Water  
Sections 112.150(b)(3) and 112.147(a) require that you establish and implement a written 
sampling plan that identifies the number and location of samples (of spent sprout irrigation water 
or sprouts) to be collected for each production batch of sprouts to ensure that the collected 
samples are representative of the production batch when testing for contamination. 
  The plan should identify: the sample type (e.g., spent sprout irrigation water, sprouts); 
pathogens to be tested; specific test methods to be used; person(s) responsible for 
collecting the sample; timing of sample collection (e.g., number of hours after start of 
growing cycle); procedures for aseptic technique; sampling tools to be used; laboratory to 
be used; and records to be kept). 
  Your sampling plan must include a corrective action plan, which must, at a minimum, 
require you to take the actions specified in  112.148, and must detail when and how you 
will accomplish those actions, if the samples of spent sprout irrigation water or sprouts 
test positive for E. coli O157:H7, Salmonella spp., or a pathogen meeting the criteria in  
112.144(c). (see Section V (Sampling and Testing of Spent Sprout Irrigation Water (or 
In-Process Sprouts)) of the draft re-issued sections (Ref. 18) for additional information on 
the written sampling plan) 
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Section 112.150(b)(5) requires you to establish and keep documentation of any analytical 
methods used to test for E. coli O157:H7 or Salmonella species in lieu of the methods for sprout 
production batch testing that are incorporated by reference in  112.152 and 112.153.  
  Such records should include: 1) the name and identification number of the method 
validated by a third party methods validation organization, or 2) the detailed analytical 
procedures and information relevant to the determination of the scientific validity of the 
alternate method;  
  In addition, if you test spent sprout irrigation water (or in-process sprouts) for any other 
pathogen(s) meeting the criteria in  112.144(c), you should keep records of the 
scientifically valid test method you used for such testing ( 112.153(b)). 
Section 112.150(b)(4) requires that you establish and keep records of all analytical tests 
conducted of spent sprout irrigation water or in-process sprouts for Salmonella spp., E. coli 
O157:H7 as required under  112.144(b), and any additional pathogen required under  
112.144(c). The results must be documented whether they were conducted by a third party or 
your own (e.g., in-house) laboratory. Records of required pathogen test results for spent sprout 
irrigation water (or sprouts) must comply with all applicable  requirements for records in Subpart 
O. For example, as applicable, these records must include: 
  The name and location of your sprout operation ( 112.161(a)(1)(i)); 
  Actual values and observations obtained (e.g., test results from spent sprout irrigation 
water (or sprouts) for each pathogen, including screening and confirmatory test results (if 
conducted)) ( 112.161(a)(1)(ii)); 
  An adequate description of covered produce applicable to the record (e.g., production 
batch number) ( 112.161(a)(1)(iii)); 
  Location of the growing area from which the sample was collected (e.g., growing unit 
and growing area/room information) ( 112.161(a)(1)(iv));  
  Date and time of activity documented (e.g., date and time of sample collection, and 
sample receipt and analysis by the testing lab) ( 112.161(a)(1)(v)); and 
  In addition, such records must be dated, and signed or initialed by the person who 
performed the activity documented (e.g., the individual who conducted sample analysis ( 
112.161(a)(4)), and reviewed, dated, and signed, within a reasonable time after the 
records are made, by a supervisor or responsible party ( 112.161(b)). 
Section 112.150(b)(6) requires that you establish and keep records of corrective actions you take 
in accordance with  112.142(b) and (c), 112.146 and 112.148. See subsection V. G. 3 
(Documenting your corrective actions) in Section V (Sampling and Testing of Spent Sprout 
Irrigation Water (or In-Process Sprouts)) in the draft re-issued sections (Ref. 18).  
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Such records also must comply with all applicable  requirements for records in Subpart O.  For 
example, as applicable, these records must include: 
  The name and location of your sprout operation ( 112.161(a)(1)(i)); 
  Actual values and observations (e.g., see above Required Records for Seed for Sprouting 
for examples related to the seed lot; sanitizer concentration, post-cleaning and sanitizing 
ATP test results, observations related to actions taken to prevent reoccurrence of 
contamination (e.g., employee retraining (see  112.21(d)) and observation for 
compliance with procedures)) ( 112.161(a)(1)(ii));  
  An adequate description of covered produce applicable to the record (e.g., affected sprout 
production batch number) ( 112.161(a)(1)(iii)); 
  Location of a specific growing area or other area applicable to the record (e.g., location or 
other identifiers for growing units and other surfaces cleaned and sanitized as part of 
corrective actions) ( 112.161(a)(1)(iv)); 
  Date and time of activity documented (e.g., when cleaning and sanitizing or employee re-
training corrective actions were conducted) ( 112.161(a)(1)(v)); and 
  In addition, such records must be dated, and signed or initialed by the person who 
performed the activity documented (e.g., the individual who reported positive test 
findings to the seed supplier; the individual who took steps to ensure a contaminated 
production batch of sprouts did not enter commerce ( 112.161(a)(4)) and reviewed, 
dated, and signed within a reasonable time after the records are made, by a supervisor or 
responsible party ( 112.161(b)).   
4. Required records for environmental monitoring 
Sections 112.150(b)(2) requires that you establish and keep a written environmental monitoring 
plan, in accordance with section 112.145, which is designed to identify Listeria monocytogenes 
if it is present in the growing, harvesting, packing, or holding environment.  
  Your environmental monitoring plan must be directed to sampling and testing for either 
Listeria species or L. monocytogenes and your written environmental monitoring plan 
must include a sampling plan, including what you will test collected samples for, how 
often you will collect environmental samples, and a list of sample collection sites  
112.145(b), 112.145(c)). Your written environmental monitoring plan must include a 
corrective action plan that, at a minimum, requires you to take the actions in  112.146, 
and details when and how you will accomplish those actions, if the growing, harvesting, 
packing, or holding environment tests positive for Listeria species or L. monocytogenes. 
  The plan should identify the specific test methods to be used, the person(s) responsible 
for collecting the sample, sample collection method type (e.g., sponge vs. swab sampling, 
whether any samples will be composited), procedures for aseptic technique, laboratory to 
be used, and records to be kept. 
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  Section 112.150(b)(5) requires that you establish and keep records of any analytical 
methods you use in lieu of the methods that are incorporated by reference in 112.152(a). 
For example, your records could specify that you are using one of the methods that FDA 
listed in Equivalent Testing Methodologies for Listeria species and Listeria 
monocytogenes in Environmental Samples on our Internet site 
(https://www.fda.gov/food/laboratory-methods-food/equivalent-testing-methodologies-
listeria-species-and-listeria-monocytogenes-environmental-samples). If the alternate 
method that you use is not listed by FDA, your records of any alternate analytical 
methods you use in lieu of the methods that are incorporated by reference in  112.152(a) 
( 112.150(b)(5)) should include 1) name and identification number of the method 
validated by a third party methods validation organization, or 2) the detailed analytical 
procedures, an explanation of why the alternate method is at least equivalent to the 
reference method (such as a scientific study, or data or information regarding the 
sensitivity, specificity, and precision of the alternate method compared to the reference 
method), and any other relevant information supporting the use of the alternate method. 
(see Section VI (Environmental Monitoring) of this guidance for additional information 
on the written environmental monitoring plan). 
Section 112.150(b)(4) requires that you establish and keep records of test results from 
environmental testing for Listeria spp. or L. monocytogenes you perform in compliance with  
112.144(a) and 112.145 or any additional testing conducted if you detect Listeria spp. or L. 
monocytogenes in the environment in compliance with  112.146 (a), (c), (d), and (e). The results 
of all analytical tests must be documented, regardless of whether they were conducted by your 
own (e.g., in-house) laboratories or third-party laboratories. Records of environmental 
monitoring tests must comply with all applicable  requirements for records in Subpart O. For 
example, as applicable, these records must include: 
  The name and location of your sprout operation ( 112.161(a)(1)(i)); 
  Actual values and observations obtained (e.g., test results, including screening and 
confirmatory test results (if conducted), and any results for Listeria spp. and L. 
monocytogenes) ( 112.161(a)(1)(ii)); 
  An adequate description of covered produce applicable to the record (e.g., production 
batch number(s) of sprouts in production at the time of the environmental sample) ( 
112.161(a)(1)(iii)); 
  Location of the growing area or other area(s) applicable to the record (e.g., identification 
of each sampling site, including whether it is an FCS or non-FCS site, description of the 
room or area in the sprout operation) ( 112.161(a)(1)(iv)); and 
  Date and time of the activity documented (e.g., date/time of sample collection, and 
sample receipt and analysis by the testing lab) ( 112.161(a)(1)(v)). 
Section 112.150(b)(6) requires that you establish and keep records of corrective actions 
conducted in accordance with the requirements of  112.146. See Section VI (Environmental 
Monitoring) for more information. Such records must include: 
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  The name and location of your sprout operation (112.161(a)(1)(i)); 
  Actual values and observations obtained (e.g., observations and information related to a 
comprehensive investigation following repeated Listeria spp. positive results on an FCS, 
such as the food and FCSs potentially affected by the contamination event) ( 
112.161(a)(1)(ii)); 
  An adequate description of covered produce applicable to the record (e.g., production 
batch number of sprouts in production at the time of the corrective action) ( 
112.161(a)(1)(iii)); 
  Location of a growing area or other area(s) applicable to the record (e.g., identification of 
locations sampled or subjected to intensified cleaning and sanitizing) ( 
112.161(a)(1)(iv)); and 
  Date and time of the activity documented (e.g., date/time of testing or intensified cleaning 
and sanitizing) ( 112.161(a)(1)(v)).  
6.  Records Required by Provisions Other Than Subpart M 
The records required under the Produce Safety Rule are dependent, in part, on the nature of the 
practices and procedures related to the covered activities in your sprout operation and are listed 
here under the applicable section of the Produce Safety Rule. Other required records that are 
relevant to sprout operations are discussed below. 
Table 6. Records Required by Provisions Other Than Subpart M 
Record Type  Description 
Records relating to  Section 112.2(b) requires sprout operations relying on the exemption for 
commercial  produce that receives commercial processing that adequately reduces the 
processing  presence of microorganisms of public health significance, provided for 
exemption  in  112.2(b), to establish and maintain documentation of: 1) their 
required disclosures to customers; and 2) annual written assurances 
obtained from customers. 
Records relating to  Section 112.7 requires sprout operations eligible for the qualified 
eligibility for  exemption, provided for in  112.5, to establish and keep adequate 
qualified  records necessary to demonstrate that the sprout operation satisfies the 
exemption  criteria for a qualified exemption (e.g., dated sales receipts), including a 
written record reflecting that the owner, operator, or agent in charge of 
the sprout operation has performed an annual review and verification of 
the sprout operations continued eligibility for the qualified exemption. 
Records relating to  Section 112.30 requires you to establish and keep records that document 
training  required training of personnel, required by  112.21 and 112.22, 
including the date of training, topics covered, and the persons(s) trained. 
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Record Type  Description 
Records related to  Section 112.50(b)(1) requires you to establish and keep records of your 
Agricultural water  Agricultural  water system inspection findings, required by  112.42(a); 
Section 112.50(b)(2) requires you to establish and keep documentation 
of the results of all analytical tests conducted on agricultural water for 
purposes of compliance with Subpart E; 
Section 112.50(b)(3) requires you to establish and keep documentation 
of scientific data or information you rely on to support the adequacy of 
a method used to satisfy the requirements of  112.43(a)(1) and (a)(2) 
for treating agricultural water.  
Section 112.50(b)(4) requires you to establish and keep documentation 
of results of water treatment monitoring under  112.43(b).  
Section 112.50(b)(6) requires you to establish and keep documentation 
of actions you take in accordance with 112.45 (measures taken if 
agricultural water does not meet the requirements of  112.41 or  
112.44). 
Section 112.50(b)(7) requires you to establish and keep annual 
documentation of the results or certificates of compliance from a Public 
Water System required under  112.46(a)(1) or (a)(2), if  applicable.; 
and 
Section 112.50(b)(9) requires you to establish and keep documentation 
of any analytical methods that you use for water testing in lieu of the 
method that is incorporated by reference in  112.151(a). 
Records related to  Section 112.60 requires you to establish and keep certain documentation 
biological soil  relating to any treated biological soil amendments of animal origin 
amendments of  (BSAAO) you use (e.g., substrates). The required records differ based 
animal origin  on whether treatment was conducted by you or by a third party. For a 
treated BSAAO supplied by a third party, you are required to establish 
and keep documentation (such as a Certificate of Conformance) at least 
annually that the process used to treat the BSAAO is a scientifically 
valid process that has been carried out with appropriate process 
monitoring, and that the BSAAO has been handled, conveyed, and 
stored in a manner and location to minimize the risk of contamination by 
an untreated or in process BSAAO ( 112.60(b)(1)). For a treated 
BSAAO you produce for your own covered sprout operation(s), you are 
required to establish and keep documentation that process controls were 
achieved ( 112.60(b)(2)). 
Records of  Section 112.140 requires you to establish and keep documentation of the 
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Record Type  Description 
cleaning and  date and method of cleaning and sanitizing of equipment subject to 
sanitizing  Subpart L used in growing operations for sprouts and harvesting, 
packing, or holding for sprouts and other covered produce. 
VIII. Appendices 
Appendix 1. Aseptic Sampling 
You must aseptically collect environmental samples ( 112.145(d)) and samples of spent sprout 
irrigation water or sprouts ( 112.147(b)). If you are required to test your agricultural water ( 
112.44(a), 112.46(c)), you must aseptically collect agricultural water samples as well ( 
112.47(b)). Aseptic sampling is a sampling technique used to assure that the microbial load of a 
sample is not affected by the sampling method and/or the sample collector does not contaminate 
the source from which the sample is collected (including cross-contamination between sample 
sites). The use of sterile sampling implements and containers, and a prescribed sampling method, 
defines aseptic sampling (See 80 FR 74450 and references cited therein). Collected samples 
should also be handled in a manner to ensure samples are not contaminated during storage or 
during transportation to the laboratory. 
Sterile Equipment 
 
The requirements in  112.47(b),112.145(d) and 112.147(b) to collect samples aseptically 
mean that you must use sterile sampling equipment to collect the required samples. Note that 
sterile is not equivalent to clean and sanitized. Sterilization achieves a higher standard than 
cleaning and sanitizing. Sterilization refers to a validated process used to render a product free 
of all forms of viable microorganisms. In many cases, thermal methods, such as steam, are used 
to achieve sterilization (See Liquid Chemical Sterilization) (Ref. 96). Sterile refers to the end 
point achieved by a sterilizing process. You may purchase pre-packaged sterilized 
tools/equipment to use in sampling, or you may use (and re-use) tools and equipment for 
sampling that have been properly sterilized, such as in an autoclave or a dry heat oven. An 
autoclave is a piece of equipment that sterilizes instruments and equipment by using highly 
pressurized saturated steam to achieve temperatures that effectively kill microorganisms. If you 
decide to use an autoclave, any responsible personnel should receive adequate training prior to 
the use of the autoclave. When used properly, autoclaves are safe and highly effective to use to 
sterilize sample containers or sampling equipment (e.g., cups or tongs) and can be cost-effective. 
Sampling equipment, such as stainless steel forceps, spatulas, and sample containers, can be 
sterilized using an autoclave (steam heat), for example, at 121 C (250 F) for 30 minutes at 15 
psi, or for heat-resistant, dry materials in a dry-heat oven, for example, at 140 C (284F) for 3 
hours (Ref. 97). If you choose to use an autoclave, you should follow the instructions provided 
by the manufacturer to ensure that sterilizing of sampling tools and equipment is effective. If you 
choose to sterilize your own sampling tools and equipment, you should package them prior to 
sterilization in a manner to prevent contamination post-sterilization (e.g., wrap them with 
aluminum foil or Kraft paper, or placed in other suitable containers), and should only open their 
packaging immediately prior to use. 
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We recommend that you include your plans regarding use of sterilized sampling equipment in 
your written sampling plan for spent sprout irrigation water (or sprouts) (see Section V 
(Sampling and Testing of Spent Sprout Irrigation Water (or In-Process Sprouts)) in the draft re-
issued sections (Ref. 18)) and your written environmental monitoring plan (see Section VI 
(Environmental Monitoring)). 
General Aseptic Sample Collection Procedures 
 
The requirements in  112.47(b), 112.145(d), and 112.147(b) to collect samples aseptically 
also mean that, in addition to using sterile equipment for sampling, you must use a sampling 
method that does not affect the microbial load of the sample collected and does not contaminate 
the source from which the sample is collected. We recommend that you include your procedures 
for aseptic technique in your written sampling plan for spent sprout irrigation water (or sprouts) 
(see Section V (Sampling and Testing of Spent Sprout Irrigation Water (or In-Process Sprouts)) 
in the draft re-issued sections (Ref. 18)) and your written environmental monitoring plan (see 
Section VI (Environmental Monitoring)). We recommend the following aseptic techniques, 
which are generally applicable to any kind of sampling: 
1.  For activities in fully enclosed buildings, a sample collector should wear a clean lab coat, 
sterile gloves, and a hair net to ensure he or she does not contaminate the samples. 
2.  Hands should be washed immediately before sampling, and prior to putting on sterile  
gloves. Gloves should be put on in a manner that does not contaminate the outside of the 
glove. Gloves should be properly disposed of after use. 
3.  To prevent cross contamination, gloves should be changed between samples. In addition, 
you should change gloves if you touch any surfaces other than the sample sites, such as 
garbage, drains, or the floor. 
4.  Hands should be kept away from mouth, nose, eyes, and face while collecting samples. 
You should not cough or sneeze in the vicinity of the samples, and if you do, discard the 
affected sample(s). 
5.  Sampling instruments should be protected from contamination at all times before and 
during use. Either use them only once or sterilize them in between uses, so each sample 
will be taken by fresh and sterile utensils. 
6.  The type of sample containers used (e.g., bags, tubes, cups, flasks) should depend on the 
type of sample collected. You should use containers that are dry, leak-proof, wide-
mouthed, and of a size suitable for the type of sample collected. 
  The container should be properly labeled, such as with a marked strip of masking 
tape, prior to sampling to identify the sample information, such as the sample 
production batch, time, and the date of sampling. The container should be opened 
only sufficiently to collect the sample directly in the container, and then immediately 
closed and sealed. 
  Containers such as plastic jars or metal cans that are leak-proof may be sealed. 
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  If collecting samples in a container with a lid, the lid should not be placed on a 
counter. 
  Whenever possible, avoid using glass containers, which may break and contaminate 
the sample, the source from which the sample was taken, and/or covered produce, 
including sprouts. 
7.  Sample containers for water samples (i.e., agricultural water and spent sprout irrigation 
water) should not be overfilled, and you should leave an air space of 1 to 2 inches at the 
top to prevent overflow. 
8.  For samples collected indoors, samples and sampling equipment should not be exposed to 
unfiltered air currents. When opening sterile sampling containers, you should work 
rapidly, open sterile sampling containers only to admit the sample and close it 
immediately. 
9.  You should not touch the inside of the sterile sample container, lip or lid. You should not 
allow fingers or anything except the sample to contact the inside of the sample container. 
10. You should not use a sample container that has fallen on the floor. 
11. You should not expose covered produce, including sprouts, or food contact surfaces to 
samples or hands after sampling. You must wash hands with soap and water following 
sample collection, because during sampling hands may have become contaminated in a 
manner reasonably likely to lead to contamination of covered produce ( 
112.32(b)(3)(vi)). 
12. Samples should be delivered to the laboratory promptly. You should ship the samples so 
that they are received by the laboratory within 24 hours from the time of sample 
collection and analyzed promptly. A delay of more than 24 hours between sample 
collection and the labs receipt may make the test results inaccurate (Ref. 98). 
13. Prior to and during delivery or shipment to a laboratory, you should hold your samples 
between 0 and 4.4 oC (between 32 and 40 oF). You should use sealed coolant packs in 
lieu of ice, as needed during delivery or shipment, to avoid the possibility of melting ice 
contaminating the sample. You should not freeze the samples. 
Appendix 2. Collection Procedures  Environmental Sampling 
The two most common methods to collect environmental samples to test for Listeria spp. or 
Listeria monocytogenes are surface sponging and swabbing. Another sampling method used 
is the rinse method. In general, the preferred method of sampling is surface sponging, but 
certain areas could be more appropriately sampled using a swab (e.g., head screws, small water 
collection points, screw holes, threaded surfaces or interior corners of equipment) or rinse 
method. For wet surfaces, you should wipe and absorb moisture and wet product and residue 
with a sponge. For dry surfaces, you should wipe the sample site area with a sponge or swab 
moistened with Dey-Engley (D/E) broth. The sample size of each site should be consistent with 
the testing methodology and the sample collection method being used. The recommended sample 
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size for swabs is generally smaller (e.g., 1 inch by 1 inch) compared to sponges (e.g., 4 inches by 
4 inches up to 12 inches by 12 inches). However, if collecting from surfaces with visible residue 
(e.g., dust, dirt, buildup of organic material), we recommend increasing the number of sponges 
and collecting from a smaller surface area for each sponge to improve the likelihood of detection. 
If Sponge Sampling: 
1.  You should wash and sanitize hands to the mid-forearm before you begin sampling. 
Aseptically place a glove on the hand used for swabbing. 
2.  Using the ungloved hand, open the bag containing the sponge on a stick by pulling off the 
clear perforated strip at the top of the bag. 
3.  Pull apart the white tabs to open the mouth of the bag. 
4.  If not using pre-moistened sponges, aseptically pour 9-10 ml of sterile D/E or other 
neutralizing broth into the bag to hydrate the sponge, being careful not to contaminate the 
broth or sponge during the transfer. 
5.  Close the bag and evenly moisten sponges by hand massage. 
6.  Position the sponge so that the handle is sticking out of the bag and close the bag around 
the stick. 
7.  Through the bag, squeeze the excess broth gently out of the sponge. Do not let your hand 
go past the thumb stop on the stick. 
8.  Carefully take the sponge-stick out of the bag by grasping the stick and swab the area 
selected using firm and even pressure. Be careful to maintain sanitary conditions when 
sampling. Do not let your hand go past the thumb stop on the handle. 
9.  Swab the chosen area (e.g., between 4 inches by 4 inches up to 12 inches by 12 inches 
square of food contact or environmental surface area): 
  Sponge vertically (approximately 10 times); then 
  Flip the sponge and use the other side to swab horizontally (approximately 10 times); 
  Then move the sponge diagonally, using the same surface side as you used for 
horizontal (approximately 10 times). 
10. Open the bag and insert the sponge portion into the bag. 
11. Grip the sponge through the bag and bend the stick of the sponge back and forth with 
slight force, while gripping the sponge through the bag. The stick should break easily 
within the sponge. (Do not break the stick at the thumb stop.) Discard the broken stick. If 
the stick is sticking out above the sponge, discard this sample. 
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12. Squeeze as much air out of the bag as possible and fold the top of the bag down at least 3 
times until it is folded all the way down to the sponge. Fold in the tabs to lock the fold in 
place. 
13. Label the bag with the date and location of the sample. 
14. Take each new sample following the same steps in 1  13, starting by changing the glove 
on the gloved hand between samples. 
15. Prior to and during delivery or shipment to a laboratory, samples should be kept 
refrigerated. Sealed coolant packs should be used in lieu of ice, as needed during delivery 
or shipment, to avoid the possibility of melting ice contaminating the sample. Samples 
should not be frozen. Samples should be shipped so that they are received by the 
laboratory within 24 hours after sample collection. We recommend that the maximum 
timeframe between environmental sampling and analysis be 48 hours (Ref. 94).  
If Swab Sampling: 
1.  You should wash and sanitize hands to the mid-forearm before you begin sampling. 
Aseptically place a glove on the hand used for swabbing. 
2.  Using the ungloved hand, open the tube containing the swab. 
3.  If not using pre-moistened swabs, aseptically pour 9-10 ml of sterile D/E or other 
neutralizing broth into the tube to hydrate the swab or use sampling swabs that are 
already pre-moistened in 9-10 ml of D/E broth. If the D/E broth is not purple, discard the 
tube. 
4.  Close the cap of the tube and wait until the swab is moistened. 
5.  Carefully take the swab out of the tube and swab the area selected using firm and even 
pressure. Be careful to maintain sanitary conditions when sampling. 
6.  Swab the chosen area (e.g., 1 inch by 1inch square of food contact or environmental 
surface area, or insert the swab into the crack, crevice or hole) using firm and even 
pressure: 
  Swab the 1 inch by 1inch surface vertically (approximately 10 times); then 
  Rotate the swab and use the other side to swab horizontally (approximately 10 times); 
then 
  Move the swab diagonally, using the same surface side as you used for horizontal 
(approximately 10 times).  
  For cracks, crevices and holes, rotate the swab in the area as you move it around the 
area. 
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7.  Do not touch the outside of the opening of the tube; insert the swab portion into the tube. 
8.  Close the cap of the tube. 
9.  Label the bag with the date and location of the sample. 
10. Take each new sample following the same steps in 1  9, starting by changing the glove 
on the gloved hand between samples. 
11. Prior to and during delivery or shipment to a laboratory, samples should be kept 
refrigerated. Sealed coolant packs should be used in lieu of ice, as needed during delivery 
or shipment, to avoid the possibility of melting ice contaminating the sample. Samples 
should not be frozen. Samples should be shipped so that they are received by the 
laboratory within 24 hours after sample collection. We recommend that the maximum 
timeframe between environmental sampling and analysis be 48 hours (Ref. 94).  
Collecting Rinse Samples 
To collect samples using a rinse technique, you should add small pieces from equipment (such as 
screws, nuts or gaskets) directly to the bag containing D/E broth and hand massage the bag for 
sufficient time to remove soil and residues (approximately 1 minute). Then aseptically remove 
the items from the bag and subject the broth to analysis. 
In some situations, involving small cracks and crevices, it may help to use a plastic bulb transfer 
pipette. You should use tubes containing 10 mL sterile D/E broth in this procedure. You should 
pull the D/E broth into the pipette bulb and transfer the D/E broth to the crack or crevice, and 
then you should pull it back into the bulb. You should repeat this several times to thoroughly 
rinse the crack or crevice.  
Collecting Liquid Samples (Including Floor Drain Effluents) 
We recommend that you use a sterile plastic beaker or similar container to collect 110 + 5 ml of 
liquids, where possible, such as drainage effluents, standing water, melt water from thawed 
processing ice, and vacuum or drip pan condensate. We recommend that you immediately 
transfer the collected sample into a sterile screw-capped bottle and then chill and store the bottle 
at 5 degrees C (41 degrees F), including during transport to the testing laboratory. 
Preparing Samples Collected from Liquids 
For larger samples (e.g., 100 mL or greater), we recommend that you filter 100 ml of the 
collected liquid through one or more sterile 0.45 micron pore-diameter filters as soon as possible 
after sample collection. If particulate content is high (e.g., judging from the sample turbidity), we 
recommend that you pass the liquid through a sterile glass pre-filter before the 0.45 micron filter. 
You should rinse a retained substance on the filter plus any pre-filter with 5-10 ml of D/E broth 
to remove any residual inhibitory substances. If necessary, you should cut the filters from the 
funnel devices, using sterile scalpels. You should put each filter and the pre-filter, if any, in a 
sterile bag (if you will use a Stomacher) or in a sterile container (such as a blender jar, if you use 
a blender). You should add 225 mL of UVM broth, and follow procedures in Testing 
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Methodology for Listeria species of L. monocytogenes in Environmental Samples (version 1, 
Oct 2015) (currently available at: https://www.fda.gov/media/94358/download) beginning with 
incubation of the primary enrichment. 
For small volumes of liquid samples, we recommend that you add the liquid sample to 225 mL 
of UVM broth and follow procedures in Testing Methodology for Listeria species of L. 
monocytogenes in Environmental Samples (version 1, Oct 2015) beginning with incubation of 
the primary enrichment. 
Appendix 3. Potential Sources of L. monocytogenes for Sampling in a Sprout 
Operation 
This table provides examples of possible food contact and non-food contact surfaces for use in 
developing a Listeria environmental monitoring program. The list is not all-inclusive and is not 
intended to be determinative; rather, we provide the examples below as an aid. We recommend 
that firms develop their own robust assessments of the food contact and non-food contact 
surfaces in their operations, as well as an understanding of appropriate zone designations. 
Table 7. Potential Sources of L. monocytogenes in a Sprout Operation10 
Sample Location  Zone 1  Zone 2  Zone 3  Zone 4  Notes 
   
Aprons  X  X  Aprons reasonably likely to 
contact sprouts would be 
considered Zone 1. 
   
Conveyors  X  X  Fibrous and porous-type 
conveyor belts, hollow rollers for 
conveyances are of particular 
concern. 
     
Ceilings and shields  X  For the purpose of your 
from which condensate  environmental monitoring 
or drainage can drip  program, we recommend you 
onto the product  focus on the surfaces that 
directly contact the food  
Ceilings that are not    X  X    Condensate drip pans are of 
above sprouts,  particular concern. 
overhead structures, 
and catwalks 
 
10 The table has been added to the document to facilitate a better understanding of food contact surfaces (FCSs) and 
non-FCSs, and zone designations.  The contents are based on information that was available in the draft Guidance 
for Industry: Compliance with and Recommendations for Implementation of the Standards for the Growing, 
Harvesting, Packing, and Holding of Produce for Human Consumption for Sprout Operations (Ref. 99), and are 
now presented in a more user-friendly format. 
91 
  Contains Nonbinding Recommendations 
Sample Location  Zone 1  Zone 2  Zone 3  Zone 4  Notes 
   
Gloves  X  X  Gloves that are reasonably likely 
to contact sprouts would be 
considered Zone 1. 
       
Growing bins or trays  X 
       
Rotary drums  X 
       
Spin dryer (for  X 
sprouts) 
     
Tools for cleaning  X  e.g., Brushes and scouring pads, 
equipment  cracked hoses 
       
Utensils  X 
   
Work table  X  X  The surface that is reasonably 
likely to contact sprouts is Zone 
1; the underside and supports are 
Zone 2. 
 
Walls  X  X  X  Zone depends on proximity to 
food contact surfaces and food. 
   
Equipment housing or  X  X  Wet, rusting, or hollow 
framework  framework, open bearings within 
equipment, motor housings, 
on/off switches are areas of 
particular concern. 
     
Pallets  X  X 
 
Wash areas   X  X  X  e.g., Washing tubs, sinks 
   
Maintenance tools  X  X  e.g., Wrenches and screw drivers 
   
Coolers  X  X  Wet insulation in walls or around 
pipes and cooling units, rubber 
seals around doors are of 
particular concern. 
       
Floors and drains in  X 
the immediate vicinity 
of FCSs 
92 
  Contains Nonbinding Recommendations 
Sample Location  Zone 1  Zone 2  Zone 3  Zone 4  Notes 
Floors or drains in or      X     
near the production 
areas but not in the 
immediate vicinity of 
FCSs 
       
Racks for trays  X 
       
Surfaces employees  X 
handle prior to 
handling the product 
     
Forklifts  X  X 
     
Vacuum cleaners and  X  X 
floor scrubbers 
     
Trash cans   X  X 
       
Hand trucks and carts  X 
that move in the 
production area 
       
Cafeterias  X 
       
Hallways outside the  X 
production area 
       
Locker rooms  X 
   
Areas outside of the  X  X  X 
production area in 
which raw materials 
(e.g., seeds) or finished 
products are stored or 
transported 
 
   
93 
  Contains Nonbinding Recommendations 
IX. Paperwork Reduction Act of 1995 
This guidance contains information collection provisions that are subject to review by the Office 
of Management and Budget (OMB) under the Paperwork Reduction Act of 1995 (44 U.S.C. 
3501-3521). 
 
The time required to complete this information collection is estimated to average 195 minutes 
per response, including the time to review instructions, search existing data sources, gather the 
data needed, and complete and review the information collection.  Send comments regarding this 
burden estimate or suggestions for reducing this burden to: 
 
Food and Drug Administration  
Center for Food Safety and Applied Nutrition 
Office of Food Safety 
5001 Campus Drive 
College Park, MD 20740 
(Tel) 240-402-1700 
 
 
An Agency may not conduct or sponsor, and a person is not required to respond to, a collection 
of information unless it displays a currently valid OMB control number. The OMB control 
number for this information collection is 0910-0816. To find the current expiration date, search 
for this OMB control number at https://www.reginfo.gov.  
 
 
94 
  Contains Nonbinding Recommendations 
X. References 
The following references marked with an asterisk (*) are on display at the Dockets Management 
Staff (HFA-305), Food and Drug Administration, 5630 Fishers Lane, Rm. 1061, Rockville, MD 
20852, 240-402-7500, and are available for viewing by interested persons between 9 a.m. and 4 
p.m., Monday through Friday; they also are available electronically at 
https://www.regulations.gov. References without asterisks are not on public display at 
https://www.regulations.gov because they have copyright restriction. Some may be available at 
the website address, if listed. References without asterisks are available for viewing only at the 
Dockets Management Staff. FDA has verified the website addresses, as of the date this document 
publishes in the Federal Register, but websites are subject to change over time. 
 
 
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95 
  Contains Nonbinding Recommendations 
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96 
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