FSIS Ready-to-Eat Fermented, Salt-

Cured, and Dried Products Guideline

May 5, 2023

Document ID: FSIS-GD-2023-0002

calicersim

This guideline provides information

on the Agency regulatory
requirements associated with safe
production of ready-to-eat (RTE)
shelf-stable, fermented, salt-cured,
and dried products that rely on
multi-hurdle approaches to achieve
lethality and shelf-stability. It
applies to small and very small
meat and poultry official
establishments although large
establishments can also benefit
from the information. It relates to 9
CFR part 417.

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 Table of Contents

Preface .......................................................................................................................................................... 4
Purpose ..................................................................................................................................................... 4
Reason for Issuing the Guideline .............................................................................................................. 5
How to Effectively Use this Guideline ........................................................................................................... 6
Questions Regarding Topics in this Guideline............................................................................................... 7
Background ................................................................................................................................................... 8
Products Covered by this Guideline .......................................................................................................... 8
Products NOT covered by this Guideline .................................................................................................. 9
Biological Hazards of Concern in Multi-hurdle Lethality Products ........................................................... 9
Recommended Targets for Biological Hazards ....................................................................................... 10
Steps or Hurdles to Ensure Food Safety.................................................................................................. 10
Table 1. Key Steps or Hurdles Used to Achieve Lethality and Shelf-Stability by Product Group ........ 11
Validation – Element 1: Scientific Support ............................................................................................. 11
Table 2. Example of Side-by-Side Comparison of Parameters Used in the Support vs. the Actual
Process and Rationale for Differences ................................................................................................ 12
CASE STUDY: The importance of using critical operational parameters consistent with the scientific
support ................................................................................................................................................ 13
Validation – Element 2: In-plant validation ............................................................................................ 13
Overview of Fermented Products ............................................................................................................... 15
Table 3. Overview of hazards of concern establishments should consider in the hazard analysis
during lethality and stabilization and typical controls for fermented products. ................................ 15
Overview of Salt-Cured Products ................................................................................................................ 18
Table 4. Overview of hazards of concern establishments should consider in the hazard analysis
during lethality and stabilization and typical controls for salt-cured products. ................................. 18
Overview of Dried Products ........................................................................................................................ 20
Table 5. Overview of hazards of concern establishments should consider in the hazard analysis
during lethality and stabilization and typical controls for dried products.......................................... 20
Post-lethality Considerations ...................................................................................................................... 22
References .................................................................................................................................................. 24
Appendix 1: Labeling Considerations for Not Ready-to-Eat (NRTE) fermented, salt-cured, and dried
products. ..................................................................................................................................................... 33
Appendix 2: Biological Hazards of Public Health Concern for RTE Shelf-Stable Fermented, Salt-Cured, and
Dried Products ............................................................................................................................................ 35

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 Table 6. Salmonella and E. coli O157:H7 Foodborne Illness Outbreak History in Fermented and Salt-
Cured, and Dried Ready-to-Eat Meat Products Produced in the U.S. ................................................ 35
What About Mold? ............................................................................................................................. 39
Appendix 3: Lethality and Shelf-Stability Targets ...................................................................................... 40
Lethality Targets (Salmonella, STEC, and Lm) ......................................................................................... 40
What Options do I have if my Steps/Hurdles don’t Achieve a 5.0-log Reduction? ................................ 41
How Can I Apply the Concept of Raw Batter Testing to Whole Muscle Products? ................................ 43
Shelf-Stability Targets ............................................................................................................................. 44
Can I Use Finished Product Testing Alone if I Don’t have Scientific Support for my Process? ............... 44
Appendix 4: Considerations for Different Types of Scientific Support ....................................................... 45
Appendix 5: Scientific Support Available for Shelf-Stability ...................................................................... 48
Appendix 6: Critical Operational Parameters for Fermentation ................................................................ 49
Why is Using a Starter Culture Important? ............................................................................................. 51
Appendix 7: Fermentation Deviations ........................................................................................................ 57
Appendix 8: Critical Operational Parameters for a Low-Temperature Heat Step ..................................... 60
Appendix 9: Critical Operational Parameters for Salt-Curing and Equalization ........................................ 62
Appendix 10: Critical Operational Parameters for Seasoning/Marination of Dried Products ................... 67
Appendix 11: Critical Operational Parameters for Drying ......................................................................... 68
Appendix 12: Scientific Support Available for Lethality in Dry and Semi-Dry Fermented Sausages ......... 73
Table 7. Summary of Scientific Support Available for Lethality in Dry and Semi-Dry Fermented
Sausages .............................................................................................................................................. 73
Appendix 13. Scientific Support Available for Lethality in Salt-Cured Products ........................................ 82
Table 8. Summary of Scientific Support Available for Lethality in Basturma. ................................... 82
Table 9. Summary of Scientific Support Available for Lethality in Country Cured Ham. ................... 85
Table 10. Summary of Scientific Support Available for Lethality in Bresaola. ................................... 86
Appendix 14: Scientific Support Available for Lethality in Dried Products ................................................. 87
Table 11. Summary of Scientific Support Available for Lethality in Droëwers. ................................. 87
Table 12. Summary of Scientific Support Available for Lethality in Biltong....................................... 88
Appendix 15: Designing Challenge Studies for Fermented, Salt-Cured, and Dried Products ..................... 95
Table 13. Potential surrogates for lethality challenge studies conducted in-plant. .......................... 97
Appendix 16: Glossary .............................................................................................................................. 101

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Preface

This guideline represents FSIS’ current thinking on this topic and should be considered
usable as of this issuance. The information in this guideline is provided to assist meat
and poultry establishments in meeting the regulatory requirements. The contents of this
document do not have the force and effect of law and are not meant to bind the public in
any way. This document is intended only to provide clarity to industry regarding existing
requirements under the regulations. Under the regulations, meat and poultry
establishments may choose to implement different procedures than those outlined in
this guideline, but they would need to validate and support how those procedures are
effective.

This guideline is focused on small and very small plants in support of the Small
Business Administration’s initiative to provide small businesses with compliance
assistance under the Small Business Regulatory Enforcement Fairness Act (SBREFA).
However, all meat and poultry establishments may apply the recommendations in this
guideline. It is important that small and very small establishments have access to a full
range of scientific and technical support, and the assistance needed to establish safe
and effective Hazards Analysis and Critical Control Point (HACCP) systems. Although
large plants can benefit from the information, focusing the guideline on the needs of
small and very small establishments provides them with assistance that may be
otherwise unavailable to them.

Purpose

This guideline is designed to respond to commonly asked questions from small and very
small establishments that manufacture RTE, shelf-stable, fermented, salt-cured, and
dried meat and poultry products where cooking is not the primary lethality step about:

• Recommended targets for reduction or prevention of each hazard.

• The key steps in each process needed to ensure safety.
• The critical operational parameters associated with each step.
• The biological hazards associated with and scientific support available to help
produce the following products:
o Fermented dry and semi-dry fermented sausages including:
 Lebanon bologna
 Summer sausage
 Pepperoni
 Salami, including Genoa
 Soudjouk
o Salt-cured products including:
 Basturma
 Country cured ham
 Bresaola
o Dried products including:
 Biltong
 Droëwors.

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Establishments can always seek guidance from State university extension service
specialists and HACCP Coordinators on developing food safety plans not provided in
this guideline to comply with HACCP regulatory requirements. Small and very small
establishments can also seek guidance from the Niche Meat Processor Assistance
Network, a university Extension-based community of practice.

Reason for Issuing the Guideline

FSIS has several documents that address the safe production of ready-to-eat (RTE)
meat and poultry products. However, based on questions received, FSIS determined
that the current documents listed below do not adequately address the specific
considerations related to supporting the lethality and shelf-stability of RTE shelf-stable
fermented, salt-cured, and dried meat and poultry products. The available documents
for RTE products include:

• The FSIS Cooking Guideline for Meat and Poultry

Products (Revised Appendix A) - provides guidance on
using cooking as a lethality treatment.
• The FSIS Stabilization Compliance Guideline for Meat
and Poultry Products (Revised Appendix B) - provides KEY DEFINITIONS
guidance on stabilization treatments (primarily cooling
and hot-holding).
• The FSIS Compliance Guideline for Meat and Poultry Ready-to-eat meat and
Jerky Produced by Small and Very Small Establishments poultry is defined by FSIS in
- provides guidance about how to safely produce jerky. 9 CFR 430.1 as a meat or
• The FSIS Guideline: Controlling Listeria monocytogenes poultry product that is in a
in Post-Lethality Exposed RTE Meat and Poultry form that is edible without
Products - provides guidance on complying with the additional preparation to
requirements for 9 CFR 430 to address Listeria achieve food safety and may
monocytogenes (Lm) contamination in the post-lethality receive additional
exposed environment. preparation for palatability or
• FSIS Guideline for the Prevention and Control of aesthetic, epicurean,
Trichinella and Other Parasitic Hazards in Pork and gastronomic, or culinary
Products Containing Pork - provides guidance on purposes.
understanding the available options that are effective for
the prevention and control of Trichinella spiralis and other Shelf-stable for the
parasitic hazards in pork. purposes of meat and poultry
products is defined as the
FSIS had some information related to the safe production of condition achieved when
fermented, salt-cured, and dried meat and poultry products that meat and poultry products
was previously available in the following documents and has can be stored under ambient
been now incorporated into this guideline: temperature and humidity
conditions; if the package
• “Food Safety Lessons Learned from the Lebanon integrity is maintained during
Bologna Outbreak” – Although Lebanon bologna is a storage, shipping, and
unique product which is fermented to a low pH, FSIS display at retail and in the
incorporated information from that guideline into this home; and the product will
not spoil or become unsafe
throughout the
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manufacturer’s specified
shelf-life.
 larger document because a lot of the information can be applied to other semi-
dry fermented products (Getty, 1999; Vignolo et al., 2010).
• “Challenge Study – Escherichia coli O157:H7 in Fermented Sausage” – relevant
information was incorporated into Appendix 15 of this document.

The guideline also includes lessons learned from two salmonella outbreaks in 2021
associated with ready-to-eat fermented, dried, and salt-cured meat Italian-style meat
products. These lessons learned are also included in FSIS’ Outbreak Investigation After
Action Review.

How to Effectively Use this Guideline

This guideline is organized to provide users with an overview of topics related to the
safe production of RTE shelf-stable fermented, salt-cured, and dried meat and poultry
products. Additional details about each topic are included in appendices. To use this
guideline, FSIS recommends that readers review the overview of each of the topics and
consult relevant appendices for more detail. Hyperlinks will quickly take you to the
correct place in the document electronically and are also provided to other
complementary guidelines. Users should note that words that are bolded throughout
the document are defined in the Glossary in Appendix 16.

Appendices 12, 13, and 14, contain summaries of available scientific articles. These
summaries are provided to help establishments identify relevant scientific support for
supporting decisions in the hazard analysis. Establishments are not limited to using
these articles as support, and the summaries are not adequate support on their own
because they do not contain the details of each study and the establishment needs to
determine if it is representative of the actual process. For this reason, establishments
will need to have the full copy of the article on-file. Links to full copies of articles have
been provided in the References section when available.

How to Comment on the Guideline

FSIS is seeking public comment on this guideline as part of its efforts to continuously
assess and improve the effectiveness of policy documents. All interested persons may
submit comments regarding any aspect of this document, including but not limited to
content, readability, applicability, and accessibility. The comment period will be 60 days
from publication of the Federal Register Notice and, as appropriate, the agency may
update this guideline in response to comments. Although FSIS may make changes to
the guideline in response to comments, this document reflects FSIS’s current thinking,
and FSIS encourages establishments producing products discussed in this document to
review it.

Comments may be submitted by either of the following methods:

• Federal eRulemaking Portal Online submission at regulations.gov. This website

provides a way to type short comments directly into the comment field on the
webpage or attach a file to submit lengthier comments. Follow the online
instructions at that site to submit comments.
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 • Mail and hand- or courier-delivered items: Send to Docket Clerk, U.S.
Department of Agriculture (USDA), FSIS, 1400 Independence Avenue SW,
Room 6065, Washington, D.C. 20250-3700.

All items submitted by mail or electronic mail must include the Agency name, FSIS, and
document title: FSIS Ready-to-Eat Fermented, Salt-Cured, and Dried Products
Guideline. Comments received will be made available for public inspection and posted
without change, including any personal information, to http://www.regulations.gov.

Questions Regarding Topics in this Guideline

If after reading this guideline you still have questions, FSIS recommends searching the
publicly posted Knowledge Articles (“Public Q&As”) in the askFSIS database. If after
searching the database, you still have questions, refer them to the Office of Policy and
Program Development through askFSIS and select HACCP Deviation & HACCP
Validation as the Inquiry Type or by telephone at 1-800-233-3935.

Documenting these questions helps FSIS improve and refine present and future
versions of the guideline and associated issuances.

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 FSIS RTE Fermented, Salt-Cured, and Dried Products
Guideline

Background

Multi-hurdle products in the context of this guideline are those products that rely on a
combination of hurdles or processing steps to eliminate pathogens of concern and result
in a RTE shelf-stable product. These hurdles typically include processing steps such as
fermentation, salt-curing, and drying that use a combination of factors such as reduced
pH, reduced water activity (also referred to as aw) over time, high brine, or salt
concentration to kill bacteria and prevent their outgrowth during storage. Generally, one
step is not sufficient on its own to eliminate pathogens of concern, instead a
combination of processing steps or hurdles is needed.

Products Covered by this Guideline

This guideline focuses on the safe production and supporting documentation for the
following RTE shelf-stable products that are produced under the Not Heat-Treated
Shelf-Stable HACCP category or Heat-Treated Shelf-Stable HACCP category.
Examples of scientific support are included in appendices for products in bold because
these are common products FSIS receives questions about and are products where
scientific support is available:

Product Products in this Category Include… For More

Group Information
(Go to page):
Fermented Genoa salami, hard salami, pepperoni, turkey 15
pepperoni, summer sausage, Abruzzese, Lebanon
bologna, sopressata, thuringer, mettwurst, saucisson,
chorizo, chourico, soudjouk (sujuk or soujouk), pickled
pigs’ feet, bologna in vinegar, landjager
Salt-cured Prosciutto ham, Parma ham, Westphalian ham, 18
Bayonne ham, Serrano ham, Black Forest ham, country
ham, pancetta, coppa, capocolla, bresaola, beef
prosciutto, basturma, duck prosciutto, linguica,
salchichon
Dried dried beef, beef jerky 1, beef nuggets, steak tenders, 20
kippered beef, meat sticks, turkey jerky, tasajo,
pemmican, pipi kaula, droëwors, biltong, jamon
(jambon), longanisa, (some) saucisson, (some) chorizo,
dried soup mixes/soup bases, freeze-dried entrees, fried
pork skins/rinds/cracklings/chicharrones, lard

1
Jerky is not covered in this guideline as explained on the next page.
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Products NOT covered by this Guideline

This guideline does not address the production of products

classified as not ready-to-eat (NRTE) or not shelf-stable;
however, labeling considerations are provided in Appendix KEY DEFINITIONS
1 to address commonly asked questions.

This guideline does not cover jerky, which is considered a Water activity, abbreviated
dried product, as most jerky processes rely on cooking as aw, is a measure of the
alone (e.g., by following the FSIS Cooking Guideline for concentration of moisture
Meat and Poultry Products (Revised Appendix A)) to (i.e., water) and its
achieve lethality. Guidance regarding the production of jerky availability in a food. The
can be found in the FSIS Guideline for Meat and Poultry amount of water available in
Jerky Produced by Small and Very Small Establishments. a food depends on the total
concentration of all dissolved
Biological Hazards of Concern in Multi-hurdle substances in the product
Lethality Products because they bind water.
Thus, if ingredients such as
The following section and the supplemental information in salt or sugar are added to
Appendix 2 are designed to complement FSIS’ Meat and food, they compete with the
Poultry Hazards and Control Guide and to further assist bacteria for available water.
establishments in conducting a hazard analysis for
fermented, salt-cured, and dried products as required by 9
CFR 417.2(a)(1) and for supporting decisions in the hazard analysis as required by 9
CFR 417.5(a)(1).

Outgrowth of the following are hazards in raw products that fermentation, salt-
curing, or drying steps should be designed to limit in the finished RTE
product:

• Staphylococcus aureus (S. aureus)

• Clostridium perfringens (C. perfringens)

• Clostridium botulinum (C. botulinum)

The following are hazards present in raw products that the fermentation, salt
curing, or drying steps should be designed to destroy in the finished RTE
product:

• Salmonella

• STEC

• Lm

• Trichinella spiralis (T. spiralis) and Toxoplasma gondii (T. gondii) (greater risk
of infection for feral or non-confinement raised swine)

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These hazards may be of concern at various points
in the production process and multiple hurdles may
be needed to address each hazard. Appendix 2 has
some specific considerations that can help inform
an establishment’s hazard analysis decision-
making. Information on mold as a potential
biological hazard of concern can also be found in KEY DEFINITIONS
Appendix 2.

Recommended Targets for Biological Hazards Lethality is the process or

combination of processes that
For each biological hazard identified, ensures that no Salmonella
establishments need to identify targets for reduction organisms remain in the
or prevention. Targets are quantifiable pathogen finished product, as well as
reduction levels or growth limits set by the reduces other pathogens and
establishment to produce safe products in the their toxins or toxin metabolites.
absence of regulatory performance standards. Examples of lethality processes
Targets are used by the establishment to include cooking, fermentation,
demonstrate that the lethality and shelf-stability salt-curing, and drying.
processes achieved by their food safety system
prevents, eliminates, or reduces pathogens to Stabilization is the process of
acceptable levels. For example, FSIS recommends preventing or limiting the growth
that the lethality treatment (i.e., the combination of of spore-forming bacteria
hurdles or steps) for RTE shelf-stable meat and capable of producing toxins
poultry products achieve at least a 5.0-log reduction either in the product before
of Salmonella, STEC (in beef), and at least a 3.0-log consumption or in the human
reduction in Lm. For more information on lethality intestine after consumption.
and shelf-stability targets see Appendix 3. Establishments may use a
variety different stabilization
Steps or Hurdles to Ensure Food Safety processes such as cooling, hot-
holding, or meeting and
For fermented, salt-cured, and dried products, maintaining certain pH or water
lethality and shelf-stability are achieved using activity levels.
multiple process steps or hurdles. The following are
the key process steps and hurdles used to achieve
lethality and shelf-stability in fermented, salt-cured, and dried products. The key hurdles
and critical operational parameters for each will be described in more detail for each
process.

NOTE: The processing steps used to achieve shelf-stability also often stabilize a
product to prevent or limit the growth of spore-forming bacteria by reducing the pH or
water activity. For more information on stabilization see FSIS’ Stabilization Guideline for
Meat and Poultry Products (Revised Appendix B).

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 Table 1. Key Steps or Hurdles Used to Achieve Lethality and Shelf-Stability by Product Group
Seasoning/ Fermentation Low-Temperature Salt-curing/ Drying
Marination Heat Step Equalization
Hurdle/ Addition of salt Lower pH/ Heat High brine/Reduced Reduced
Product and nitrite or Competitive water activity water
Group nitrate/Lower pH Microflora/ activity
Bacteriocin over time
production
Fermented X X Optional X

Salt-cured X Optional X X

Dried X X

In addition to applying multiple hurdles, it is also important that establishments

understand and:

• Adhere to the basic principles of Sanitation Standard Operating Procedures

(SSOPs) (9 CFR 416).
• Follow Good Manufacturing Practices (GMPs).
• Address product handling post-lethality.

Validation – Element 1: Scientific Support

Once an establishment identifies the key steps in its process it should identify the
scientific support available that closely matches the actual process and shows that the
process achieves the desired lethality target as part of the validation process. There are
two distinct elements to validation: 1) the scientific or technical support for the HACCP
system design (design) and 2) the in-plant validation data (execution). This guideline
focuses on the scientific support available to meet the first element of validation. As
discussed in the FSIS HACCP Systems Validation Guideline, to meet the first element
of validation (i.e., the scientific or technical support) establishments may use:

• Published processing guidelines.

• Peer-reviewed scientific or technical data or information.
• Expert advice from processing authorities.
• Validated pathogen modeling programs.
• Challenge or inoculated pack studies.
• Data gathered by the establishment in-plant.
• Regulatory performance standards.
• Best practice guidelines.

Any of these types of support may be acceptable provided they are complete and the
critical operational parameters match the establishment’s process. Some specific
considerations for each of the types of supporting documents when used to support
lethality and shelf-stability of fermented, salt-cured, and dried products are given in
Appendix 4.

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 Establishments should carefully identify the critical operational parameters used in
the actual process and compare those to the ones used in the scientific support. Critical
operational parameters are the specific conditions that the intervention must operate
under for it to be effective. Examples of critical operational parameters used during
fermentation, salt-curing, and drying include:

• Antimicrobial application.
• Fermentation temperature, target pH, time to reach target pH, and smoke (if
used).
• Type and use of starter cultures.
• Curing time.
• Curing temperature.
• Salt coverage of exposed muscle tissue.
• Drying room temperature.
• Drying time (i.e., days or weeks).
• Product characteristics including product formulation.

Establishments must implement critical operational parameters in the actual production

process consistent with the parameters in the scientific or technical support or provide
justification for why differences would not impact the efficacy of the intervention (9 CFR
417.5(a)(1)). When different levels of a critical operational parameter other than those in
the support document are used, establishments should consider developing a decision-
making document that explains the scientific rationale for why the different level would
not affect the efficacy of the intervention or process or why the different studies support
the effectiveness of the process when combined. This scientific rationale should include
reference to data or scientific principles as support. Establishments may be able to use
in-plant validation data when differences between the actual process and support are
small. For more information see the Appendix 4 section about “Data gathered by the
establishment in-plant”. FSIS recommends establishments consider making a table to
do a side-by-side comparison of the parameters in the actual process and those used in
the actual study. An example is provided below in Table 2.
Table 2. Example of Side-by-Side Comparison of Parameters Used in the Support vs. the
Actual Process and Rationale for Differences
Process Critical Level Used in Level Used in Rationale for Why the
Step Operational the Scientific Establishment’s Difference is
Parameter Support Actual Process Acceptable
Post- Hold time and Internal Internal Actual process uses
fermentation temperature temperature of temperature of higher temperature and
low 128°F for 1h 134°F for 1h same dwell time which
temperature from Hinkens et should result in same
heat step al., 1996 or higher reductions
per FSIS Cooking
Guideline for Meat and
Poultry Products
(Revised Appendix A).

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CASE STUDY: The importance of using critical operational parameters consistent with the scientific
support
• In March 2011, there was a recall of a Lebanon bologna product that was associated with a
foodborne illness outbreak of E. coli O157:H7.
• FSIS’ investigation revealed the establishment had not properly validated their process. The
establishment’s supporting documentation for the critical operational parameters did not match
the actual commercial process used:

Critical operational Actual process Supporting Documentation

parameter
Diameter 52 to 119 mm 27 mm
Casing Permeable casing Impermeable sealed glass
tube
Cooking equipment Large smokehouse fitted with Well-controlled water bath
single heat source, humidity
was not well-controlled

• Differences likely led to a lower reduction in foodborne pathogens of concern in the actual
process than what was demonstrated in the supporting documentation.
• This outbreak highlights the importance of identifying supporting documentation that is
representative of the actual process so that the results can be repeatable.

Validation – Element 2: In-plant validation

As discussed in the FSIS HACCP Systems Validation Guideline, if establishments have

adequate scientific support and implement the critical operational parameters in the
actual process consistent with the scientific support then to meet the second element of
validation (i.e., the scientific or technical support) the establishment should collect in-
plant data that demonstrates that the critical operational parameters are being met.
Establishments should develop the appropriate in-plant data during the initial 90 days of
implementing a new HACCP system, or whenever a new or modified food safety hazard
control is introduced into an existing HACCP system (e.g., as implemented after a
HACCP plan reassessment).

NOTE: For some salt-cured products, the initial validation period may extend beyond 90
calendar days due to the nature of the process and the length of time it takes to
implement the critical operational parameters that impact lethality. To determine
whether the system is properly designed and executed, even though the regulations
provide 90 days for initial validation, an establishment needing more than 90 days can
ask the district office, in writing, for additional time to collect at least 13 production days
of records when it first starts operating, when it begins producing new product, or for a
modified HACCP plan if the results of a reassessment indicate additional support is
needed. In the request, an establishment should state why more than 90 days are
needed to collect the in-plant validation data, and how it plans to gather at least 13
production days’ worth of in-plant validation data. The request will then be evaluated on
a case-by-case basis. The establishment should consider focusing validation activities
13
on the product produced most frequently within each HACCP category. In addition, the
establishment may consider evaluating data collected for products across multiple
HACCP categories to determine whether the data together can support its ability to
meet critical operational parameters (80 FR 27557).

A discussion of how microbiological data gathered as part of initial validation can be

used to support a process when differences between the scientific support and the
actual process are small can be found in Appendix 4 under the section about “Data
gathered by the establishment in-plant”.

The next part of the guideline includes an overview of each of the three following types
of products: 1) fermented, 2) salt-cured, and 3) dried products:

• The biological hazards of concern for fermented, salt-cured, and dried products.

• The recommended targets to address those biological hazards.

• The steps or hurdles to ensure food safety.

• The types of scientific support that can support the recommended targets are
met.

• The importance of understanding the critical operational parameters in your

process and how they relate to those used in the scientific support.

14
 Overview of Fermented Products
RTE fermented meat and poultry products are products in which the raw meat or poultry
are usually reduced in size by grinding or chopping, formulated with cure, starter
culture, salt and seasoning mixture, stuffed in casings, fermented, sometimes heated
with a low temperature heat step for food safety or smoked, and then dried. There are
also RTE acidified products that are formulated with chemical acidulants, instead of
starter cultures, to accelerate the acidification process by eliminating the lengthy
fermentation step. This section focuses on fermented products. Products in this
category include: Genoa salami, hard salami, pepperoni, turkey pepperoni,
summer sausage, Abruzzese, Lebanon bologna, sopressata, thuringer, mettwurst,
saucisson, chorizo, chourico, soudjouk (sujuk or soujouk), pickled pigs’ feet, bologna
in vinegar, and landjager. See examples of scientific support for products in bold in
Appendix 12.
Table 3. Overview of hazards of concern establishments should consider in the hazard
analysis during lethality and stabilization and typical controls for fermented products.
Hazard Source Adulterant Recommended Fermentation Drying
(Yes/No) Target
Salmonella Raw meat Yes – 5-log reduction Effectiveness for Effectiveness for
STEC (beef) and zero for Salmonella Salmonella, STEC, and Salmonella, STEC,
Lm poultry, tolerance and STEC Lm depends on: and Lm depends on:
spices, 3-log reduction Fermentation Drying room
herbs for Lm temperature, target pH, temperature
and time to reach target Drying time
For more pH; Target water activity
information Starter culture; Product characteristics
including on Product characteristics; See Appendix 11
alternative Consider use of a low
lethality targets temperature heat step
see Appendix 3 See Appendix 6
S. aureus Raw meat Yes – During For S. aureus, degree- Drying is effective for
and depending production: no hours concept during S. aureus during
poultry on level more than 2-log production storage after water
spices, growth; See page 37 and activity is reduced See
herbs During storage: GMPs for Fermented Appendix 5
no outgrowth Products
C. Raw meat Yes – No more than 1- For Clostridia, starter Drying is effective for
perfringens and depending log growth C. culture, dextrose, Clostridia after water
C. poultry, on level perfringens; nitrite, and salt activity is reduced See
botulinum spices, no multiplication See page 38 FSIS’ Stabilization
herbs of C. botulinum Guideline
Trichinella Raw pork Yes – Eliminate larvae For Trichinella, For Trichinella, drying
spiralis and (greater zero fermentation can be can be effective in
Toxoplasm infection tolerance effective in combination combination with salt,
a gondii risk: feral with salt and drying smoke, etc. See
(pork) or non- See Porto-Fett (2010) Sausage Treatment
confineme and FSIS Trichinella Methods 1-7 in FSIS
nt swine) Guideline Trichinella Guideline
Molds Any food Maybe No unintentional May apply live mold Drying not effective for
product depending mold growth on culture to prevent molds - rely on
on type finished product growth of undesirable sanitation
molds. Otherwise, rely
on sanitation

15
 Addressing Salmonella, STEC, and Lm: Fermented products have been associated
with Salmonella and STEC outbreaks (see Table 6), and FSIS has detected Salmonella
and Lm in these products (see Appendix 2). The literature shows fermentation and
drying alone do not generally achieve a 5-log reduction of STEC and Salmonella or a 3-
log reduction of Lm (Faith et al., 1997; Faith et al., 1998a; Faith et al., 1998b; Hussein et
al., 2022; Ihnot et al., 1998). However, there are some conditions that have been found
to result in a 5-log reduction as a result of additional interventions such as:

• A high fermentation temperature and a low final pH (Blue Ribbon Task Force, 1996).
• A low pH following fermentation that is maintained or decreases during drying along
with a long drying time (Deibel Laboratories/Chr. Hansen, 2017; Gunvig et al., 2017).
• After fermentation but before drying, apply a low-temperature heat step (Calicioglu et
al., 1997; Hinkens et al., 1996) or FSIS Cooking Guideline for Meat and Poultry
Products (Revised Appendix A) time/temperature/humidity combination.
• Use of high pressure processing (HPP) prior to the fermented sausages reaching a
water activity of less than 0.90 (Balamurugan, 2019).
• Addition of a finishing phase (Hussein et al., 2022) or storage under vacuum at
room-temperature for extended time following fermentation and drying (Calicioglu,
2002; Faith et al., 1997; Faith et al., 1998a; Faith et al., 1998b; Ihnot et al., 1998;
Ingham et al., 2004; Ingham et al., 2005; Porto-Fett et al., 2008).

Critical Steps and Critical Operating Parameters:

Fermentation (see Appendix 6 and Appendix 7 for more information):
• Fermentation temperature, target pH, time to reach target pH, and smoke (if
used).
• Type and use of starter cultures.
• Product characteristics: Casing diameter and shape and product formulation
including salt, sugar (type and level), and use of nitrite or nitrate.

Low-Temperature Heat Step (Optional) (see Appendix 8 for more information)

• Time and temperature: Heating come-up time (CUT), hold time, and temperature
for low temperature heating step.
• Equipment used to generate heat.
• Product characteristics.

Drying (see Appendix 11 for more information):

• Drying room temperature.
• Drying time.
• Target water activity.
• Product characteristics.

Key Points – Fermentation and Drying

Fermentation and drying alone are not particularly effective lethality treatments. When these steps are found
to be effective, there are a lot of critical operational parameters that need to be implemented consistent with
the scientific support. It is not enough to only meet degree-hours, follow a drying or salt-curing method for
Trichinella, and achieve a final water activity for shelf-stability. Degree-hours are intended to control the
outgrowth of S. aureus. To reduce levels of other pathogens such as Salmonella, products often need to be
fermented to a lower pH than 5.3. Also, methods for controlling Trichinella have not been validated for
controlling Salmonella and product may need to be dried longer to reduce levels of Salmonella in the product.
16
Available Scientific Support: Below is a list of available scientific support for the
reduction of Salmonella, STEC, and Lm in fermented products. For detailed summaries
of these common scientific support articles for fermented products see Appendix 12.
Fermented Sausage
• Nickelson, R., II, J. Luchansky, C. Kaspar, and E. Johnson. 1996. Update on dry fermented sausage
Escherichia coli O157:H7 validation research. Blue Ribbon Task Force. Report No. 11-316.
Summer Sausage
• Calicioglu, M., Faith, N.G., Buege, D.R., and Luchansky, J.B. 1997. Viability of Escherichia coli
O157:H7 in Fermented Semi-dry Low-Temperature-Cooked Beef Summer Sausage. J. Food Prot.
60(1): 1158-1162.
Lebanon-Style Bologna
• Getty, K.J.K, Phebus, R.K, Marsden, J.L., Schwenke, J.R., and Kastner, C.L. 1999. Control of
Escherichia coli O157:H7 in Large (115 mm) and Intermediate (90 mm) Diameter Lebanon-style
Bologna. J. of Food Sci. 64(6): 1100-1107.
Salami
• Deibel Laboratories/CHR. Hansen. 2017. Fate of Salmonella Spp., Escherichia coli O157:H7,
Listeria monocytogenes, and Staphylococcus aureus Inoculated Into a Non-Heated and Dried Salami
Product. Available from CHR. Hansen Inc. upon request. https://www.chr-hansen.com/en/contact-us
• Faith, N. G., Parniere, N., Larson, T., Lorang, T.D., Kaspar, C.W., Luchansky, J.B. 1998a. Viability of
Escherichia coli O157:H7 in salami following conditioning of batter, fermentation and drying of sticks,
and storage of slices. J. Food Prot. 61:377-382.
• Hussein, M.H., Burroughs, S., Emch, A.W., Waite-Cusic, J. 2022. Enhancing the Reduction of
Salmonella and Listeria monocytogenes During Traditional Salami Processing by Adding a Finishing
Phase. Food Control. 131: 108432. 2
• Porto-Fett, A.C.S., Call, J.E., Shoyer, B.E., Hill, D.E., Pshebniski, C., Cocoma, G.J., and Luchansky,
J.B. 2010. Evaluation of fermentation, drying, and/or high pressure processing on viability of Listeria
monocytogenes, Escherichia coli O157:H7, Salmonella spp., and Trichinella spiralis in raw pork and
Genoa salami. Int. Journal of Food Micro. 140: 61-75.
Pepperoni
• Hinkens, J.C., Faith, N.G., Lorang, T.D., Bailey, P., Buege, D., Kaspar, C.W., Luchansky, J.B. 1996.
Validation of Pepperoni Processes for Control of Escherichia coli O157:H7. J. Food Prot. 59(12):
1260-1266.
• Ihnot, A.M., Roering, A.M., Wierzba, R.K., Faith, N.G., Luchansky, J.B. 1998. Behavior of Salmonella
typhimurium DT104 during the manufacture and storage of pepperoni. International Journal of Food
Microbiology. 40:117-121.
• Faith, N.G., Parniere, N., Larson, T., Lorang, T.D., Luchansky, J.B. 1997. Viability of Escherichia coli
O157:H7 in pepperoni during the manufacture of sticks and subsequent storage of slices at 21, 4 and
-20°C under air, vacuum and CO2. Int. Journal of Food Micro. 37:47-54.
Soudjouk
• Calicioglu, M., N. G. Faith, D. R. Buege, Luchansky, J.B. 2001. Validation of a Manufacturing Process
for Fermented, Semidry Turkish Soudjouk to Control Escherichia coli O157:H7. J. Food Prot.
64(8):1156-1161.
• Calicioglu, M., N. G. Faith, D. R. Buege, Luchansky, J.B. 2002. Viability of Escherichia coli O157:H7
during manufacturing and storage of fermented, semidry soudjouk-style sausage. J. Food Prot.
65:1541-1544.
• Porto-Fett, A.C.S., Hwang, C.A., Call, J.E., Juneja, V.K., Ingham, S.C., Ingham, B.H., Luchansky, J.B.
2008. Viability of multi-strain mixtures of Listeria monocytogenes, Salmonella typhimurium, or
Escherichia coli O157:H7 inoculated into the batter or onto the surface of a soudjouk-style semi-dry
sausage. Food Microbiology. 25: 793-801.2
Landjäger
• Rivera-Reyes, M., Campbell, J.A., Cutter, C.N. 2017. Pathogen reductions associated with
traditional processing of Landjäger. Food Control. 73: 768-774. 2

2
Studies did not achieve a 5.0-log reduction so either a finishing phase (Hussein et al., 2022) or storage
step under vacuum should be added (see Rivera-Reyes et al., 2017 and Porto-Fett et al. 2008) or
additional supporting documentation (i.e., journal article or challenge study) should be provided.
17
 Overview of Salt-Cured Products

RTE salt-cured meat and poultry products are usually whole muscle products that are
cured with salt and sodium nitrite or nitrate, then air dried, and sometimes smoked (if
desired for certain flavor characteristics). Examples of salt-cured products include
Prosciutto ham, Parma ham, Westphalian ham, Bayonne ham, Serrano ham, Black
Forest ham, country ham, pancetta, coppa, capocolla, bresaola, beef prosciutto,
basturma, duck prosciutto, linguica, and salchichon. Because many products rely on
dry-curing, the guideline focuses on the critical operational parameters of dry-curing.
See examples of scientific support for products in bold in Appendix 13.

Table 4. Overview of hazards of concern establishments should consider in the hazard

analysis during lethality and stabilization and typical controls for salt-cured products.

Hazard Source Adulterant Recommended Salt- Drying

(Yes/No) Target curing/Equalization
Salmonella Raw meat Yes – 5-log reduction Effectiveness for Effectiveness for
STEC (beef) and zero for Salmonella Salmonella, STEC, Lm Salmonella, STEC, Lm
Lm poultry, tolerance and STEC depends on: depends on:
spices, 3-log reduction Curing temperature Drying room
herbs for Lm Curing time temperature
For more Salt coverage of Drying time
information exposed tissue Target water activity
including on Product characteristics Product characteristics
alternative See Appendix 9 See Appendix 11
lethality targets
see Appendix 3
S. aureus Raw meat Yes – No more than 2- For S. aureus, high Drying is effective for
and depending log growth enough brine S. aureus during
poultry on level during concentration/low storage after water
spices, production; enough water activity activity is reduced See
herbs no outgrowth prior to drying Appendix 5
during storage See page 36
C. Raw meat Yes – No more than 1- For Clostridia, Drying is effective for
perfringens and depending log growth C. resting/equalization Clostridia after the
C. poultry, on level perfringens; critical, high enough water activity is
botulinum spices, no multiplication brine concentration/low reduced See FSIS’
herbs of C. botulinum enough water activity Stabilization Guideline
prior to drying, use of
nitrite or nitrate
See page 38
Trichinella Raw pork Yes – Eliminate larvae Effective for Trichinella Effective for Trichinella
spiralis (greater zero with curing, with curing,
and risk of tolerance equalization, and drying equalization, and
Toxoplasm infection per methods for drying per methods for
a gondii for feral or capocollo, hams and capocollo, hams and
(pork) non- pork shoulder picnics, pork shoulder picnics,
confineme boneless pork loins in boneless pork loins in
nt raised and country ham in and country ham in
swine) FSIS Trichinella FSIS Trichinella
Guideline Guideline
Molds Any food Maybe No unintentional Not effective for molds Not effective for molds
product depending mold growth on – rely on sanitation – rely on sanitation
on type finished product

18
Addressing STEC, Salmonella and Lm: Salt-cured products have been associated
with Salmonella outbreaks (see Table 6) and FSIS has detected Salmonella and Lm in
these products (see Appendix 2). There is limited literature available that supports salt-
curing and drying alone achieve a 5-log reduction of STEC and Salmonella or a 3-log
reduction of Lm (Ingham et al., 2006; Genigeorgis and Lindroth, 1984; Reynolds et al.,
2001).

However, there are some examples that have been found to result in a 5-log reduction:
• Dry-curing beef strips, followed by a low-temperature heat step, followed by
drying (Genigeorgis and Lindroth, 1984).
• Dry-curing hams with extended equalization and storage (Reynolds et al., 2001).

Critical Steps and Critical Operating Parameters:

Dry-Curing and Salt-Equalization (see Appendix 9 for more information):
Dry-Curing Salt-Equalization
• Curing temperature. • Equalization temperature.
• Curing time. • Equalization time.
• Salt coverage of exposed muscle • Brine concentration and water
tissue. activity after equalization.
• Product characteristics (e.g., • Product size (diameter or
product size and formulation, thickness).
including salt concentration).

Drying (see Appendix 11 for more information):

• Drying room temperature.
• Drying time.
• Target water activity.
• Product characteristics.
NOTE: High pressure processing (HPP) may also be used for salt-cured hams to add
to the overall process lethality (Perez-Baltar, 2020).
Available Scientific Support: Below is a list of available scientific support for the
reduction of Salmonella, STEC, and Lm in salt-cured products. For detailed summaries
of these common scientific support used for salt-cured products see Appendix 13.
Basturma
• Ingham, S.C., Searls, G., Buege, D.R.. 2006. Inhibition of Salmonella serovars, Escherichia coli
O157:H7, and Listeria monocytogenes during dry-curing and drying of meat: a case study with
basturma. J. Food Safety 26: 160-172.3
• Genigeorgis, C., Lindroth, S. 1984. The Safety of Basturma, An Armenian-type Dried Beef Product
with Regard to Salmonella. Proceedings of the 30th European Meeting of Meat Research Workers.
Bristol, UK. 217-224.
Country Cured Ham
• Reynolds, A.E., Harrison, M.A., Rose-Morrow, R., Lyon, C.E. 2001. Validation of Dry Cured Ham
Process for Control of Pathogens. J. Food Sci. 66:1373-1379.
Bresaola
• Watson, S.C., Gaydos, N.J., Egolf, S.R., Campbell, J.A. 2021. Fate of Escherichia coli O157:H7,
Salmonella spp., and Listeria monocytogenes During Curing and Drying of Beef Bresaola. Meat and
Muscle Biology. 5(1): 14, 1-8.

3
Study did not achieve a 5-log reduction so additional supporting documentation (i.e., additional journal
article or challenge study) should be provided.
19
 Overview of Dried Products

RTE dried meat and poultry products can be comminuted, sliced whole muscle, or
whole muscle products that may or may not be formulated with nitrite, may be smoked,
are usually heated, and are air dried or oven dried. In addition, meat and poultry
products that are freeze dried are also considered by FSIS to be RTE dried products.
Examples of products that are dried as the primary lethality treatment include dried
beef, (some) beef jerky, beef nuggets, steak tenders, kippered beef, meat sticks, turkey
jerky, tasajo, pemmican, pipi kaula, droëwors, biltong, jamon (jambon), longanisa,
(some) saucisson, (some) chorizo, dried soup mixes/soup bases, freeze-dried entrees,
fried pork skins/rinds/cracklings/chicharrones, and lard. See examples of scientific
support for products in bold in Appendix 14.

Table 5. Overview of hazards of concern establishments should consider in the hazard

analysis during lethality and stabilization and typical controls for dried products.

Hazard Source Adulterant Recommended Seasoning/Marination Drying

(Yes/No) Target
Salmonella Raw meat Yes – 5-log reduction Effectiveness for Effectiveness for
STEC (beef) and zero for Salmonella Salmonella, STEC, Lm Salmonella, STEC, Lm
Lm poultry, tolerance and STEC depends on: depends on:
spices, 3-log reduction Product formulation Drying room
herbs for Lm. Antimicrobial temperature
For more application Drying time
information See Appendix 9 Target water activity
including on Product characteristics
alternative See Appendix 11
lethality targets
see Appendix 3
S. aureus Raw meat Yes – No more than 2- Effectiveness of Drying is effective for
and depending log growth seasoning/marination S. aureus during
poultry on level during on S. aureus unclear storage after the water
spices, production; activity is reduced
herbs no outgrowth See Appendix 5
during storage
C. Raw meat Yes – No more than 1- Effectiveness of Drying is effective for
perfringens and depending log growth C. seasoning/marination Clostridia after the
C. poultry, on level perfringens; on Clostridia unclear water activity is
botulinum spices, no multiplication reduced
herbs of C. botulinum See FSIS’
Stabilization Guideline
Trichinella Raw pork Yes – Eliminate larvae Seasoning/marination Drying not effective on
spiralis and (greater zero not effective for its own for Trichinella
Toxoplasm risk of tolerance Trichinella – see FSIS – see FSIS Trichinella
a gondii infection Trichinella Guideline for Guideline for
(pork) for feral or alternatives such as alternatives such as
non- freezing curing or freezing
confineme
nt raised
swine)
Molds Any food Maybe No unintentional Seasoning/marination Drying not effective for
product depending mold growth on not effective for molds – molds – rely on
on type finished product rely on sanitation sanitation
20
Addressing STEC, Salmonella, and Lm: Dried products have been associated with
Salmonella outbreaks outside of the U.S. (see Appendix 14) and FSIS has detected
Salmonella and Lm in these products (see Appendix 2). In general, literature does not
support that drying alone achieves a 5-log reduction of Salmonella and STEC or a 3-log
reduction in Lm. Therefore, additional interventions are often needed to produce a RTE
product. Examples of interventions that may provide additional lethality include
antimicrobial interventions such as lactic or acetic acid (e.g., vinegar) or HPP.
Establishments should be aware that HPP is less effective on intermediate moisture
foods (i.e., those foods that do not require refrigeration to control pathogens)
(Balamurugan, 2019; Perez-Baltar, 2020).

Critical Steps and Critical Operating Parameters:

Marination/Seasoning (see Appendix 10 for more

Key Point – Additional Interventions
information):
• Product formulation. It is not appropriate to add up the
• Antimicrobial application (e.g., concentration, results of two separate studies
pH, coverage, contact time). conducted for the same type of
intervention (such as two acid dips)
Drying (see Appendix 11 for more information):
• Drying room temperature. because the second time the
• Drying time. intervention is used it will likely be less
• Target water activity. effective. This is because any bacteria
• Product characteristics. that survive the first treatment are
likely to be more tolerant to the second
Available Scientific Support: Below is a list of treatment.
available scientific support for the reduction of
Salmonella, STEC, and Lm in dried products. For detailed summaries of these common
scientific support used for dried products see Appendix 14.
Droëwors
• Burnham, G.M., Hanson, D.J., Koshick, C.M., Ingham, S.C. 2008. Death of Salmonella Serovars,
Escherichia coli O157:H7, Staphylococcus aureus, and Listeria monocytogenes During the Drying of
Meat: a Case Study Using Biltong and Droewors. J. Food Safety. 28:198-209. 4

Biltong
• Burnham, G.M., Hanson, D.J., Koshick, C.M., Ingham, S.C. 2008. Death of Salmonella Serovars,
Escherichia coli O157:H7, Staphylococcus aureus, and Listeria monocytogenes During the Drying of
Meat: a Case Study Using Biltong and Droewors. J. Food Safety. 28:198-209. 4
• Karolenko, C.E., Bhusal, Ar., Nelson, J.L., Muriana, P.M. 2020. Processing of Biltong (Dried Beef) to
Achieve USDA-FSIS 5-log Reduction of Salmonella Without a Heat Lethality Step. Microorganisms.
8(5): 791.
• Naidoo, K., Lindsay, D. 2010. Survival of Listeria monocytogenes, and Enterotoxin-Producing
Staphylococcus aureus and Staphylococcus pasteuri, During Two Types of Biltong-manufacturing
Practices. Food Control. 21:1042-1050. 4

4
Studies did not achieve a 5-log reduction or there were methodological issues so additional supporting
documentation (i.e., a journal article or challenge study) should be provided.
21
Post-lethality Considerations

In addition to achieving lethality and shelf-stability using fermentation, salt-curing, and

drying, it is important to ensure contamination and adulteration of products is prevented
after the lethality treatment is complete. Even if the product is RTE shelf-stable,
pathogens may still be able to survive on the product if it becomes contaminated during
handling. RTE foods implicated in foodborne illness are commonly implicated due to
post-processing contamination by bacteria such as S. aureus and Lm either by food
handlers or from the environment.

To ensure that contamination and adulteration of products is prevented after the lethality
treatment, establishments are required to:

• Develop and implement SSOPs (9 CFR 416.11-9 CFR 416.16).

• Maintain sanitation in the RTE area to ensure that food contact surfaces are free
of contamination from Lm and other pathogens, such as Salmonella, in
accordance with 9 CFR part 430.

• Support the safety of non-meat ingredients added part-way through the lethality
treatment (e.g., during lard application during drying of salt-cured hams) or after
the final lethality step is complete (e.g., coating dry or semi-dry sausages in
pepper or rolling sausages in rice flour to give the appearance of an outer
coating of mold). In accordance with 9 CFR 417.2(a)(1) and 417.5(a)(1):

o Establishments must determine what potential hazards are associated

with the non-meat ingredients at the step in the process when the
ingredients are “received” into the food safety system.

o Establishments must also document any controls needed to support

decisions about those hazards such as Letters of Guarantee (LOGs),
Certificates of Analysis (COAs), or other information (e.g., sampling by
the receiving establishment).

Because fermented, salt-cured, and dried products rely on multiple hurdles,

establishments are required to identify the step in the process when lethality is complete
according to their scientific support for purposes of complying with the Listeria Rule (9
CFR 430) which addresses control of Lm in the post-lethality environment. Identifying
the step where lethality is achieved will ensure that:

• the establishment identifies all possible surfaces that contact the food after the
post-lethality step for sampling (required under 9 CFR 430.4(b)(2)(iii)(A) and
(3)(i)(A)); and

• any post-lethality treatments are designed and implemented appropriately.

Identifying when lethality ends and the post-lethality environment begins is particularly
important since some treatments, such as drying, extended storage, or HPP, can be
used as part of the multi-hurdle lethality or as a post-lethality treatment or both. For
22
example, if a study shows a 5-log reduction in STEC, Salmonella, and Lm occurs after
18 days of drying, then the establishment would identify the lethality treatment ends
after 18 days of drying and the post-lethality environment begins when the product is in
the drying room on day 19. The establishment would also identify any food contact
surfaces that the product contacts after 18 days of drying (e.g., racks, aprons,
packaging machines, carts) in its Lm Control Program to address potential post-lethality
Lm contamination. This would include any food contact surfaces the product contacts
during drying that are also in contact after the 18 days of drying are complete, such as
racks. The establishment may also choose to implement a validated post-lethality
treatment after 18 days of drying, such as additional 60 days of storage under vacuum
at 70°F to meet the requirements of Alternative 2, Choice 1 (Alt. 2a) [or Alternative 1 if
the establishment can also support the product’s water activity is below the growth limit
of Lm]. In this example, drying would be part of the lethality treatment and extended
storage would be a post-lethality treatment. Examples of post-lethality treatments that
have been validated for fermented products include storage under vacuum at
refrigerated temperatures (Faith et al., 1997; Faith et al., 1998a; Faith et al., 1998b;
Ihnot et al., 1998; Ingham et al., 2004) and pasteurization by submersion heating
(Roering et al., 1998). HPP has been validated as a post-lethality treatment for
products such as salt-cured ham (Perez-Baltar et al., 2020).

Further guidance on post-processing handling and sanitation for RTE products is

available in the FSIS Cooking Guideline for Meat and Poultry Products (Revised
Appendix A) and the Guidelines to Control Listeria monocytogenes in Post-Lethality
Exposed Ready-to-Eat Meat and Poultry Products. The Lm guideline also contains
more information on post-lethality treatments and when fermentation and drying may be
considered as antimicrobial processes.

23
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Appendix 1: Labeling Considerations for Not Ready-to-Eat (NRTE)
fermented, salt-cured, and dried products.

The following are additional labeling considerations for NRTE fermented, salt-cured,
and dried meat and poultry products.

Many of the products covered in this guideline can be classified by establishments as

RTE or NRTE and as shelf-stable or not (i.e., it needs to be refrigerated or frozen by the
consumer throughout storage or once the package is opened).
as described below:

• Fermented
o Dry and semi-dry fermented sausages – intended use may be RTE or NRTE
except for pepperoni and salami which typically have an intended use of RTE.

• Salt-cured
o Basturma – intended use may be RTE or NRTE.
o Country cured ham – intended use may be RTE or NRTE.
o Bresaola – intended use is typically RTE.

• Dried
o Biltong – intended use is typically RTE.
o Droëwors – intended use is typically RTE.

For those products where the intended use may be NRTE, the product must bear safe
handling instructions (SHI) per 9 CFR 317.2(k)(1) or 9 CFR 381.125. Additional labeling
features are required for those products described in this guideline such as dry and
semi-dry fermented sausages, basturma, and country cured ham that may have
the appearance of an RTE product (e.g., as a result of the fermentation, salt-curing, or
drying steps) but that are classified as NRTE by the establishment. Because these
products require cooking by the consumer for safety, FSIS requires a prominent
statement on the principal display panel, which may include statements such as,
“Uncooked, Ready to cook, Cook before eating, Cook and serve” or “Needs to be fully
cooked.”

To help ensure that NRTE products that appear RTE are cooked, FSIS also
recommends that validated cooking instructions be provided on the label. The
Guidelines to Control Listeria monocytogenes in Post-Lethality Exposed Ready-to-Eat
Meat and Poultry Products contains further information on other labeling features that
should be included on NRTE products that appear RTE. Establishments should be
aware that many fermented, salt-cured, and dried products have product characteristics
such as intermediate and low water activity that result in the typical consumer cooking
instructions that are provided on the label being insufficient if the product is NRTE. For
example, FSIS recommends that consumers cook raw beef sausages to 160°F. This
recommendation is based on research demonstrating that achieving this temperature
would result in at least a 6.5-log reduction in Salmonella as shown in FSIS Cooking
Guideline for Meat and Poultry Products (Revised Appendix A) (Blankenship, 1978;
Goodfellow and Brown, 1978).

33
NOTE: It would not be appropriate to recommend a NRTE salt-cured and dried product
such as basturma be cooked to 160°F using FSIS Cooking Guideline for Meat and
Poultry Products (Revised Appendix A) unless the intended use is to be cooked under
moist conditions to rehydrate the surface.

Once a product is salt-cured or dried, any Salmonella remaining in the product would
have increased thermal tolerance from surviving the drying process and additional heat
would be needed to destroy remaining bacteria. Establishments that use consumer
cooking in conjunction with its purchasing specifications and its own preventive
measures employed during further processing to support decisions related to hazards in
NRTE fermented, salt-cured, and dried products, need to have on-file documentation
supporting their decisions (9 CFR 417.5(a)(1)). Such documentation may include
documents describing the customary preparation practices for the safe consumption of
the product and the basis for the establishment's determination that these practices
constitute customary preparation. Such support could include a challenge study
validating the cooking method recommended results in a safe product (e.g., a 5-log
reduction in Salmonella) or a science-based justification for why the cooking method
recommended results in a safe product (e.g., instructions that include a step where the
product is immersed in water such as those typically used for country cured hams would
rehydrate the product). For more information on conducting challenge studies see
Appendix 15: Designing Challenge Studies for Fermented, Salt-Cured, and Dried
Products.

If an establishment identifies the intended use as NRTE for products such as pepperoni,
salami, bresaola, biltong, and droëwors where the intended use is typically RTE, the
establishment must have on-file documentation supporting their decisions (9 CFR
417.5(a)(1)). This support must address how the establishment can ensure the
consumer will properly cook the product (9 CFR 417.5(a)(1)), particularly if there is
evidence such as marketing materials or recipes commonly indicating the product is
RTE.

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 Appendix 2: Biological Hazards of Public Health Concern for RTE Shelf-
Stable Fermented, Salt-Cured, and Dried Products

The following are further considerations regarding hazards of public health concern for
RTE shelf-stable fermented, salt-cured, and dried products that can help inform an
establishment’s hazard analysis decision-making.

Salmonella, STEC, and Lm

Meat and poultry products may become contaminated with Salmonella, STEC (in beef),
and Lm, from cross-contamination during the slaughter/dressing process or in the
processing environment when insanitary conditions are present. In addition, spices and
herbs can also be contaminated with Salmonella, STEC, and Lm. These pathogens can
survive typical fermentation, salt-curing and drying treatments. FSIS has detected
Salmonella and Lm in fermented and dried products.

• Between 2010-2018, the average percent positive for Salmonella in RTE

fermented or dried products was 0.09% (12/12,684) compared to 0.05% for other
RTE products (47/99,038).
• Between 2010-2018, the average percent positive for Lm in RTE fermented or
dried products was 0.23% (31/13,609) compared to 0.32% for other RTE
products (333/106,827).

A few outbreaks associated with Salmonella and E. coli O157:H7 have occurred in the
U.S. that were associated with fermented and salt-cured meats as shown in Table 6.
Table 6. Salmonella and E. coli O157:H7 Foodborne Illness Outbreak History in Fermented
and Salt-Cured, and Dried Ready-to-Eat Meat Products Produced in the U.S.

Products Year Disease- Case-Patients; Recall Process Suspected

Affected Causing States (Yes/No) Type Root Cause
Organism
Salami 2021 Salmonella I 34; 10 states Yes Fermented Under-
sticks 4,[5],12:i:- and dried processing
Italian-style 2021 Salmonella 40; 17 states Yes Fermented Under-
Meats Infantis and and dried processing
Tyhphimuriu
m
Lebanon 2011 E. coli 14; Yes Fermented Under-
Bologna O157:H7 5 states processing
Prosciutto, 2010 Salmonella 272; Yes Salt-cured Contaminated
Capocollo, Montevideo 45 states and red and black
calabrese, Fermented pepper
sopressata and dried
Lebanon 1995 Salmonella 26; Yes Fermented Under-
Bologna Typhimurium 1 state processing
Salami 1994 E. coli 23; Yes Fermented Under-
O157:H7 2 states and dried processing
Basturma 1982 Salmonella Unknown; Unknown Salt-cured Unknown
1 state and dried

35
Several outbreaks have also been associated with fermented, salt-cured, and dried
meat products outside of the U.S., with five outbreaks occurring in Europe between
2018 and 2020:

• In November 2020, a Salmonella outbreak in France was linked to a dry sausage

produced in France (York, 2020).
• In September 2020, a Salmonella outbreak in France was linked to fuet sausage
produced in Spain (Whitworth, 2020).
• In July 2019, a Salmonella outbreak in France was linked to coppa produced in
Italy.
• In November 2018, a Shiga toxin-producing Escherichia coli (STEC) outbreak
(O26:H11) in Denmark was attributed to a beef salami product (Whitworth, 2019).
• In October 2018, an outbreak of Salmonella Typhimurium in Denmark was
potentially attributed to a spiced pork sausage.

Other outbreaks of note outside the U.S. include a 2015 Salmonella outbreak in
Australia associated with duck prosciutto (Draper et. al, 2017) as well as a Salmonella
outbreak associated with biltong consumption in London in 2008 (Mindlin, 2013).

The presence of Salmonella, Lm, or E. coli O157:H7 in fermented and dried meat
products may indicate under processing/insufficient lethality due to lack of a
cooking step (Farber et al., 1988). Although Lm contamination typically indicates post-
lethality contamination, its presence in fermented, salt-cured, and dried meat and
poultry products may also indicate under processing/insufficient lethality. This is
because Lm can be present on raw meat and poultry products 5 and other ingredients
and survive fermentation and drying because Lm is very tolerant to those types of
lethality treatments. For more information on Lm prevalence in raw meat and poultry
products, see FSIS’ Baseline Data Reports.
NOTE: FSIS tested dry and semi-dry fermented sausages for E. coli O157:H7
prevalence between 1994 and 2011. During this time, FSIS did not have a positive E.
coli O157:H7 test result from over 10,000 samples of these products. Therefore, FSIS
discontinued testing. While FSIS is aware there can be problems associated with STEC
in these products, such as the March 2011 Lebanon bologna E. coli O157:H7 outbreak,
the Agency determined samples are not collected at a frequency high enough to detect
such processing issues that tend to occur intermittently and at a low frequency. These
problems are most often found during FSIS food safety assessments (FSAs) or during
other in-depth establishment reviews.

Staphylococcus aureus

Staphylococcus aureus (S. aureus) can contaminate raw meat and poultry from the
animal hide, skin, or tissue during slaughter. After slaughter and after cooking, RTE
meat or poultry products can be contaminated from handling by individuals carrying the

5
FSIS baseline testing has identified Lm on all raw meat and poultry source materials at levels ranging
from 4.1% on steer and heifer carcasses to 7.4% on market hogs and as high as 41.1% in raw ground
chicken.
36
organism. S. aureus is of concern during storage of shelf-stable products because it is
salt-tolerant and can grow at a lower water activity than other bacterial pathogens.

S. aureus is a concern during fermentation, salt-curing, and drying because the addition
of the ingredients (salt, sodium nitrite, sodium nitrate) creates microbial inversion
when the environment favors Gram-positive bacteria such as S. aureus as opposed to
Gram-negative bacteria such as STEC and Salmonella (which grow better on raw
product without ingredients added).

• For fermented products, the degree-hours concept is typically used to ensure

that growth of S. aureus is limited during fermentation when the temperature is
over 60 °F (15.6°C) (the critical temperature at which staphylococcal growth
begins).

o Degrees are measured as the excess over 60°F (15.6°C). Degree-hours

are the product of time at a particular temperature and the “degrees.”
Degree-hours are calculated for each temperature used during
fermentation. The limitation of the number of degree-hours depends upon
the highest temperature in the fermentation
process prior to the time that a pH of 5.3 or less
is attained.

o Good Manufacturing Practices for Fermented

Dry and Semi-dry Sausage Products describes
the degree-hours concept in detail.
KEY DEFINITIONS
o The degree-hour guidance does not identify the
maximum amount of growth of S. aureus
expected when degree-hours are met; however, Degree-hours is the amount
FSIS considers meeting degree-hours to limit of time in hours above 60°F
growth to safe levels (i.e., 2-logs or less) (Smith (the critical temperature at
and Palumbo, 1978). which staphylococcal growth
effectively begins) an
• For salt-cured products, the salt-cure is allowed to establishment’s fermentation
equilibrate so that it penetrates throughout the raw process can take at a
source materials so that S. aureus growth is prevented specific temperature to
during drying when temperatures are elevated. reduce the pH to 5.3 or
below in order to control S.
o Generally, a 10% brine concentration aureus growth.
following equalization will prohibit S. aureus
enterotoxin production when the temperature is increased during drying
(Reiman et al., 1972).

o S. aureus may also contaminate products post-lethality during handling

and is also of concern during storage of shelf-stable products because it
can grow at a lower water activity than other bacterial pathogens.

Between 1994 and December 2002, FSIS tested 3,105 RTE products for the presence
of staphylococcal enterotoxins (< 1 nanogram (ng) staphylococcal enterotoxins toxin per

37
gram or ml of sample). FSIS stopped testing RTE products for staphylococcal
enterotoxins in January 2003 because FSIS did not find any positive samples. These
negative findings are likely due in large part to the widespread use of commercial starter
cultures and addition of fermentable sugars as well as education of producers by starter
culture suppliers and trade associations in best practices for production of fermented
meats (Smith and Palumbo, 1983), practices developed in response to outbreaks in the
early 1970’s (CDC, 1971a; CDC 1971b; CDC, 1975). For this reason, it is important that
establishments continue to ensure processes are sufficient to limit S. aureus growth and
enterotoxin production (e.g., by following Good Manufacturing Practices for Fermented
Dry and Semi-dry Sausage Products). For more information on FSIS’ method for testing
RTE products for the presence of staphylococcal enterotoxins see the Microbiology
Laboratory Guidebook Method 39.03 available at: https://www.fsis.usda.gov/news-
events/publications/microbiology-laboratory-guidebook.

Clostridium perfringens and Clostridium botulinum

Meat and poultry products may become contaminated with C. perfringens and C.
botulinum during the slaughter and dressing process as a result of cross-contamination
in the processing environment when insanitary conditions are present. In addition,
spices and herbs can contribute to the spore counts in raw formulated heat-treated
meat and poultry products. For example, in one survey, C. perfringens spores were
isolated from 43 of 54 (80%) different spices and herbs (Juneja and Sofos, 2010).

There have been several outbreaks in Europe associated with non-proteolytic C.

botulinum and home-prepared (salted) ham (Mazuet et al., 2015; Peck et al., 2015).
However, under commercial processing conditions, spores of Clostridia will generally
not germinate and grow out during production of dried meats because the
microbiological environment of such products does not allow for the outgrowth of
spores. Specifically, the acidity (reduced pH) and dryness (reduced water activity) of
dried meats create conditions that prevent germination. FSIS’ Stabilization Guideline
for Meat and Poultry Products (Revised Appendix B) contains recommended pH and
water activity values that will prevent the growth of C. perfringens and C. botulinum.
Other controls include salt concentration, presence of lactic acid bacteria, and use of
nitrate and nitrite. Use of sodium nitrite (50 parts per million (ppm)) or water activity of
0.92 or below are sufficient to ensure C. botulinum will not grow in these fermented,
salt-cured, and dried products (Reynolds et al., 2001; Johnston et al., 1969; and
Tompkin, 1976). The resting or equalization phase is a critical step for inhibiting C.
botulinum growth in dry-cured hams (Merialdi et al., 2016). In fermented products such
as summer sausage, the use of a starter culture, dextrose, and nitrite were needed to
inhibit C. botulinum growth (Christiansen et al., 1975). The addition of at least 100 ppm
nitrite/nitrate and at least 2.5% salt can also be used to control C. botulinum growth in
fermented products during the fermentation and drying steps (CFIA, 2020).

Trichinella and Toxoplasma gondii

Trichinella spiralis (T. spiralis) is a parasite that infects both humans and animals.
Establishments producing RTE fermented, salt-cured, and dried pork products should
be aware this is a hazard to consider as part of the hazard analysis, particularly if using
pork from source materials derived from feral or non-confinement raised pigs. This
guideline does not go into detail regarding how to address T. spiralis (or other potential
38
parasitic hazards such as Toxoplasma gondii) because control measures are addressed
in the FSIS Guideline for the Prevention and Control of Trichinella and Other Parasitic
Hazards in Pork and Products Containing Pork. FSIS’ Trichinella Guideline contains one
reference (Gamble and Hill, 2012) that addresses the effectiveness of curing for the
destruction of T. gondii in pork products. T. gondii in dry-cured pork sausage may also
be addressed with other scientific support (Hill et al., 2018). Because there are no
published studies comparing the lethality rate of Salmonella to the destruction of
Trichinella in dried, salt-cured, or fermented products, it is not appropriate to apply
scientific support conducted with Salmonella to support decisions related to Trichinella
and it is also not appropriate to apply scientific support conducted with Trichinella to
support decisions related to Salmonella. Relying on a minimum number of drying days
for Trichinella elimination without validating the number of drying days needed for
Salmonella reduction was a contributing factor in two Salmonella outbreaks associated
with Italian-style fermented meats (FSIS, 2022).

What About Mold?

Molds may grow on products like country cured ham (also known as country ham)
during the curing process and drying process because the associated high salt, low
temperatures, and environmental conditions do not inhibit these organisms. Most molds
found on dry-cured products are harmless, although some molds are undesirable, and
may cause allergic reactions and respiratory problems. A few types of molds, under the
right conditions, produce mycotoxins, which are poisonous substances that can make
consumers sick. Undesirable molds may also affect product quality as they can
breakdown proteins and fats. Adherence to SSOPs is critical to prevent undesirable
mold growth during processing and to reduce airborne mold spores in fermentation and
aging rooms (Quintavalla, 2010). During storage, measures to prevent undesirable mold
growth may include using short inventory pull dates, low pH, sufficiently low water
activity, antimycotics, coatings, packaging, or any combination of these measures.

If mold is present during processing, depending on the type of mold, establishments

may package the product with the mold on or scrub off the mold with a stiff vegetable
brush to maintain wholesomeness. FSIS does not recommend washing mold off using
hoses that can result in cross-contamination from the environment to the product. Dry-
white mold, often seen on fermented and dried sausages, is generally considered to be
a good mold because it can prevent “bad mold” from growing although color is not
necessarily an indicator of “bad mold”. Sometimes a commercial mold culture will be
added prior to fermentation to actively apply a live mold culture to prevent the growth of
undesirable molds and for flavor development. For more information on the impact of
applying a commercial mold culture, see page 71. If a mold culture is not applied, and
mold growth is desirable for quality reasons, establishments must determine the
hazards reasonably likely to occur (9 CFR 417.2) and support the decision-made (9
CFR 417.5). As part of that decision-making the establishment must consider the effect
mold plays in the process and needs to maintain documentation supporting that they
are controlling or preventing any food safety hazards that might occur during this
process.

39
Appendix 3: Lethality and Shelf-Stability Targets

Lethality Targets (Salmonella, STEC, and Lm)

The lethality treatment (i.e., the combination of hurdles or steps) for RTE shelf-stable
meat and poultry products should achieve at least a 5.0-log reduction of Salmonella and
at least a 5.0-log reduction for STEC including E. coli O157:H7 for products containing
beef as recommended in the FSIS Cooking Guideline for Meat and Poultry Products
(Revised Appendix A). In addition to Salmonella, the lethality treatment of RTE shelf-
stable meat and poultry products should achieve at least a 3.0-log reduction in Lm,
although a 5.0-log reduction or greater is desirable for providing an even greater safety
margin for ensuring that Lm does not grow to detectable levels during storage.
However, establishments are not required to validate their process achieves reductions
in Lm (or STEC for products containing beef) if it achieves sufficient reductions in
Salmonella as indicated in the FSIS HACCP Systems Validation Guideline.

Unlike cooking; however, research has shown that:

• STEC including E. coli O157:H7 and Lm are more tolerant than Salmonella
during the fermentation and drying steps of dry/semi-dry fermented sausages
(Hussein, et al., 2022; Ihnot et al., 1998; Porto-Fett et al., 2010; McKinney,
2019).

• Lm is more tolerant than Salmonella during the drying step of dried and salt-
cured meat and poultry products (Porto-Fett et al., 2010; Reynolds et al., 2001).

Therefore:

• If an establishment’s scientific support is only based on reductions in Salmonella

and the establishment has a STEC or Lm product positive either through its own
testing or FSIS’ testing or is associated with an outbreak of these pathogens, the
Agency would require the establishment as part of its corrective actions to
validate for the other pathogens unless it can support the cause of the positive
was post-lethality contamination.

• FSIS also recommends establishments conducting new challenge studies

(studies where the product is inoculated with bacteria to determine the processes
effectiveness) determine the log reductions in Salmonella, STEC, (in products
containing beef) and Lm. If an establishment is only able to include one
pathogen, it may include either Salmonella, STEC, or Lm although FSIS
recommends choosing Lm or STEC (for products containing beef) due to their
increased survival in response to fermentation and drying.

• Because STEC have been shown to be more tolerant to fermentation and drying
than Salmonella and have been shown to have similar tolerance to Lm, FSIS
would not object to establishments using research that only demonstrates E. coli
O157:H7 reductions in beef and does not include Salmonella or Lm.

40
 • If an establishment’s scientific support includes Lm and STEC, the support
should demonstrate at least a 3.0-log reduction in Lm and at least a 5-log
reduction in STEC to be considered adequate.

• If an establishment’s scientific support only includes Lm (and not Salmonella or

STEC), FSIS recommends the support should demonstrate at least a 5.0-log
reduction in Lm in order to support at least a 5.0-log reduction in Salmonella and
STEC (in beef) is achieved.

Establishments can demonstrate a 5.0-log reduction in Salmonella (and STEC in

products containing beef) by using a combination of hurdles. However, if one of those
hurdles that is used as part of the overall lethality is only applied to the meat or poultry
raw materials (such as an antimicrobial intervention that is only applied to the raw meat
or poultry component), establishments should provide support that the pathogen load on
any non-meat ingredients has also been reduced or that those ingredients are free of
pathogens (e.g., by using COAs that report lot by lot test results).

What Options do I have if my Steps/Hurdles don’t

Achieve a 5.0-log Reduction?

Establishments that are unable to demonstrate a 5-log reduction

in Salmonella or STEC (in products containing beef) also have
the option of applying alternative lethalities. One accepted
alternative lethality is based on “Option #5” from The Blue
KEY DEFINITIONS
Ribbon Task Force in which the raw batter of sausage is tested
in conjunction with the application of a process that achieves at
least a 2.0-log reduction in the hazard of concern. This option Alternative lethality is a
was developed for comminuted products where the entire raw lethality target or log
batter with all the raw ingredients (e.g., spices, flavor, salt, reduction that is different
sugar, nitrite) are tested together. The raw batter testing option from FSIS recommendations,
is discussed in detail in The Blue Ribbon Task Force document. but achieves an equivalent
probability that no viable
Importantly, when using this option, each and every lot of raw
Salmonella organisms or
batter should be tested following the recommendations for
sample size and number of samples collected per lot in The other pathogens of concern
Blue Ribbon Task Force document. This option provides less remain in the finished
assurance of product safety, so it is important that the raw product when properly
material testing provides a high degree of confidence that there implemented as described in
are no hazards of concern present. Although testing every lot of the supporting scientific
raw batter may not be practical for every small and very small
documentation.
establishment, it was recommended originally by the Blue
Ribbon Task force as “the most practical solution to assuring the
safety of certain dry fermented sausage products” and
represents the best alternative when a 5.0-log reduction cannot
be supported.

FSIS has received a number of askFSIS questions regarding

“Option #5” from The Blue Ribbon Task Force and based on
41
these has the following guidance related to applying the option to dry/semi-dry
fermented sausage products containing beef:

• Establishments can test the raw batter of beef products for STEC including E.
coli O157:H7 only as described in the Blue Ribbon Task Force document.

• Because STEC are more tolerant than Salmonella during the fermentation and
drying steps of dry/semi-dry fermented sausages as previously described,
establishments producing dry/semi-dry fermented sausage products do not have
to also test the raw batter for Salmonella or validate the process achieves a
specific log reduction of Salmonella.

• FSIS also would not object to establishments testing the batter of beef containing
products for Salmonella and support that the process achieves at least a 2.0-log
reduction in Salmonella given Salmonella has been used as an indicator. If an
establishment’s scientific support is only based on reductions in Salmonella and
the establishment has a STEC or Lm positive either through its own testing or
FSIS’ testing or is associated with an outbreak of these pathogens, the Agency
would require the establishment as part of its corrective actions to validate for the
other pathogens unless it can support that the cause of the positive was post-
lethality contamination.

FSIS has the following guidance for products not containing beef (e.g., those containing
pork) that are unable to demonstrate a 5.0-log reduction in Salmonella or Lm:

• Establishments can test the raw batter for Salmonella and support the process
achieves at least a 2.0-log reduction in Salmonella without demonstrating that
specific reductions in Lm are achieved.
o Because Salmonella is less tolerant than Lm during the fermentation and
drying steps of dry/semi-dry fermented sausages, if an establishment’s
scientific support is only based on reductions in Salmonella and the
establishment has a Lm product positive either through its own testing or
FSIS’ testing or is associated with an outbreak, the Agency would require
the establishment as part of its corrective actions to validate for Lm unless
it can support that the cause of the positive was post-lethality
contamination.

• Establishments can test the raw batter for Lm and support the process achieves
at least a 2.0-log reduction in Lm without demonstrating that specific reductions
in Salmonella are achieved.
o Because Lm are more tolerant than Salmonella during the fermentation
and drying steps of dry/semi-dry fermented sausages as previously
described, establishments producing dry/semi-dry fermented sausage
products do not have to also test the raw batter for Salmonella or validate
the process achieves a specific log reduction of Salmonella.

In terms of the specific testing, the option in The Blue Ribbon Task Force document
recommends fifteen 25-gram (g) samples be taken from across the lot (regardless of lot
size or weight) and indicates further research is needed to establish the limits of

42
compositing. For Salmonella and E. coli O157:H7, FSIS recommends compositing up
to three – 25-g samples (total 75 grams) for a total of 5 analyses, although
establishments may also be able to support compositing all fifteen – 25-g samples (total
375 grams). If establishments choose to test for Lm, FSIS recommends compositing up
to 5 samples (total 125g) for a total of 3 analyses. When compositing, establishments
should ensure the method has been validated for the larger test portion.

How Can I Apply the Concept of Raw Batter Testing to Whole Muscle Products?

FSIS has identified that establishments producing products using whole muscle cuts
(e.g., biltong) are also unable to achieve at least a 5-log reduction of hazards of
concern. FSIS recommends as an alternative, that every lot of the raw materials are
tested in conjunction with a process validated to achieve at least a 2.0-log reduction.
For whole muscle cuts, establishments should test both the meat or poultry and non-
meat ingredients to support that the incoming pathogen levels are low and that a 2.0-log
reduction would be sufficient to achieve product safety with confidence. For the non-
meat ingredients, establishments may be able to support the incoming pathogen load of
the raw materials is low through means other than their own testing. For example, when
using spices, an establishment may receive a COA or LOG that indicates how each lot
of ingredients is processed, tested, or otherwise treated to ensure its safety.

For the meat or poultry component, FSIS recommends establishments collect fifteen –
25-g samples of the meat or poultry whole muscle cut using excision sampling. This
sample size is the same as that recommended for the raw batter testing option and the
International Commission on Microbiological Specifications for Foods (ICMSF) case 13
sampling plan (for more information on the ICMSF sampling plans, see page 120 of the
FSIS Guideline: Controlling Listeria monocytogenes in Post-Lethality Exposed RTE
Meat and Poultry Products). FSIS recommends that excision samples are used for
intact whole muscle cuts, but not for injected, vacuum-tumbled, or mechanically
tenderized (by needle or blade) products. If the raw meat or poultry component is
processed in such a way that it is made non-intact, the sampling should include
excisions of both interior and exterior portions. The reason for this is because research
has shown that such processing techniques push bacteria into meat and are less likely
to be detected on the surface. One way to excise the surface and interior of meat is with
a sanitized coring device (Ulbrich, 2015; Luchansky, 2008). For this sampling technique,
establishments should use a sterile coring device with a 10 cm2 diameter (surface area)
to obtain core samples. Sanitized knife and forceps are recommended to cut segments
of the exterior and interior portions of the core sample. For further guidance on this
method see Luchansky et al., 2008. Guidance regarding selection of organisms for
testing and validation as well as for compositing of raw batter samples provided for raw
batter testing provided earlier in this document on page 42 also applies to sampling and
testing of whole muscle raw materials.

43
Shelf-Stability Targets

Fermented, salt-cured, and dried products typically achieve

shelf-stability through a combination of hurdles such as
reduced pH, reduced water activity, or a combination of both,
along with other extrinsic factors, such as reduced oxygen KEY DEFINITIONS
and packaging. S. aureus is the main pathogen of concern
during storage of shelf-stable products because it can grow at
Moisture Protein Ratio
a lower water activity than other pathogens. To achieve of
(MPR) expresses the percent
shelf-stability, no outgrowth of S. aureus may occur in the
moisture divided by the
product. Minimizing available water (e.g., by achieving a
percent protein. MPR is
sufficiently low water activity) is necessary to achieve shelf
commonly used in the U.S. to
stability, provided measures are taken to address undesirable
classify dried sausages and
mold growth. Such measures to prevent undesirable mold
other meat products.
growth may include using short inventory pull dates, low pH,
Although MPR values
antimycotics (such as potassium sorbate sprays and dips),
indicate the degree of
coatings, packaging, or any combination of these measures.
product drying, they are not
necessarily indicative of
For examples of scientific support that can be used to support
microbial safety or product
shelf-stability see Appendix 5.
shelf-stability because they
do not take into account
NOTE: FSIS has a regulatory definition of shelf-stability in (9
availability of the water.
CFR 431.1) but this only applies to thermally processed,
commercially sterile products and does not apply to the types
of products described in this guideline.

Can I Use Finished Product Testing Alone if I Don’t have

Scientific Support for my Process?

No. FSIS does not consider test and hold (also sometimes
described as “Option #3” from The Blue Ribbon Task Force
document) as acceptable support because it relies on finished
product testing alone and does not support a specific log
reduction in levels of target pathogens. So, relying on finished
product testing alone is not consistent with the concepts of
HACCP in which hazards are to be prevented or controlled by
the HACCP system. In addition, water activity testing alone
would also not be an appropriate way to support the safety of
RTE meat and poultry products for the same reasons.

NOTE: An establishment may rely on finished product testing

alone to support product safety for a limited time during the
initial validation period (90 calendar days) if it is in the process
of identifying alternative scientific support (i.e., journal articles
or challenge studies).

44
Appendix 4: Considerations for Different Types of Scientific Support

The following addresses considerations for different types of scientific support that may
be used to support the lethality or shelf-stability of fermented, salt-cured, or dried meat
and poultry products.

Peer-Reviewed Scientific or Technical Data or Information

Peer-reviewed journal articles, many of which are discussed throughout this document,
are available to support the lethality and shelf-stability of fermented, salt-cured, and
dried RTE meat and poultry products. When establishments are reviewing the literature,
they should thoroughly evaluate whether the critical operational parameters used in the
support match those used in the actual process. FSIS recommends that establishments
use a single supporting document to support the effectiveness of its lethality steps. If
multiple studies are used together to support the same process, the establishment will
need to support that the new combination of parameters would be as effective as those
studied in the individual articles or documents (80 FR 27557).

Pathogen Modeling Programs

Establishments should not rely on the results of pathogen modeling programs alone
unless those models have been validated. The following are validated models FSIS
recommends for supporting decisions related to fermented, salt-cured, or dried meat
and poultry products:

• University of Wisconsin Shelf-Stability Predictor available at:

https://meathaccp.wisc.edu/ST_calc.html.
o This model has been validated for estimating the likelihood of S. aureus
and Lm growth.
o Input parameters are pH and water activity.
o For more information see Appendix 5.

• Danish Technological Institute (DMRI) ConFerm model available at

http://dmripredict.dk (Gunvig et al., 2016)
o This model has been validated for estimating the reductions of
Salmonella, STEC, and Lm in fermented products.
o At most, the model can be used to support a 3.0-log reduction of
Salmonella, STEC, and Lm.
o Should only be used for products with matching critical operational
parameters as described in Gunvig et al., 2016.
o Input parameters are: fermentation temperature, % salt in the formulation,
ingoing nitrite ppm, pH at the start before fermentation, pH at 48 hours,
final pH, % weight loss in product in the beginning and after drying, total
process time (fermentation + drying), and water % in final product (as
determined through laboratory analysis).

45
 • DMRI Staphtox Predictor (Version 1.0) available at: http://dmripredict.dk/
o This model has been validated (Gunvig et al., 2017) to predict the growth
of S. aureus and potential toxin formation during mild heat treatment and
during constant temperature fermentation of sausages (aw>0.96).
o Input parameters are NaCl in product, KCl in product, sodium nitrite in the
recipe/ingoing, % water in product (as determined through laboratory
analysis), time, pH, and temperature.
o For more information on using this model to evaluate to evaluate product
safety during fermentation deviations, see Appendix 7.

Other models are available for estimating reductions in bacterial pathogens of concern
in fermented, salt-cured, or dried meat and poultry products, such as the E. coli
O157:H7 survival models for fermented sausage that are part of the Agricultural
Research Service (ARS) Pathogen Modeling Program (PMP) Online and the Meat and
Livestock Model (MLA) E. coli inactivation model in Fermented Meat. However, these
other programs are not considered validated at this time because the results of the
modeling programs have not been compared to the results of actual studies to
determine their accuracy. These models can be useful for providing an initial estimate of
reductions in levels of pathogens prior to conducting a challenge study. Establishments
should not rely on these models alone as scientific support unless they have additional
support.

Challenge or Inoculated Pack Studies

When establishments want to use unique processes that are not supported by the
literature, because there is no literature available or the process used is significantly
different from that studied, a challenge study may be needed to demonstrate the safety
of the process. General guidance on conducting challenge studies can be found on
page 8 of the FSIS HACCP Systems Validation Guideline. Specific considerations for
the design of challenge studies for fermented, salt-cured, and dried products are
included in Appendix 15.

Data Gathered by the Establishment In-Plant

When an establishment has a scientific support document such as a peer-reviewed

journal and it wants to use different critical operational parameters, it may consider
collecting in-plant microbiological data to support the new combination of steps to
achieve adequate reduction in bacterial pathogens of concern, particularly if data or
scientific principles are not available to support the changes. For example, if an
establishment producing a dried meat product identifies a journal article that matches its
process, but it intends to use a slightly lower drying temperature (e.g., 2 or 3°F lower
than that used in the support), it may choose to collect in-plant microbiological data to
provide additional support for its process. In this case, the establishment would take a
statistically based number of samples each day it produces the product during a 90-
calendar day period and analyze the finished product for Salmonella, STEC (including
E. coli O157:H7 in products containing beef), and Lm. If the establishment produces the
product less than 13 days within a 90-calendar day period, it should continue to test
product until 13 different lots have been tested and found negative for Salmonella,
STEC (including E. coli O157:H7 in products containing beef), and Lm using a method
as sensitive and specific as FSIS’. Products could be shipped into commerce after test
46
results on each individual lot are received while the establishment is validating its
process because the products would be considered to meet the definition of RTE in 9
CFR 430.1 (that it is in a form that is edible without additional preparation to achieve
food safety). The establishment could also choose to produce and sample experimental
products that are not offered for sale, are not introduced into commerce, and are not
given away to potential customers. Because experimental products are not for
introduction into commerce, they are not subject to inspection, are not eligible to bear
the mark of inspection, and do not need to be produced in accordance with the meat
and poultry products inspection regulations (9 CFR Part 300).

It would not be appropriate to use microbiological data gathered as part of in-plant

validation data to support large differences between a journal article and an
establishment’s process, rather a new challenge study would likely be needed to
support the unique combination of parameters. What is considered a large difference
will depend on factors such as the parameter (e.g., temperature, pH, water activity) and
the sensitivity of the scale for measurement (e.g., a 0.1 difference in temperature may
not have as big of an impact as a 0.1 difference in water activity). For example, if an
establishment producing a fermented meat product identifies a journal article that
matches its process, but it intends to ferment to a higher pH (e.g., 5.3 instead of 4.8 as
shown in the support) it would not be appropriate to rely on in-plant microbiological data
to support the differences. In this example, the establishment should conduct a
challenge study to support its unique combination of parameters. While the challenge
study is being conducted, products could be shipped into commerce during the 90 day
initial validation period if each individual lot of product is tested as described in 2. above
because the products would be considered to meet the definition of RTE in 9 CFR 430.1
(that it is in a form that is edible without additional preparation to achieve food safety).

47
Appendix 5: Scientific Support Available for Shelf-Stability

Establishments typically need different scientific support to support using either water
activity alone or a combination of pH and water activity to achieve shelf-stability than the
support used to achieve lethality. This is because to support shelf-stability,
establishments must have scientific support that these parameters allow no outgrowth
of S. aureus in the product during storage (9 CFR 417.5(a)(1)):

• The Food Standards and Labeling Policy Book available at:

https://www.fsis.usda.gov/guidelines/2005-0003 contains critical operational
parameters such as certain levels of MPR, pH, water activity, and brine
concentration for shelf-stable sausages or Lebanon bologna. Establishments can
(but are not required to) use the criteria as support for shelf-stability because these
criteria were based on ensuring no S. aureus growth would occur. Establishments
that use other criteria may still need to monitor the MPR as part of in-plant validation
and on-going verification activities for standard of identity purposes (e.g., salami
must have an MPR of 1.9:1 or less) to not be considered false and misleading (9
CFR 317.8 or 9 CFR 381.129).

NOTE: The regulatory standards of identity, 9 CFR 319.106, for country ham and
dry cured ham do not ensure or support the product is shelf-stable. Rather, the
criteria in the standard of identity were designed to ensure that the finished products
have quality characteristics associated with hams and pork shoulders (42 FR 3299).

• The FSIS Guideline for Meat and Poultry Jerky Produced by Small and Very Small
Establishments contains guidance on water activity parameters that support shelf-
stability. Although that guideline is about jerky, the water activity parameters can be
applied to other shelf-stable meat and poultry products.

• Research (Tilkens et al., 2015) supports that a combination of ≤ pH 5.1 and water
activity ≤ 0.96 in RTE snack sausages containing salt, dextrose, sodium nitrite, and
sodium erythorbate does not support the growth of S. aureus and thus could be
considered shelf-stable for this pathogen under anaerobic conditions (e.g., packaged
under vacuum). It also supports a combination of pH ≤ 5.1 and water activity ≤ 0.92
does not support the growth of S. aureus when products are stored aerobically
although mold grew under these conditions in the study; therefore, establishments
using these parameters should take additional measures to address mold growth.

• The University of Wisconsin Shelf-Stability Predictor available at:

https://meathaccp.wisc.edu/ST_calc.html provides accurate predictions for S. aureus
growth. It can be used alone as supporting documentation to support shelf-stability,
provided that the establishment’s product is like those the model was developed for
(Borneman et al., 2009; available on the shelf-stability predictor website).

48
Appendix 6: Critical Operational Parameters for Fermentation

During fermentation, bacteria consume available carbohydrates (e.g., dextrose or

sucrose) and moisture in the meat or poultry to produce organic acids (e.g., lactic acid
and citric acid) among other compounds to lower the pH and reduce the amount of
available water (reduce the water activity) in the meat. In meat or poultry, it is the lactic
acid that is primarily responsible for the reduction in pH (Benito, 2007; Ortiz, 2014). It is
important to note that even with fermentation that results in lactic acid production
and reduced pH, fermentation itself is not a particularly effective lethality
treatment and pathogens such as Salmonella, STEC, and Lm can survive (Faith et
al., 1997; Faith et al., 1998a; Faith et al., 1998b; Hussein et al., 2022; Ihnot et al., 1998).
One reason is because bacteria become tolerant to the acid produced during the
fermentation process particularly if it is a slow process (Leyer et al., 1993; Chikthimmah
and Knabel, 2001; Theron, 2007). The pathogens that do survive fermentation also
become tolerant to other treatments like heat that would otherwise be lethal in a typical
processed meat or poultry product (Brul, 1999).

Establishments must ensure that critical operational parameters within the scientific
support closely match the establishment’s actual process or provide justification for any
differences to support the decisions in its hazard analysis (9 CFR 417.5(a)(1)).
Examples of critical operational parameters for the reduction of Salmonella, STEC, and
Lm during fermentation include:

• Fermentation temperature, target pH, time to reach target pH, and smoke (if
used).
• Type and use of starter cultures.
• Product characteristics: Casing diameter and shape and product formulation
including salt, sugar (type and level), and use of nitrite or nitrate.

In addition to these critical operational parameters during fermentation, how the raw
batter is handled prior to fermentation can also have an impact on the effectiveness of
the step. For example, research showed that raw batter conditioned by tempering at
55.4°F (13°C) for 2 hours, freezing, and thawing prior to stuffing and fermentation
resulted in greater reductions in E. coli O157:H7 during fermentation than raw batter
that was either refrigerated or frozen and thawed (but not tempered first) prior to stuffing
and fermentation (Faith et al., 1998).

Specific considerations for several critical operational parameters for the reduction of
Salmonella, STEC, and Lm as related to fermented products are outlined below.

Fermentation Temperature, Target pH, Time to Reach Target pH, and Smoke (if
used)

The establishment must be fermenting its product to the pH that is recommended in the
supporting documentation or provide justification for any differences to support the
decisions in its hazard analysis (9 CFR 417.5(a)(1)). Temperature impacts the final pH
achieved during fermentation as well as the amount of time it takes to reach the target
pH. In general, the higher the fermentation process temperature and humidity, the
faster the fermentation (fermentation temperatures are typically no higher than 110°F).
49
However, the fermentation temperature should be at the optimum growth temperature of
the added starter culture. The final pH will be affected by the added carbohydrate, the
heating temperature after fermentation, and the drying conditions.

There are two key target pH values during fermentation:

• pH at which S. aureus growth is controlled (pH ≤ 5.3).

• Final product pH that contributes to lethality of Salmonella, STEC, and Lm

(which will vary depending on the process and supporting documentation).

In addition to the target pH level itself, the time it takes the product to reach the desired
pH is also important. For S. aureus control, it is important that the establishment monitor
the time it takes the product to reach pH of 5.3. By monitoring the product pH and time it
takes to reach the desired pH, the establishment can ensure its process is within an
acceptable number of degree-hours and, if applicable, achieves a secondary pH in an
amount of time consistent with the supporting documentation. If product takes longer to
achieve the secondary pH than occurred in the supporting documentation, the process
may not be as effective because it could allow time for the pathogens to adapt to the
acidic conditions. In general, the lower the final pH and the faster it is achieved the
greater reductions in pathogens such as Salmonella (Porto-Fett et al., 2008) although
as indicated on the previous page, fermentation itself is not a particularly effective
lethality treatment.

Added smoke will sometimes inhibit fermentation at the product surface, but the extent
to which this occurs will depend upon the product diameter.

NOTE: For those processes where fermentation is followed by cooking, FSIS does not
object to an establishment using the worst-case chamber temperature (110 °F) to set
the maximum allowable degree-hours and demonstrating 1) the total fermentation time
plus cooking is 18 hours or less (maximum allowable degree-hours at 110 °F) and 2)
the pH is 5.3 or less at the end of cooking.

Type and Use of Starter Cultures - Today, most fermented meat processors either
add lactic acid starter cultures and/or harmless staphylococci to the raw meat mix. The
composition of the starter culture (i.e., the strains) used in the product must be similar in
composition to that used in the supporting documentation, to ensure that fermentation is
achieved, and the rate of pH drop is as expected. If the starter culture is different the
establishment must provide justification for why the different starter culture would be
equally effective in order to support the decisions in its hazard analysis (9 CFR
417.5(a)(1)). The starter culture should be formulated to ensure microbial dominance of
fermentation strains over any potential pathogens and to inhibit potential S. aureus
growth during fermentation. In addition, the starter culture used for fermentation can
affect whether bacteriocins are produced and the type of bacteriocins produced, which
can affect the level of reduction for bacterial pathogens. For example, pediocin-
producing Pediococcus acidilactici are more effective than non-pediocin-producing
Pediococcus acidilactici at reducing Lm in chicken and turkey summer sausage
(Baccus-Taylor et al., 1993; Luchansky et al., 1992). It is also important that the starter
culture is well-dispersed throughout the raw batter.

50
NOTE: An alternative to commercial starter cultures for reducing the pH in sausage
batters is direct acidulation by the addition of organic acids like lactic, citric, or
glucono-delta-lactone (American Meat Institute, 1997). This guideline does not address
the use of direct acidulation.

Why is Using a Starter Culture Important?

If a starter culture and fermented sugars are not used, pathogenic bacteria will not be
reduced in the product during fermentation (Calicioglu et al., 2002; Smith and Palumbo,
1978; Smith and Palumbo, 1983). The use of a starter culture (e.g., lactic acid bacteria)
and fermentable sugars added to the initial fermented sausage mix assures microbial
dominance over the potential pathogenic microorganisms. This means that in the
absence of a starter culture Salmonella, STEC, and other undesirable microorganisms
will instantly become the dominant microflora. To prevent pathogen dominance,
establishments should use a starter culture to prevent or severely limit the growth of the
bacterial pathogens of public health concern during the initial phase of the fermentation
step of dry/semi-dry fermented sausages. The following factors are important to ensure
food safety:

• Microbial competition.

• Production of lactic acid which lowers the product pH and inhibits the
growth of pathogenic bacteria.

• Possible production of bacteriocins, which inhibit or reduce pathogenic

microorganisms.

As described in Good Manufacturing Practices for Fermented Dry and Semi-Dry

Sausage Products, there are two general ways in which lactic acid-forming bacteria for
fermentation may be incorporated into the raw batter:

1. The preferred and most reliable method is to use a commercially prepared

culture which is handled and used as prescribed by the manufacturer.

2. A less reliable procedure is the use of a portion of a previously fermented and

controlled mother batch. Because this method is less precise than using a
commercial culture, it is important that the inoculum derived from the mother
batch be composed of a vigorous culture
capable of producing a rapid pH decline.
Key Point
In addition to the concerns, identified in Good A starter culture helps ensure
Manufacturing Practices for Fermented Dry and that the fermentation is
Semi-Dry Sausage Products, using a mother batch vigorous enough to lower the
for fermentation is a less reliable procedure because
pH to 5.3 or less rapidly.
it could lead to recycling of spoilage organisms and
pathogens. Another concern with the use of a
mother batch is the inconsistency in the starter
culture. For these reasons, FSIS does not recommend this practice. If an establishment
uses a mother batch, it must address the potential for recycling of spoilage organisms
51
and pathogens in its hazard analysis as well as the consistency of the starter culture (9
CFR 417.2(a)(1)). Establishments may consider characterizing the type and level of
bacteria in the mother batch to support their decisions (9 CFR 417.5(a)(1). It is also
particularly important the establishment address the potential for S. aureus growth
during the fermentation and measure the degree-hours to ensure the pH is reduced to ≤
pH 5.3 within an acceptable number of degree-hours at a given fermentation
temperature as described on page 37.

As indicated in the GMP document, a third method, used historically, relied on lactic
acid bacteria which naturally occur in fresh meat to initiate the fermentation. While this
practice had been used in the past and was the original art of making fermented
sausage; the method is highly unreliable and should not be used.

Product Characteristics: Casing diameter and shape and product formulation

including salt, sugar (type and level), and use of nitrite or nitrate.

The following product characteristics are important critical operational parameters to

ensure the fermentation step is effective:

The casing diameter will impact the fermentation rate and final pH by affecting heat
penetration and moisture migration in, and then out, of the sausage chub. Generally,
large diameter products ferment slower due to slower heat transfer and they take longer
to reach the desired final pH.

Product formulation including salt, sugar (both the type of sugar and amount)
and use of nitrite or nitrate plays a role in the fermentation process and may also
affect microbial tolerance to acid or heat. The establishment should understand the
critical operational parameters associated with the product formulation (e.g., % salt,
sugar type and level, moisture level, level of nitrite or nitrate or any other preservatives,
and % fat) and should ensure that the materials used in the supporting documentation is
like their product with respect to those critical operational parameters and ingredients.
Reduced salt was a potential contributing factor in a 2021 salmonellosis outbreak
associated with Italian-style meats (FSIS, 2022).

Although typically thought of for Clostridial control, nitrite is also a hurdle which inhibits
Salmonella (Honikel, 2010). According to Honikel, 2010, “fermentation can be produced
with only salt, but there is a greater microbial risk if no nitrite is used.” Nitrite alone may
be used, or it may be used in combination with nitrate, which is often added as a
reservoir for nitrite in long-term processing. Regardless, establishments must ensure
the same amount of nitrite or nitrate, as well as any cure accelerators, such as
ascorbate or erythorbate, is used as in their scientific support or provide justification for
any differences to support the decisions in its hazard analysis (9 CFR 417.5(a)(1)).
Establishments must also ensure the levels of sodium nitrite (whether from a natural or
synthetic source) and other restricted ingredients are safe and suitable according to
FSIS Directive 7120.1, Safe and Suitable Ingredients Used in the Production of Meat
and Poultry Products and 9 CFR 424.21(c)).

If establishments are using natural sources of sodium nitrite, FSIS recommends that
establishments use natural sources of sodium nitrite with known concentrations of
nitrite. By knowing the concentration of nitrite, establishments can ensure they are
52
adding the same amount as used in the scientific support. Because natural sources of
nitrite are not currently approved for use in 9 CFR 424.21(c) as curing agents, products
that are required to contain curing agents and cure accelerators as part of a standard of
identity in 9 CFR 319 or 9 CFR 317.17(b), but instead are formulated with natural
sources of nitrite and ascorbate, must be labeled as “uncured” under 9 CFR 319.2.
Also, the label must contain the statement “no nitrates or nitrites added” (9 CFR 317.17)
that is qualified by the statement, “except for those naturally occurring in [name of
natural source of nitrite such as celery powder]” as to not be considered misbranded
due to false and misleading labeling under 9 CFR 317.8. Uncured products are also
required to bear the handling statement, “Not Preserved - Keep Refrigerated Below 40
°F. At All Times” as per 9 CFR 317.17(c)(2). However, this handling statement is not
required if an uncured product is processed to be shelf stable by fermenting or pickling
to pH of 4.6 or less or drying to a water activity of 0.92 or less. For more information see
FSIS’ Stabilization Guideline for Meat and Poultry Products (Revised Appendix B).

53
Question: Why are there so many critical operational parameters during
fermentation? If I meet the degree-hours isn’t that enough to show biological
hazards are addressed?

Answer: No, degree-hours are used to control the outgrowth of S. aureus. To reduce
levels of other pathogens of concern in fermented products, such as Salmonella, STEC,
and Lm, establishments often need to ferment these products to a lower pH. Other
parameters, such as fermentation temperature, time to final pH, and formulation, also play
a role in reducing other types of bacteria. All critical operational parameters within the
scientific support must closely match the establishment’s actual process or the
establishment must provide justification for any differences to support the decisions in its
hazard analysis (9 CFR 417.5(a)(1)).

Question: What if I meet the degree-hours and dry my product following one of the
methods in FSIS’s Trichinella Guideline and dry to a reduced water activity such as
to a water activity below 0.85, isn’t that enough to show biological hazards are
addressed?

Answer: No. Fermentation and drying alone are not particularly effective as lethality
treatments. When these steps are found to achieve sufficient reductions in pathogens,
there are a lot of critical operational parameters associated with both steps that need to be
implemented in the actual process consistent with the scientific support. It is not enough to
only meet degree-hours, follow the minimum number of drying days for eliminating
Trichinella, and achieve a final water activity for shelf-stability. Degree-hours are intended
to control the outgrowth of S. aureus. To reduce levels of other pathogens, such as
Salmonella, STEC, and Lm, products often need to be fermented to a lower pH than 5.3.
Also, the drying time and other factors are important to reducing bacteria during drying –
not just the final water activity. The minimum number of drying days for eliminating
Trichinella have not been validated to achieve any particular reductions in Salmonella and
products often have to be dried longer to get significant reductions in Salmonella. In
addition, smaller diameter products tend to dry faster than large diameter products, and the
shorter drying time is often not enough time to achieve the same reductions in bacteria as
larger diameter products even if it is enough time to achieve the targeted water activity
(DeSouza, 2018).

Here are some citations that support the need for other critical operational parameters
establishments should address during fermentation, in addition to degree-hours:

Blue Ribbon Task Force of the National Cattlemen’s Beef Association. May 1996.
Dry Fermented Sausage and E. coli O157:H7 (Research Report No. 11-316).
• Table 6 in the article shows that when large diameter (105 mm) salamis were
fermented at 90°F to a pH ≤ 4.6 and held, the average E. coli O157:H7 log
reduction was 4.72. In contrast, when the products were fermented at 90°F to a
pH ≥ 5.0 and held in the chamber at that temperature for 7 days, a 2.87-log
reduction was measured. The same findings, although less pronounced, were
observed for large diameter salamis fermented at 110°F to a pH ≤ 4.6 and held.

54
 The average E. coli O157:H7 log reduction was 6.42 as opposed to 6.03-log
reduction for products fermented to ≥ 5.0.

Porto-Fett, ACS, Hwang, C-A, Call, JE, Juneja, VK, Ingham, SC, Ingham, BH,
Luchansky, JB. 2008. Viability of Multi-Strain Mixtures of Listeria monocytogenes,
Salmonella Typhimurium, or Escherichia coli O157:H7 Inoculated into the Batter
or Onto the Surface of a Soudjouk-Style Semi-Dry Sausage. Food Microbiology.
25: 793-801.
• The authors fermented sausage with different levels of dextrose that resulted in a
noticeable difference in pH values between sausages (pH 5.27 when formulated
with 0.25% dextrose versus pH 4.81 when formulated with 0.60% dextrose).
• The authors reported the addition of a pedicococcal starter culture and added
dextrose decreased the pH to approximately 5.3 or 4.8 following fermentation
and drying, and the lower pH resulted in a quantifiable greater lethality.

These scientific support documents illustrate why meeting degree-hours is not enough
to show biological hazards (i.e., Salmonella, STEC, and Lm) are addressed by the
fermentation step.

For information related to fermentation deviations see Appendix 7.

Other critical operational parameters during fermentation that do not need to be

addressed

A high relative humidity (85-90%) is preferred during fermentation to keep the product
surface slightly moist (tacky) during fermentation and prior to subsequent drying. This
avoids premature and uneven drying at the surface and the humidity enhances the
fermentation by limiting undesirable evaporative cooling, increasing heat penetration,
and speeding the reduction in pH. For these reasons, establishments should try to
ensure that the relative humidity in the actual process is at least as high as the lower
end of the relative humidity range used in the supporting documentation. It is
recommended establishments determine the relative humidity levels in the actual
process during the initial set-up of the system as part of in-plant validation as well as
part of ongoing verification.

Establishments that are unable to determine the relative humidity level used in the
scientific support or that are not able to meet the same or higher level of relative
humidity from the support may also be able to use the implementation of other
parameters such as final fermentation pH and time to reach pH to demonstrate that
adequate relative humidity is present for the fermentation process. For example, The
Blue Ribbon Task Force document, commonly used as scientific support for fermented
sausage processes, does not report the level of relative humidity that was used. FSIS
would not object to establishments using this document as support and concluding the
relative humidity levels in the actual process are sufficient if the final fermentation pH
and time to reach pH are consistent with the levels reported in The Blue Ribbon Task
Force document. Indications of insufficient relative humidity include partial or
inconsistent fermentation (for more information see Appendix 7: Fermentation
Deviations) or indications of case hardening (e.g., demonstrated by a lack of loss of
weight during drying). If possible, FSIS recommends establishments still monitor the
relative humidity levels as part of in-plant validation and ongoing verification so that
55
there is data available on typical levels.

The casing type influences moisture exchange. Products with impermeable, semi-
permeable, or permeable casings exchange moisture with the environment differently
and can influence the rate of product acidification, the penetration of heat into the
interior of the product, and the maximum internal temperature reached by the product.
However, several studies have found no differences in reductions for pork salami
fermented and dried in natural, collagen, or fibrous casings (DeSouza, 2018; McKinney
et al., 2019). Therefore, establishments using scientific support conducted with a
product fermented in one type of casing may be able to support applying that research
to a product fermented in a different type of casing.

56
Appendix 7: Fermentation Deviations

If the fermentation to a desired pH does not occur within the normal time, it can be due
to a variety of reasons. If a fermentation problem does occur, it is often the result of:

• No fermentation (the pH does not change from its initial value of pH 5.6-6.0).

• Partial fermentation (the pH drops slightly to pH 5.4-5.6).

• Inconsistent fermentation (variation in fermentation activity from piece to piece or

from location to location).

Below are some typical causes for inadequate or inconsistent meat and poultry product
fermentation.

No Fermentation

• No starter culture added.

• No fermentable sugars added.
• Excessive salt added.
• Culture was thawed and refrozen.
• Culture premixed with cure, salt, chemicals.
• Antimicrobial agents added to formulation.
• Antibiotic residues in raw meat.

Inconsistent or Partial Fermentation

• Inadequate distribution of starter culture.

• Insufficient fermentable sugars added.
• Fermentation temperature above or below the optimal temperature for the starter
culture.
• Inconsistent internal product temperature or processing temperature or humidity.
• Reduced fermentative activity of the starter culture.
• Out of code product or improper stock rotation.
• Culture was temperature abused.
• Antimicrobial agents added to formulation.
• Antibiotic residues in raw meat.

One or more of these causes may occur during the fermentation of a meat and poultry
product, resulting in a fermentation deviation. Establishments should ensure that critical
operational parameters within the scientific support that should closely match the
establishment’s actual process.

Evaluation of Product Safety

Establishments must be able to ensure that no product that is injurious to health or

otherwise adulterated because of the deviation enters commerce, and to support its
product disposition decisions (9 CFR 417.3(a) and (b)). In the event of a fermentation
57
deviation, establishments may use pathogen modeling (DMRI Staphtox Predictor) or
testing to support product safety. Below are specific recommendations for either
approach.

Pathogen Modeling Following a Fermentation Deviation

Establishments may consider using the DMRI Staphtox Predictor (Version 1.0)
available at: http://dmripredict.dk/ to assess the likelihood of S. aureus growth and toxin
formation during a constant temperature fermentation deviation of sausages (water
activity > 0.96). As explained on page 46, this model has been validated. The seven
input variables and their ranges for entering information into the model are provided
below:

Input variables and ranges:

1. NaCl in product: 1.8 to 4.2% in steps of 0.1%.
2. KCl in product: 0 to 4.2% in steps of 0.1%.
3. Sodium nitrite in the recipe/ingoing: 0-150 ppm in steps of 10 ppm.
4. % water in product as determined through laboratory analysis: 62-78% in steps
of 1%.
5. Time: in hours.
6. pH: 4.6-6.0 in steps of 0.1 or pH change during fermentation of sausages.
7. Temperature: 15-40°C in steps of 1°C or time and temperature profile (maximum
500 data points or rows).

If modeling estimates < 3.0-log growth of S. aureus and no toxin formation, then
modeling is adequate to show that the process prevented enterotoxin formation.

If modeling estimates ≥ 3.0-log growth of S. aureus, then product should be tested

following the guidance below (even if the model predicts no toxin formation).
FSIS does not know of other validated pathogen modeling for estimating growth and
toxin formation during a constant temperature fermentation deviation.

Testing Following a Fermentation Deviation

FSIS recommends that establishments follow the recommendations in the Good

Manufacturing Practices for Ferment Dry and Semi-Dry Sausage Products for
evaluating product safety in the event of a fermentation deviation. Although the
guidance document recommends at least three samples be collected per batch, FSIS
recommends establishments use a statistically based sampling plan (e.g., ICSMF
sampling plans).

FSIS recommends establishments develop sampling plans that include:

• A case 11 or n=10 sampling plan be used in the event of a fermentation

deviation, although establishments can provide support for other sample sizes
(for more information on the ICMSF sampling plans, see page 120 of The FSIS
Guideline: Controlling Listeria monocytogenes in Post-Lethality Exposed RTE
Meat and Poultry Products).

58
 • If the product is not cooked, the establishment should:
o Test for the enterotoxin if any samples contain ≥ 1,000 CFU/g < 10,000
CFU/g of S. aureus.
o Reject the lot and condemn the product if any samples contain ≥ 10,000
CFU/g.

• If product has undergone either a low- temperature or high-temperature heat

step, establishments should test for the enterotoxin only as any vegetative cells
of S. aureus would likely be destroyed by the heat treatment.

FSIS does not recommend:

• Compositing of samples because the purpose of testing following a fermentation
deviation is to determine the level of S. aureus and compositing may dilute the
sample.

59
Appendix 8: Critical Operational Parameters for a Low-Temperature
Heat Step

To achieve adequate reductions of pathogens in fermented products, while still

maintaining the desired quality characteristics, a low-temperature heat step may be
added after the fermentation is complete. For example, a fermentation and drying
process for pepperoni achieved ≤ 2.0-log reduction of E. coli O157:H7; but, by adding a
low-temperature heat step after fermentation (128°F for an hour) they were able to
achieve a ≥ 5.0-log reduction of E. coli O157:H7 without changes to quality (Hinkens, et
al.,1996). Dry-curing beef strips, followed by a low-temperature heat step, followed by
drying has been found to result in a 5-log reduction of Salmonella in a salt-cured
basturma product (Genigeorgis and Lindroth, 1984).

Establishments must ensure that critical operational parameters within the scientific
support closely match the establishment’s actual process or provide justification for any
differences to support the decisions in its hazard analysis (9 CFR 417.5(a)(1)).
Examples of critical operational parameters for the reduction of Salmonella, STEC, and
Lm using a low-temperature heat step include:

• Time and temperature: Heating come-up time (CUT), hold time, and temperature
for low temperature heating step.
• Equipment used to generate heat.
• Product characteristics.

Time and Temperature: Heating Come-up Time, Hold Time, and Temperature for
Low-Temperature Heat Step

The temperature that the product is heated to, and the amount of time the product is
held at this temperature (hold time and temperature), are critical to ensuring that
adequate lethality is achieved. In addition to the hold time and temperature, the come-
up time, which is the time it takes the product to reach the target temperature for the
post-fermentation low-temperature heat step, may be important to ensure product
safety. Factors such as product diameter and relative humidity, affect heat transfer and
the amount of time it takes the product to reach the target temperature. It is important
for the establishment to understand how the actual temperature of the product, the
CUT, and the amount of time the product is held at the target temperature compared to
the supporting documentation. For example, if the CUT in the establishment’s process
is shorter than the time it takes in the study, the establishment’s process may result in a
lower level of pathogen reduction and not achieve product safety.

Equipment Used to Generate Heat

Differences in equipment (e.g., smokehouses and ovens) can influence the

effectiveness of the process and the speed of fermentation or acidification and heating.
For this reason, the establishment should gain an understanding of the pH and
temperature profile of the product throughout the process. In addition, seasonality of
atmospheric conditions, cold-spot determination, or heating consistency must be
understood to support monitoring and verification procedures and the frequencies at
which those procedures are monitored and verified (9 CFR 417.5(a)(2)).
60
Product Characteristics: Casing Diameter and Product Formulation
Product casing diameter is a critical operational parameter in fermented, dry and semi-
dry processes because it affects heat transfer. It is important that the diameter of the
product used in the establishment’s process is the same or smaller than that of the
product used in the supporting documentation. If the diameter of the establishment’s
product is larger than that of the product used in the supporting documentation, it is
possible that the product core will take longer to reach the desired temperature and pH,
and a lower level of pathogen reduction would be achieved.

See page 52 information on product formulation.

Other critical operational parameters of interest during low temperature heating

that do not need to be addressed

Relative Humidity During the Heating Step

FSIS considers relative humidity to be inherently maintained and, therefore, does not
need to be addressed, for products that are cooked in a casing (including a natural
casing) even for products that are cooked and then dried. For more information on
relative humidity during heating see page 25 of the FSIS Cooking Guideline for Meat
and Poultry Products (Revised Appendix A).

See page 56 for information on casing type.

61
Appendix 9: Critical Operational Parameters for Salt-Curing and
Equalization

Dry-curing is the process of adding salt, sugar, nitrite, or nitrate by dredging the meat
in the cure (known as the salt box method) or by applying a pre-weighed cure mixture
to the surface of the product. FSIS recommends applying a pre-weighed cure mixture
over the entire surface, instead of the salt box method, because this process ensures
the same amount of ingredients are added in the actual process as was used in the
supporting documentation. During this process, the salt, sugar, nitrite, and/or nitrate
begin to migrate throughout the meat tissue. At the same time, moisture from the meat
or poultry tissue is extracted by the salt surrounding the meat.

For dry-curing to be effective as a food safety measure, time needs to be given to allow
equalization, which is the process where the cure mixture migrates throughout the
meat tissue.

NOTE: High-salt tolerant starter cultures may be added directly to the brine or dry rub
used for curing of the whole muscle to provide consistent flavor and improved color.
Starter cultures may consist of non-pathogenic Staphylococci or Lactobacilli strains or a
combination of both. The use of starter cultures on whole-muscle cuts is not addressed
in more detail in this document since they are usually used for color and flavor
development and do not generally impact product safety.

Establishments must ensure that critical operational parameters within the scientific
support closely match the establishment’s actual process or provide justification for any
differences to support the decisions in its hazard analysis (9 CFR 417.5(a)(1)).
Examples of critical operational parameters for the reduction of Salmonella, STEC, and
Lm during the curing or formulation step include:

• Curing temperature.
• Curing time.
• Salt coverage of exposed muscle tissue.
• Product characteristics (e.g., product size and formulation, including salt
concentration).

Curing Temperature

The curing temperature (typically ambient curing room temperature) is a critical

operational parameter as it impacts the salt migration throughout the product.
Specifically, a lower temperature during curing will slow salt migration throughout the
product impacting the final brine concentration and water activity at the end of the curing
step. During curing, establishments must ensure that growth of pathogens does not
occur to significant levels (9 CFR 417.2(a)(1)). Storing at refrigeration temperatures
(e.g., by storing below 45°F or 41°F according to the Tompkin paper) is one way to
support decisions at that step (9 CFR 417.5(a)(1)).

62
Curing Time

Since cure migration occurs throughout the entire time the product is being cured, the
length of curing time is critical to ensure that a sufficient cure penetration and adequate
brine concentration are achieved throughout the product.

Salt Coverage
Salt coverage is also critical as the amount of coverage, the amount of salt, and type of
salt (discussed below) will ultimately impact the brine concentration achieved after the
equilibration step. The coverage may be described in the supporting documentation
either: (1) through complete coverage of the whole muscle cut or (2) the number of
pounds per product or per surface area.
Product Characteristics: Product Size and Formulation
Product size impacts the surface area, and therefore, the amount of cure needed.
Product size also impacts curing and salt equalization times. A greater product surface
area requires longer curing times to ensure sufficient cure penetration throughout the
product.

Product formulation that impacts lethality includes the amount and use of:

Salt
Establishments must use the same or higher amount or concentration of salt as
used in the supporting documentation to ensure the brine concentration and
water activity throughout the product after equalization is like what was achieved
in the support or provide justification for any differences to support the decisions
in its hazard analysis (9 CFR 417.5(a)(1)). Establishments should use salt by
weight rather than volume because the size of the salt grains make a big
difference. Fine crystalline salt takes up less space (volume) and will weigh more
than an equal volume of coarse granular salt. In addition to the concentration,
uniform salting over the entire surface is critical to inhibit pathogens and
spoilage microorganisms. Moreover, some products may be re-salted several
times after the first salting step as the salt migrates into the tissue.
Sodium Nitrite (NaNO2)

Nitrite is an important component during curing because it, in combination with

salt, completely inhibits the growth of C. botulinum. Sodium nitrite, in combination
with salt, also greatly inhibits the growth of C. perfringens, and slows the growth
of many other pathogenic bacteria such as Salmonella and Lm (Buchanan et al.,
1989; Honikel, 2010; Sindelar, 2012). The amount of nitrite used in dry-cured
products, such as coppa, country ham, prosciutto, and others, can be up to 625
ppm ingoing nitrite for dry-cured products and is based on the green weight of
the meat in the product formulation. As discussed in Appendix 2, 50 ppm sodium
nitrite is considered sufficient to ensure C. botulinum will not grow (Reynolds et
al., 2006; Johnston et al., 1969; and Tompkin, 1976). The resting or equalization

63
 phase itself is also a critical step for inhibiting C. botulinum growth in dry-cured
hams (Merialdi et al., 2016).

Sodium Nitrate (NaNO3)

Sodium nitrate is used as a source of nitrite. If nitrate is used as the curing agent,
the conversion (reduction) of nitrate to nitrite by bacteria in the meat or poultry or
by a nitrate-reducing starter culture is a necessary step. The amount of nitrate
that is reduced to nitrite is dependent upon the numbers of nitrate-reducing
bacteria and several environmental conditions, such as temperature, moisture
content, salt content, and pH. For this reason, the conversion rate and
subsequent amount of nitrite that is formed is difficult to control. The poor control
associated with the reduction of nitrate to nitrite, coupled with the fact that most
processors today demand faster curing methods, has led to the diminished use
of nitrate in meat and poultry products. FSIS recommends the use of nitrite
instead of nitrate.

Equalization

After curing, products go through an equalization step (typically ≤ 45°F) to ensure salt
migrates throughout the product and a sufficient brine concentration and water activity is
achieved. Equalization is necessary to prevent the growth of S. aureus and other
pathogens when temperature is increased during the drying step; this allows for
sufficient salt penetration and equilibrium. Equalization often takes many weeks to
achieve uniform salt distribution to a level greater than 10%.

Establishments must ensure that critical operational parameters within the scientific
support closely match the establishment’s actual process or provide justification for any
differences to support the decisions in its hazard analysis (9 CFR 417.5(a)(1)).
Examples of critical operational parameters for the reduction of Salmonella, STEC, and
Lm during an equalization step include:

• Equalization temperature.
• Equalization time.
• Brine concentration and water activity after equalization.
• Product size (diameter or thickness).

Equalization Temperature

The equalization temperature is a critical operational parameter as it impacts the salt

migration throughout the product. Specifically, a lower temperature during equalization
will slow salt migration throughout the product impacting the final brine concentration
and water activity. During equalization, establishments must ensure that growth of
pathogens does not occur to significant levels (9 CFR 417.2(a)(1)). Storing at
refrigeration temperatures (e.g., by storing below 45°F or 41°F according to the
Tompkin paper) is one way to support decisions at that step (9 CFR 417.5(a)(1)).

64
Equalization Time

The equalization time is critical to ensuring sufficient brine

concentration and water activity at the end of the process. If
Key Point
a process uses a shorter equalization time, then the final
brine concentration and water activity may not be sufficient If a process uses a shorter
to prevent S. aureus growth when product begins drying at equalization time than that in
a higher temperature. the supporting documentation,
then the final brine
Brine Concentration and Water Activity after concentration and water activity
Equalization
may not be sufficient to prevent
Establishments must ensure that the product conditions S. aureus growth when product
during curing and equalization are such that the product has begins drying at a higher
a high enough brine concentration and low enough water temperature.
activity at the end of the equalization stage prior to drying to
prevent S. aureus outgrowth during drying according to the
scientific support. If the establishment is not using brine concentration and water activity
as controls measured at the end of the equalization stage to prevent S. aureus
outgrowth during drying then the establishment must provide other support for the
decisions in its hazard analysis (9 CFR 417.5(a)(1)). Staphylococcal enterotoxin
production is inhibited at brine concentrations above 10% (Jay, 2000; Tatini et al.,
1976). Achieving a high enough brine concentration and low enough water activity will
ensure that when the product moves into the drying step where temperatures are
elevated that S. aureus growth is prevented or limited (≤ 2.0-log CFU/g). If the
establishment is elevating temperatures during equalization then it must support the
product has a high enough brine concentration and low enough water activity at the end
of curing to prevent S. aureus outgrowth during equalization and drying according to the
scientific support or must provide other support for the decisions in its hazard analysis
(9 CFR 417.5(a)(1)).

Product Size

Product size is also a critical factor. As product size increases, so does the equalization
time since it will take longer for moisture to transfer from the center of the product to the
surface and take longer to evaporate.

Other critical operational parameters during salt-curing and equalization that do

not need to be addressed

Relative humidity
Relative humidity in the curing environment impacts the amount of time it takes the cure
to migrate throughout the product. If the relative humidity is too high, the curing process
will be slowed, resulting in insufficient brine concentration/water activity throughout the
product. Therefore, if the brine concentration/water activity at the end of
curing/equalization meets the desired target, the establishment can support relative
humidity does not need to be addressed.

65
Air Flow

Along with relative humidity, air flow impacts the moisture loss from the surface of the
product. If the air flow is too fast, the product may dry out faster. If air flow is too slow,
the moisture loss may also be too slow when compared with the conditions that were
studied. Therefore, if the product weight loss meets quality/yield targets, the
establishment can support air flow does not need to be addressed.
Drying

After dry-curing and equalization, products are typically dried above refrigeration
temperatures so that additional water (moisture) is removed from the product. Salt-
cured products are dried to meet a water activity level sufficient to achieve shelf-stability
by preventing the growth of microorganisms (e.g., water activity ≤ 0.85), especially
toxigenic microorganisms such as S. aureus. Drying to an intermediate or low water
activity also may achieve additional reductions in Salmonella, STEC (in products
containing beef), and Lm. For an explanation of the critical operational parameters
associated with drying, see Appendix 11.

66
Appendix 10: Critical Operational Parameters for Seasoning/Marination
of Dried Products

During seasoning and marination salt is added to strips of meat as a dip or as a mixture
with other ingredients (e.g., spices, sugar, pepper, or sodium nitrite). The addition of
ingredients during formulation and the method of application of antimicrobials (if used,
for example, to add additional lethality) can impact their effectiveness at the time of
application but also the effectiveness of the subsequent steps including drying.

Establishments must ensure that critical operational parameters within the scientific
support closely match the establishment’s actual process or provide justification for any
differences to support the decisions in its hazard analysis (9 CFR 417.5(a)(1)).
Therefore, the following critical operational parameters associated with seasoning and
marination to reduce Salmonella, STEC, and Lm should be considered:

• Product formulation.
• Antimicrobial application (e.g., concentration, pH, coverage, contact time).

Product Formulation

Product formulation plays a role as the addition of ingredients (such as spices) may
impart some antimicrobial activity. Product formulation also may affect microbial
tolerance to acid or heat. The establishment should understand the critical operational
parameters associated with the product formulation (e.g., % salt, moisture level, level of
nitrite or any other preservatives, and % fat) and must ensure that the material used in
the supporting documentation is like their product with respect to those critical
operational parameters or provide justification for any differences to support the
decisions in its hazard analysis (9 CFR 417.5(a)(1)).

Antimicrobial Application

Some dried products, such as biltong, contain a marination step with acids such as
vinegar (acetic acid) that contribute to the overall reductions of pathogens. Critical
operational parameters for antimicrobial application include the pH, temperature,
pressure or flow rate (if applicable), coverage, and contact time. Establishments should
be aware that it is not appropriate to add up the results of two separate studies
conducted for the same type of intervention (such as two acid dips) because the second
time the intervention is used it will likely be less effective. This is because any bacteria
that survive the first treatment are likely to be more tolerant to the second treatment.

Drying

After seasoning and marination, products are typically dried at elevated room
temperature so that additional water (moisture) is removed from the product. Products
are dried to meet a water activity level sufficient to achieve shelf-stability by preventing
the growth of microorganisms (e.g., water activity ≤ 0.85), especially toxigenic
microorganisms such as S. aureus. Drying to an intermediate or low water activity also
may achieve additional reductions in Salmonella, STEC (in products containing beef),
and Lm.
67
Appendix 11: Critical Operational Parameters for Drying

Drying (sometimes called maturation for fermented products) is the process during
which water (moisture) is removed from the product. Shelf-stable products are dried to
meet a water activity level sufficient to prevent the growth of microorganisms, especially
toxigenic microorganisms such as S. aureus. Drying is typically performed in a drying
room to control the critical operational parameters of the process. Drying to an
intermediate or low water activity also may achieve additional reductions in Salmonella,
STEC (in products containing beef), and Lm (although Lm is more tolerant to drying
than Salmonella). The lower pH achieved by fermentation aids in drying since the meat
proteins are less able to bind water under acidic conditions.

Establishments must ensure that critical operational parameters within the scientific
support closely match the establishment’s actual process or provide justification for any
differences to support the decisions in its hazard analysis (9 CFR 417.5(a)(1)).
Examples of critical operational parameters for reduction of Salmonella, STEC, and Lm
during a drying step include:

• Drying room temperature.

• Drying time.
• Target water activity.
• Product characteristics.

Drying Room Temperature

It is critical that establishments use the same drying room temperature range that is
used in the supporting documentation to achieve similar reductions in levels of
pathogens of concern. As temperature increases the rate of drying will increase.
Although this can result in similar reductions, care must be taken to ensure that meat
surfaces do not become too dry while there is still a high moisture content inside the
meat pieces (a phenomenon known as case hardening). Dry surfaces inhibit the
further evaporation of moisture, which may result in products not uniformly dried and in
microbiological spoilage starting from the areas where the moisture content remains too
high (FAO, 1990). Research has also shown that storage under vacuum for extended
time following fermentation and drying is more effective when done under room
temperature than refrigeration (Ingham et al., 2004).

Drying Time

Establishments also need to ensure the drying time is consistent with the time used in
the scientific supporting documents to reach the desired water activity. If product takes
less time to dry, then the drying process will not be as effective as was shown in the
support as the longer drying time can increase stress on the bacteria (Gunvig et al.,
2017; Mutz et al., 2019). One main issue is that smaller diameter products tend to dry
faster than large diameter products, and the shorter drying time is often not enough time
to achieve the same reductions in bacteria as larger diameter products even if it is
enough time to achieve the targeted water activity (DeSouza, 2018). The use of a
shortened drying time for a smaller diameter product was a potential contributing factor
in a 2021 salmonellosis outbreak associated with salami sticks (FSIS, 2022). If an
68
establishment is following a scientific support document performed with one size
diameter product and they would like to produce a smaller diameter product, the
establishment may consider holding the smaller diameter product for the total drying
time needed for the larger diameter product. Ultimately, establishments need to
determine the impact any differences between the support and the establishment’s
actual process would have on the expected reduction. If differences are large, a
challenge study may be needed (for more information see Appendix 15).

Target Water Activity

The target water activity must be the same at the end of the drying time as what was
used in the supporting documentation or the establishment must provide justification for
any differences to support the decisions in its hazard analysis (9 CFR 417.5(a)(1)).
Once the target water activity is reached within the desired time, establishments may
continue drying product for a longer amount of time than in the scientific support to
achieve a lower water activity (e.g., for shelf-stability). Establishments should not
assume because a product is dried to a lower water activity within the same amount of
time as the scientific support that similar or higher reductions in pathogen levels will be
achieved. For example, when chorizo sausages were dried to target water activity levels
and held, the optimal death rate of Salmonella and E. coli O157:H7 occurred at a level
above that of the lowest water activity tested and that “sweet spot” varied by bacteria
(Hew et al., 2006).

NOTE: Establishments must follow the procedure used to measure water activity in the
scientific support or, if using a different procedure, must provide support for the
procedure as required by 9 CFR 417.5(a)(2). For larger products, samples should be
cut into small pieces. For most products, collecting representative samples from
external and internal portions of the product is recommended. However, for thicker non-
intact products (e.g., thicker biltong strips made from vacuum-tumbled meat), it may be
more appropriate to collect internal samples that would represent a worst-case scenario
(Karolenko, 2020).

Product Characteristics: Product Diameter and Product Composition

Product diameter (thickness) impacts drying time with thicker or larger diameter
products taking longer to dry. See drying time discussion above for further
considerations.

Product Composition

pH, amount of fat, and particle size also impact the drying rate (Toldra, 2002). Lowered
pH increases the drying rate in fermented meats due to the decrease in water-holding
capacity when the pH is around the isoelectric point of the proteins (Acton and Keller,
1974). Higher fat may result in reduced effectiveness of fermentation and drying (Faith
et al., 1998b). Establishments must ensure its product composition is like that of the
product studied in the supporting documentation or provide justification for any
differences to support the decisions in its hazard analysis (9 CFR 417.5(a)(1)).

For fermented products, the pH at the end of drying may also be a critical operational
parameter. The pH most often stays the same or goes up slightly. pH increases occur
69
likely as a result of proteolysis and the production of non-protein nitrogen-containing
compounds. In a few cases, pH decreases occur likely as a result of some fermentable
sugar remaining and the specific starter culture still being active after fermentation (for
example, if there is no low-temperature heat step) (Gunvig, 2016). A reduction in pH
after drying has been associated with greater reductions in pathogens (Deibel
Laboratories/Chr. Hansen, 2017).

Question: Why are there so many critical operational parameters during

drying? If at the end of my study a certain water activity is reached, why isn’t it
enough just to know my product gets the same water activity?

Answer: Because smaller diameter products tend to dry faster than large diameter
products, and the shorter drying time is often not enough time to achieve the same
reductions in bacteria in these products as in larger diameter products, even if it is
enough time to achieve the targeted water activity (DeSouza, 2018). In addition to
that drying time, drying temperature also plays a role in reducing bacteria.

Here are some citations that support there are other critical operational parameters in
addition to water activity at the end of drying:

DeSouza, J.D., Ahmed, R., Strange, P., Barbut, S., and Balamurugan, S. 2018.
Effect of caliber size and fat level on the inactivation of E. coli O157:H7 in dry
fermented sausages. Internal Journal of Food Microbiology. 266: 167-172.
• The authors found that when large diameter sausages reached a water activity of
0.85 after approximately 55 days a 5.0-log reduction in E. coli O157:H7 had been
achieved. However, when small diameter sausages with the same formulation
and fermentation and drying schedule reached 0.85 after approximately 17 days,
only a 2.0-3.0-log reduction in E. coli O157:H7 was achieved. To get to a 5.0-log
reduction, the smaller diameter products had to be dried longer and to a lower
water activity. If water activity is considered the only critical operational
parameter, then insufficient reductions may be achieved at smaller diameters.

Porto-Fett, ACS, Hwang, C-A, Call, JE, Juneja, VK, Ingham, SC, Ingham, BH,
Luchansky, JB. 2008. Viability of multi-strain mixtures of Listeria monocytogenes,
Salmonella typhimurium, or Escherichia coli O157:H7 inoculated into the batter or
onto the surface of a soudjouk-style semi-dry sausage. Food Microbiology. 25:
793-801.
• The authors found that as soudjouk-style sausages (fermented and dried) were
stored, Lm numbers decreased over time.
Gunvig, A., Borggaard, C.., Hansen, F., Hansen, T.B., and S. Aabo. 2016.
ConFerm – A tool to predict the reduction of pathogens during the production of
fermented and matured sausages. Food Control: 67: 9-17.
• The authors found processing time to be a significant variable for predicting Lm
reductions in fermented sausages.

70
These scientific support documents illustrate why meeting the water activity at the end
of drying is not enough to show biological hazards (i.e., Salmonella, STEC, and Lm) are
addressed by the fermentation step.

Other critical operational parameters during drying that do not need to be

addressed

Casing Type (for fermented products)

Pore diameter of casing, as well as casing type, may impact the drying rate because it
influences moisture exchange (Toldra, 2002). However, as described on page 56,
several studies have found no differences in reductions for pork salami fermented and
dried in natural, collagen, or fibrous casings (DeSouza, 2018; McKinney et al., 2019).

Presence of Mold

Molds may be added to the surface of dry and semi-fermented products prior to
fermentation to prevent the growth of undesirable molds and for other quality reasons,
such as flavor development. Molds are commonly added commercially by dipping or
spraying the sausage casing with a mold culture to ensure optimal mold growth/cover.
The presence of molds on the external surface may result in more uniform drying with
less chance for case hardening. Molds also may impact the drying rate particularly at
the surface, resulting in slower drying (Incze, 2010), and they may also impact the pH at
the surface of the product. Unless the drying rate/time for a product with mold is
different than the drying rate/time for a product without mold, then the presence of mold
is not considered a critical operational parameter.

Relative Humidity

It is important to maintain the relative humidity within a specified range. If the relative
humidity in the chamber is decreased too fast, rapid drying will occur, which will also
result in case hardening. When case hardening occurs, the interior moisture is
prevented from escaping, which can result in spoilage (Ruhlman and Polcyn, 2013). If
the relative humidity is too high, it can lead to undesirable mold growth. Unless there
are issues with undesirable mold growth due to high relative humidity and the product
weight loss does not meet quality/yield targets, the relative humidity does not need to be
addressed.

Air Flow

The air flow or speed (or air velocity) at which the air moves through the dryer can be
controlled by adjusting the speed at which the fan is operating. The velocity of the air
must be sufficiently high to evaporate the moisture from the surface of the food in the
dryer and sweep this moisture-rich air out of the dryer. When the heated air passes over
the moist surfaces, it picks up water through the process of evaporation. The air then
carries the water away from the food and eventually out of the dryer. As the air picks up
moisture, it cools and its moisture content increases. This reduces its ability to pick up
additional moisture as it continues its path through the rest of the dryer.

71
Air flows that are too low will not have the desired effect, and air flows too high may
increase case hardening. Unless there are issues with undesirable mold growth due to
low air flow and the product weight loss does not meet quality/yield targets, the air flow
does not need to be addressed.

72
Appendix 12: Scientific Support Available for Lethality in Dry and Semi-Dry Fermented Sausages

This guideline focuses on the safe production of fermented sausages, a category which includes such products as Lebanon bologna,
summer sausage, pepperoni, salami including Genoa, and soudjouk. Fermented sausages are generally classified as dry or semi-dry
sausages. Table 7 includes summaries of processes and the critical operational parameters of some of the processes that have been
found to achieve lethality of pathogens. Establishments are not limited to using these articles as support, and the summaries are not
adequate support on their own because they do not contain the details of each study and the establishment needs to determine if it is
representative of the actual process. For this reason, establishments will need to have the full copy of the article on-file. Links to full
copies of articles have been provided in the References section when available.

Table 7. Summary of Scientific Support Available for Lethality in Dry and Semi-Dry Fermented Sausages
Product Dia- Starter Culture Formulation Fermentation pH 6 RH 7 Low- Drying Room RH (%) Final Additional Log Reference
meter Temperature (%) Temperature Temperature Water Interven- Reduction
(mm) (°F) + Time Heat Step (°F) + Time Activity tions
Lebanon 115 Pediococcus, 3.3% salt, 2.9% Stage 1: 4.39 88 n/a n/a n/a n/a n/a >5.0-log in E. Getty, et al.,
bologna Lactobacillus, sugar, 0.8% 80°F internal ±2 coli O157:H7 1999. J Food
(beef) and dextrose, 0.14% temperature Sci. 64(6):
Micrococcus potassium for 8 hrs 1100-1107.
spp. nitrate, 0.01% (CUT 5 hrs)
potassium
nitrite Stage 2: 80
100°F hold ±
for 24 hrs 2%
(CUT 4 hrs)

Stage 3:
110°F for 24 80
hrs ±2
(CUT 2 hrs)
Smoke
applied
during last
two hours of
stage 3.

6
Target pH at the end of fermentation.
7
RH = Relative humidity (although it is a critical operational parameter it does not need to be addressed).

73
Product Dia- Starter Culture Formulation Fermentation pH 6 RH 7 Low- Drying Room RH (%) Final Additional Log Reference
meter Temperature (%) Temperature Temperature Water Interven- Reduction
(mm) (°F) + Time Heat Step (°F) + Time Activity tions
Lebanon 90 Pediococcus, 3.3% salt, 2.9% Stage 1: 4.44 88 n/a n/a n/a n/a n/a >5.0-log in E. Getty, et al.,
bologna Lactobacillus, sugar, 0.8% 80°F internal ± coli O157:H7 1999. J Food
(beef) and dextrose, 0.14% temperature 2% Sci. 64(6):
Micrococcus potassium for 8 hrs 1100-1107.
spp. nitrate, 0.01% (CUT 5 hrs)
potassium
nitrite Stage 2:
100°F hold 80
for 24 hrs ±2
(CUT 4 hrs)

Stage 3: 80
110°F for 24 ±2
hrs
(CUT 2 hrs)
Beef 105 Lactobacillus 70°F/4 days ≥5.0 1 hr at 100°F n/a n/a n/a n/a >2.0-log <5.0- Blue Ribbon
sausage plantarum followed by log in E. coli 1996
6 hrs at O157:H7
125°F
Beef 105 Pediococcus 90°F/0.5 days ≤4.6 Hold at 90°F n/a n/a n/a n/a >2.0-log <5.0- Blue Ribbon
sausage acidilactici for 7 days log in E. coli 1996
O157:H7
Beef 105 Pediococcus 90°F/0.5 days ≥5.0 Hold at 90°F n/a n/a n/a n/a >2.0-log <5.0- Blue Ribbon
sausage acidilactici for 7 days log in E. coli 1996
O157:H7
Beef 55 Pediococcus 110°F/0.7 ≥5.0 Hold at n/a n/a n/a n/a >2.0-log <5.0- Blue Ribbon
sausage acidilactici days 110°F for 7 log in E. coli 1996
days O157:H7
Beef 105 Pediococcus 110°F/0.7 ≥5.0 1 hour at n/a n/a n/a n/a >2.0-log <5.0- Blue Ribbon
sausage acidilactici days 100°F, 1 hr at log in E. coli 1996
110°F, 1 hr at O157:H7
120°F, and
ending with
7 hrs at
125°F
Beef 55 Lactobacillus 70°F/4 days ≥5.0 1 hr at 100°F n/a n/a n/a n/a >5.0-log in E. Blue Ribbon
sausage plantarum followed by coli O157:H7 1996
6 hrs at
125°F

74
Product Dia- Starter Culture Formulation Fermentation pH 6 RH 7 Low- Drying Room RH (%) Final Additional Log Reference
meter Temperature (%) Temperature Temperature Water Interven- Reduction
(mm) (°F) + Time Heat Step (°F) + Time Activity tions
Beef 55 Pediococcus 90°F/0.5 days ≤4.6 Hold at 90°F n/a n/a n/a n/a >5.0-log in E. Blue Ribbon
sausage acidilactici for 7 days coli O157:H7 1996
Beef 55 Pediococcus 90°F/0.5 days ≤4.6 1 hr at 100°F n/a n/a n/a n/a >5.0-log in E. Blue Ribbon
sausage acidilactici followed by coli O157:H7 1996
6 hrs at
125°F
Beef 105 Pediococcus 90°F/0.5 days ≤4.6 1 hour at n/a n/a n/a n/a >5.0-log in E. Blue Ribbon
sausage acidilactici 100°F, 1 hr at coli O157:H7 1996
110°F, 1 hr at
120°F, and
ending with
7 hrs at
125°F
Beef 105 Pediococcus 90°F/0.5 days ≥5.0 1 hour at n/a n/a n/a n/a >5.0-log in E. Blue Ribbon
sausage acidilactici 100°F, 1 hr at coli O157:H7 1996
110°F, 1 hr at
120°F, and
ending with
7 hrs at
125°F
Beef 55 Pediococcus 110°F/0.7 ≤4.6 Hold at n/a n/a n/a n/a >5.0-log in E. Blue Ribbon
sausage acidilactici days 110°F for 7 coli O157:H7 1996
days
Beef 55 Pediococcus 110°F/0.7 ≤4.6 Hold at n/a n/a n/a n/a >5.0-log in E. Blue Ribbon
sausage acidilactici days 110°F for 7 coli O157:H7 1996
days
Beef 105 Pediococcus 110°F/0.7 ≥5.0 Hold at n/a n/a n/a n/a >5.0-log in E. Blue Ribbon
sausage acidilactici days 110°F for 7 coli O157:H7 1996
days
Beef 66 Pediococcus 2.5% salt, 0.3% 1 hr at 86°F 5.0 80 130°F n/a n/a n/a n/a >5.0-log in E. Calicioglu, et
summer acidilactici dextrose, 0.26% 1 hr at 90°F coli O157:H7 al., 1997. J.
sausage curing salt 1 hr at 95°F Food Prot.
(6.25% sodium 1 hr at 100°F 60(1): 1158-
nitrite), 0.054% 1 hr at 105°F 1162.
sodium
erythorbate

75
Product Dia- Starter Culture Formulation Fermentation pH 6 RH 7 Low- Drying Room RH (%) Final Additional Log Reference
meter Temperature (%) Temperature Temperature Water Interven- Reduction
(mm) (°F) + Time Heat Step (°F) + Time Activity tions
Pepperoni 55 Pediococcus 3.33% salt, 96°F/14 to 18 ≤5.0 85 145°F 55°F /18 65 0.9 n/a >5.0-log in E. Hinkens, et
(beef and acidilactici 0.63% dextrose, hr internal or days coli O157:H7 al., 1996. J.
pork) 2.0% cure 128°F for 1 Food Prot.
mixture hr 59: 1260-
(156ppm 1266.
NaNO2), spice
mixture (red
pepper, clove,
anise, garlic,
oleoresin of
paprika, BHA,
BHT, citric acid).
Pepperoni 55 Pediococcus 0.63% dextrose, 96.8°F/16 to ≤4.8 92 n/a 55.4/18 days 65 0.88 Storage at >5.0-log in E. Ihnot et al.,
(beef and acidilactici 2.0% cure 20 hours 69.8 for 58 coli O157:H7 1998. J Food
pork) mixture to 60 days Micro. 40:
(156ppm 117-121. and
NaNO2), and 3% Faith et al.,
spice mixture 1997. J. of
Food
Microbiology.
37: 47-54.
Genoa 65 Pediococcus 2.9% salt, 1% 68°F/6 hrs 4.58 90- n/a 68/40 hrs 65-75 0.92 n/a >2.0-log <5.0- Porto-Fett, et
salami acidilactici, dextrose, 0.08% 80.6°F/26 hrs 95% 62.6 until log in al., 2010.
(pork) Pediococcus white pepper, 0.92 (time Salmonella Int'l J. of
pentosaceous, 0.05% sodium not and Food Micro.
and Kocuria ascorbate, reported) >1.0-log <2.0- 140: 61-75.8
varians 0.02% garlic log in E. coli
powder, 0.015% O157:H7 and
sodium nitrate, >.01-log <2.0-
and 0.005% log in Lm
sodium nitrite

76
 Product Dia- Starter Culture Formulation Fermentation pH 6 RH 7 Low- Drying Room RH (%) Final Additional Log Reference
meter Temperature (%) Temperature Temperature Water Interven- Reduction
(mm) (°F) + Time Heat Step (°F) + Time Activity tions
Genoa 105 Pediococcus 2.9% salt, 1% 68°F/6 hrs 4.58 90- n/a 68/40 hrs 65-75 0.92 n/a >2.0-log <5.0- Porto-Fett, et
salami acidilactici, dextrose, 0.08% 80.6°F/26 hrs 95 62.6 for 35 log in al., 2010.
(pork) Pediococcus white pepper, days Salmonella Int'l J. of
pentosaceous, 0.05% sodium and Food Micro.
and Kocuria ascorbate, >1.0-log <2.0- 140: 61-75.8
varians 0.02% garlic log in E. coli
powder, 0.015% O157:H7 and
sodium nitrate, >1.0-log <2.0-
and 0.005% log in Lm
sodium nitrite
Genoa 65 Pediococcus 2.9% salt, 1% 68°F/6 hrs 4.58 90- n/a 68°F /40 hrs 65-75 0.88 n/a >2.0-log <5.0- Porto-Fett, et
salami acidilactici, dextrose, 0.08% 80.6°F/26 hrs 95% 62.6 for 25 log in al., 2010.
(pork) Pediococcus white pepper, days Salmonella Int'l J. of
pentosaceous, 0.05% sodium and Food Micro.
and Kocuria ascorbate, >1.0-log <2.0- 140: 61-75. 8
varians 0.02% garlic log in E. coli
powder, 0.015% O157:H7 and
sodium nitrate, >1.0-log <2.0-
and 0.005% log in Lm
sodium nitrite
Genoa 105 Pediococcus 2.9% salt, 1% 68°F/6 hrs 4.58 90- n/a 68/40 hrs 65-75 0.94 n/a >2.0-log <5.0- Porto-Fett, et
salami acidilactici, dextrose, 0.08% 80.6°F/26 hrs 95 62.6 for 22 log in al., 2010.
(pork) Pediococcus white pepper, days Salmonella Int'l J. of
pentosaceous, 0.05% sodium and Food Micro.
and Kocuria ascorbate, >1.0-log <2.0- 140: 61-75.8
varians 0.02% garlic log in E. coli
powder, 0.015% O157:H7 and
sodium nitrate, >1.0-log <2.0-
and 0.005% log in Lm
sodium nitrite

8
Additional log reductions were achieved by use of high pressure processing (HPP) as described in the article.

77
 Product Dia- Starter Culture Formulation Fermentation pH 6 RH 7 Low- Drying Room RH (%) Final Additional Log Reference
meter Temperature (%) Temperature Temperature Water Interven- Reduction
(mm) (°F) + Time Heat Step (°F) + Time Activity tions
Soudjouk- 25mm, Bacteriocin- dextrose 75.2°F/72 4.8 90- n/a 71.6/72 80-85 0.915 Stored at >5.0-log in Porto-Fett, et
style flattened producing (0.60%), salt hours 95 hours 69.8°F Salmonella al., 2008.
semi-dry to 11.4 pediococcal (1.9%), sodium under and >2.0-log Food Micro.
sausage cm w X starter culture nitrite (0.25% or vacuum <5.0-log in E. 25: 793-801.
22.9 L 156 ppm), for 30 days coli O157:H7
(thickness chopped fresh and Lm
not garlic (0.95%),
reported) and various
spices (cumin
(0.95%), paprika
(0.42%), black
pepper (0.42%),
and all-spice
(0.42%).
Soudjouk- 3.65cm Pediococcus dextrose (1%), 71.6°F/72 4.9 50 n/a (cooking 48.2°F/18 40 not Conditione > 5.0-log in E. Calicioglu et
style natural acidilactici sodium chloride hours caused hours reported d at coli O157:H7 al., 2002. J.
semi-dry pork (2%) and 4.7% unacceptable 71.6°F/50 Food Prot.
sausage casings, spice mix (house quality) % RH for 65: 1541-
after blend, Kayseri 72 hours 1544. 9
drying Basterma, Inc., then
flattened Poughkeepsie, stored
to N.Y.). Neither under
1.25cm nitrate nor vacuum at
nitrite were 77°F for 21
used. days.

9
Additional conditions including cooking were evaluated.

78
 Product Dia- Starter Culture Formulation Fermentation pH 6 RH 7 Low- Drying Room RH (%) Final Additional Log Reference
meter Temperature (%) Temperature Temperature Water Interven- Reduction
(mm) (°F) + Time Heat Step (°F) + Time Activity tions
Soudjouk- not Pediococcus dextrose (1.5%), 75.2°F/72 4.55 90- n/a 71.6°F/72 80-85 not storage at >5.0-log in E. Calicioglu et
style reported acidilactici and sodium chloride hours 95 hours reported 77°F for 14 coli O157:H7 al., 2001. J.
semi-dry Lactobacillus (1.9%), sodium days Food Prot.
sausage curvatus nitrite (0.25% or 64: 1156-
156 ppm), 1161.9
chopped fresh
garlic (0.95%),
and various
spices (cumin
(0.95%), paprika
(0.42%), black
pepper (0.42%),
and all-spice
(0.42%).
Italian- 100 Pediococcus Nitrite, nitrate, 104°F/24 ≤4.8 >95 n/a 55.4°F/30 not ≥0.92 n/a >5.0-log IEH.
style acidilactici and spices, and salt hours days reported reduction in Evaluation of
salami Staphylococcus dextrose. Salmonella 10 Process
(pork) carnosus Parameters
NOTE: Used During
Concentrations the
were not Fermentation
reported. and Drying of
Italian-Style
Salami.
Italian- 100 Pediococcus Nitrite, nitrate, 104°F/24 ≤4.8 >95 n/a 59°F/30 days not ≥0.92 n/a >5.0-log IEH.
style acidilactici and spices, and salt hours reported reduction in Evaluation of
salami Staphylococcus dextrose. Salmonella10 Process
(pork) carnosus Parameters
NOTE: Used During
Concentrations the
were not Fermentation
reported. and Drying of
Italian-Style
Salami.

10
The amount of non-meat ingredients used was not included and none of the trials within this study were repeated. Therefore, this study should not be used alone.

79
 Product Dia- Starter Culture Formulation Fermentation pH 6 RH 7 Low- Drying Room RH (%) Final Additional Log Reference
meter Temperature (%) Temperature Temperature Water Interven- Reduction
(mm) (°F) + Time Heat Step (°F) + Time Activity tions
Salami 78 BLC-007 .57% dextrose, 88-92°F (31- < 90 n/a 67-70°F (19- 84-89 < 0.92 n/a ≥ 5.0-log10 in Deibel
0.13% white 33°C) for 28 4.9 21°C) Salmonella, Laboratories/
pepper, 0.19% hours for 24 hours E. coli CHR. Hansen.
black pepper, and then at O157:H7, and 2017.
0.014% sodium 56-59°F (13- 75-90 Lm
nitrite, and 15°C)
0.005% sodium for 29 days
nitrate
Salami 50 BLC-007 Salt, nitrite, 90°F (32°C) 4.5 95 n/a 59°F (15°C) 72 0.74 Finishing ≥ 5.0-log10 in Hussein et
spice mix for 24 hours for 31 days (after phase of Salmonella al., 2022.
drying 30 day and 4.9-log10 Food Control.
NOTE: Links were and storage at in Lm 11 131: 108432.
dipped in finishing 70°F
hydrated stage/60 (21°C) with
Penicillium days) 50% RH
nalgiovense spore
solution
Salami 80 BLC-007 Salt, nitrite, 90°F (32°C) 4.5 95 n/a 59°F (15°C) 72 0.83 Finishing ≥ 5.0-log10 in Hussein et
spice mix for 24 hours for 60 days (after phase of Salmonella al., 2022.
drying 20 day and 4.9-log10 Food Control.
NOTE: Links were and storage at in Lm11 131: 108432
dipped in finishing 70°F
hydrated stage/80 (21°C) with
Penicillium days) 50% RH
nalgiovense spore
solution

11
Although the article does not demonstrate a 5.0-log reduction for Lm, FSIS would not object to its use as supporting documentation because the article demonstrated a 4.9-log
reduction in Lm after the finishing phase.

80
Product Dia- Starter Culture Formulation Fermentation pH 6 RH 7 Low- Drying Room RH (%) Final Additional Log Reference
meter Temperature (%) Temperature Temperature Water Interven- Reduction
(mm) (°F) + Time Heat Step (°F) + Time Activity tions
Landjäger Not BLC-007 0.6% dextrose, 74°F for 72 < 61 After 71°F (21.6°C) 60 < 0.88 Raw trim ≥ 5.0-log10 in Rivera-Reyes
reported, 1.63% red wine, hours 4.8 fermentation 3 days was Salmonella, et al., 2017.
product 2.93% salt, sausages sprayed E. coli Food Control.
was 0.32% black were with 4.5% O157:H7, and 73: 767-774.
pressed pepper, 0.16 smoked with lactic acid Lm
granulated hickory (77°F/25°C
garlic, 156ppm sawdust for ) for 45s
nitrite 2 hours at and held
87°F (30°C). for 30 min.

Stored
under
vacuum
for 20 days
at 73.4F
(23°C)

81
Appendix 13. Scientific Support Available for Lethality in Salt-Cured Products
Basturma

Basturma is a traditional Turkish dry-cured beef product made from whole muscle. Basturma is known by different names depending on
geographic location: pastirma, bastirma, pasterma, basterma. To make basturma, meat is dry-cured, dried, pressed and coated with
çemen. Çemen (cement) is a paste made of spices and flavorings such as crushed cumin, fenugreek, garlic, and paprika (paste
seasoning). Table 8 includes summaries of processes and the critical operational parameters of some of the processes that have been
found to achieve lethality of pathogens. Establishments are not limited to using these articles as support, and the summaries are not
adequate support on their own because they do not contain the details of each study and the establishment needs to determine if it is
representative of the actual process. For this reason, establishments will need to have the full copy of the article on-file. Links to full
copies of articles have been provided in the References section when available.
Table 8. Summary of Scientific Support Available for Lethality in Basturma.
Product Source Formulation Dry-curing Drying Room Second drying Spice Paste and Finished Product Log Reference
mater- Temperature Step Room Third Drying Characteristics Reduction
ials (°F) + Time + Temperature Room
Relative (°F) + Time + RH Temperature (°F)
Humidity + Time + RH
(RH)
Basturma Beef 6.0 kg of curing Rounds were placed in Rounds were Rounds were Coated with a Water activity: >5.0-log in Ingham, et
rounds mixture per plastic lugs in an rinsed for 1 pressed for 12 spice paste, and 0.87 E. coli al., 2006. J
approximately alternating manner. hour with tap hours under dried 70°F for 4 O157:H7 Food Safety.
45.4 kg of Lugs were stored at water. refrigeration days MPR: 1.92: 1.00 and 26: 160-172.
meat. Curing 44°F (6.7°C), 50% RH, then dried at Salmonella
mixture for 7 days, then, fluid Dried at 70°F 70°F for 4 days. 12 hours at 65% pH: 5.6 and ≥2.0- University of
contained was drained and for 2 days RH and 12 hours log <5.0-log Wisconsin’s
proprietary rounds were manually RH cycled 12 at 80% RH % water-phase in Lm 13 Status
amounts of salt, dry-rubbed again with 12 hours at hours at 65% salt: 13.1 Summary
sucrose, a second batch of 65% RH and and 12 hours at
glucose, and curing mixture and 12 hours at 80%
sodium allowed to cure for 14 80% RH
nitrite. 12 more days.

12
Establishments wanting to use this reference should contact the authors for information on the formulation.
13
None of the trials within this study were repeated. So, establishments should provide additional supporting documentation for the processes effectiveness.

82
 Product Source Formulation Dry-curing Drying Room Second drying Spice Paste and Finished Product Log Reference
mater- Temperature Step Room Third Drying Characteristics Reduction
ials (°F) + Time + Temperature Room
Relative (°F) + Time + RH Temperature (°F)
Humidity + Time + RH
(RH)
Basturma Beef 6.0 kg of curing Rounds were placed in Rounds were Rounds were Coated with a Water activity: >5.0-log in Ingham, et
rounds mixture per plastic lugs in an rinsed for 1 pressed for 12 spice paste, and 0.95 E. coli al., 2006. J
approximately alternating manner. hour with tap hours under dried 75°F for 5 O157:H7 Food Safety.
45.4 kg of Lugs were stored at water. refrigeration days MPR: 2.00: 1.00 and 26: 160-172.
meat. Curing 44°F (6.7°C), 50% RH, then dried at Salmonella
mixture for 7 days, then, fluid Dried at 75°F 75°F for 4 days 12 hours at 65% pH: 6.0 and ≥2.0- University of
contained was drained and for 2 days RH and 12 hours log <5.0-log Wisconsin’s
proprietary rounds were manually Cycled 12 hours at 80% RH % water-phase in Lm1 Status
amounts of salt, dry-rubbed again with 12 hours at at 65% RH and salt: 8.3 Summary
sucrose, a second batch of 65% RH and 12 hours at 80%
glucose, and curing mixture and 12 hours at RH
sodium allowed to cure for 14 80% RH
nitrite. 14 more days.
Basturma Beef 6.0 kg of curing Rounds were placed in Rounds were Rounds were Coated with a Water activity: >5.0-log in Ingham, et
rounds mixture per plastic lugs in an rinsed for 1 pressed for 12 spice paste, and 0.84 E. coli al., 2006. J
approximately alternating manner. hour with tap hours under dried 81°F for 6 O157:H7 Food Safety.
45.4 kg of Lugs were stored at water. refrigeration days MPR: 1.52: 1.00 and 26: 160-172.
meat. Curing 44°F (6.7°C) , 50% RH, then dried at Constant 70% Salmonella
mixture for 7 days, then, fluid Dried at 81°F 81°F for 4 days pH: 5.6 and ≥2.0- University of
contained was drained and for 2 days log <5.0-log Wisconsin’s
proprietary rounds were manually Constant 70% % water-phase in Lm1 Status
amounts of salt, dry-rubbed again with Constant 70% salt: 18 Summary
sucrose, a second batch of
glucose, and curing mixture and
sodium allowed to cure for 14
nitrite. 15 more days.

14
Establishments wanting to use this reference should contact the authors for information on the formulation.
15
Establishments wanting to use this reference should contact the authors for information on the formulation.

83
Product Source Formulation Dry-curing Drying Room Second drying Spice Paste and Finished Product Log Reference
mater- Temperature Step Room Third Drying Characteristics Reduction
ials (°F) + Time + Temperature Room
Relative (°F) + Time + RH Temperature (°F)
Humidity + Time + RH
(RH)
Basturma Beef Injected with 40°F for up to 6 days Muscles hung 68-77°F for 4 Mixture of spices ≥5.0-log in Genigeorgis
eye 10% with frequent individually in more days (flour, fenugreek, Salmonella and
round (weight/weight) rotation. smokehouse pepper, cumin, Lindroth.
whole brine made of and heated at garlic, paprika, 1984.
or cut 25% salt and an oven and food color) in European
longitu 0.2% sodium temperature water. Meeting of
dinally nitrite. After of ≥ 149°F for Dried at 68-77°F Meat
in half injection dry at least 6 for 4 more days Research
salt added at hours during Workers.
levels of 4.7- which 217-224.
10% (w/w) internal meat
temperature
reached ≥
127 °F.

84
Country Cured Ham

Country cured ham is the dry cured hind leg of a pig. FSIS has a standard of identity for country ham in 9 CFR 319.106. The
production process for country cured hams is similar to prosciutto and involves the three steps of curing, equalization, and drying.
Table 9 contains a summary of processes and the critical operational parameters of some of the processes that have been found to
achieve lethality of pathogens. Establishments are not limited to using these articles as support, and the summaries are not adequate
support on their own because they do not contain the details of each study and the establishment needs to determine if it is
representative of the actual process. For this reason, establishments will need to have the full copy of the article on-file. Links to full
copies of articles have been provided in the References section when available.

Table 9. Summary of Scientific Support Available for Lethality in Country Cured Ham.
Product Formulation Cure Dry-curing Equalization Drying Log Reduction Finished Product Reference
application Characteristics
Country cured 1) 3.63 kg salt, Cures were Hams were Hams were Hams were ≥5.0-log in E. coli Finished product day Reynolds, et al.,
ham 454 g sugar, applied at a cured at 40°F placed in dry aged for O157:H7 and = 120 2001. Journal of
14.2 g sodium ratio of 42.53 (4.4°C) during stockettes and 20 d at 85°F Salmonella and ≥4-log Salt = 8.0% Food Science.
nitrite, 56.7 g g of cure dry salt curing held at 40°F (29.4°C) (65% in Lm when cured for water activity = 0.91 66: 1373-1379.
sodium nitrate mixture per (35 d), after (4.4°C) for 14 relative 69 days. pH = 5.5
0.45 kg of which the d for salt humidity) to Brine concentration =
OR ham, with a excess salt equalization. meet Trichinae Research also validated 13.81% and 12.33%
half portion was brushed requirements process resulted in for cure mix 1 and
2) 3.63 kg salt each of this off with no in 9 CFR sufficient control of S. cure mix 2,
and 454 g cure mixture water added. 318.10. 16 aureus growth. respectively.
sugar applied in 2
separate hand
rubs, on day 0
and day 10.

16
Dry curing process was considered complete at this point (d 69). At d 69, hams were placed in ambient 68 to 75.2°F (20 to 24 °C) storage through d 120. Sufficient reduction of
pathogens was achieved after d 69.

85
Bresaola

Bresaola is traditionally produced with whole beef muscles, such as eye of round and inside round. These cuts are typically dry-cured
with salt and spices, allowed to equalize, and then stuffed into casings and hung to dry (Watson et al., 2021). Table 10 contains a
summary of processes and the critical operational parameters of some of the processes that have been found to achieve lethality of
pathogens. Establishments are not limited to using these articles as support, and the summaries are not adequate support on their own
because they do not contain the details of each study and the establishment needs to determine if it is representative of the actual
process. For this reason, establishments will need to have the full copy of the article on-file. Links to full copies of articles have been
provided in the References section when available.

Table 10. Summary of Scientific Support Available for Lethality in Bresaola.

Product Formulation Cure Dry-curing Equalization Drying Log Reduction Finished Product Reference
application Characteristics
Bresaola Whole muscle The eye of After being The eye of Bresaola was ≥5.0-log in E. coli Water activity = 0.88 Watson, et al.,
intact beef round coated with round stuffed into O157:H7, Salmonella 2021. Meat and
rounds were subprimals dry remained at beef bung and Lm when cured for Muscle Biology.
sprayed with were cured by ingredients, 39°F (4°C) for casings (115- 65 days. 5(1): 14, 1-8.
Beefxide (2.5% applying half beef was 4 weeks. 130mm) and
vol/vol) using of the cure placed in a were dry aged Results were calculated
a handheld mixture before meat lug at at 54 to 57°F on the surface and
sprayer for 15- dry-curing, 39°F (4°C) for (12-14°C) (65- therefore, should only
20 sec on each after 7 days 7 days before 75% relative be applied when using
side to coat all the subprimals being coated humidity) for whole muscle intact
surfaces. were coated with the 65 days. beef source materials.
A proprietary with the remaining half
blend of salt remaining half of the cure
(3.5% of meat of cure mixture.
weight) and mixture.
cure
ingredients
(150ppm
sodium nitrite
and 100 ppm
sodium
nitrate).

86
Appendix 14: Scientific Support Available for Lethality in Dried Products

Droëwors

Droëwors is a dried beef sausage product developed in South Africa that can be made from various meat species (typically beef or
game). Droëwors is made from ground meat stuffed into casings after being mixed with vinegar and seasoning blends. Table 11
includes a summary of the processes and the critical operational parameters of some of the processes that have been found to achieve
lethality of pathogens. Establishments are not limited to using these articles as support, and the summaries are not adequate support
on their own because they do not contain the details of each study and the establishment needs to determine if it is representative of
the actual process. For this reason, establishments will need to have the full copy of the article on-file. Links to full copies of articles
have been provided in the References section when available.

Table 11. Summary of Scientific Support Available for Lethality in Droëwers.

Product Product Formulation Drying Post-drying Finished Product Log Reduction Reference
Characteristics Storage Characteristics
Droëwers Grind size: Vinegar/spices: Droewors sausage After Trial 1 ≥2.0-log and < 5.0- Burnham, et al.,
Trimmings Ground meat was hung on racks in reaching a pH: 5.5 log reduction in E. 2008. Journal of
were ground was stuffed into an environmental water water activity: 0.62 coli O157:H7 and Food Safety. 28:
twice, first vacuum packed chamber set to 72°F activity of % water-phase salt: 19.1% Salmonella and a 1.5 198-209.
through a plate bags and (actual range 68- 0.60, to 2.7-log reduction
with 10 mm proprietary 71.6°F) with a target sausages Trial 2 in Lm.
size holes and amounts of constant of 50% were pH: 5.4
then through a vinegar and a relative humidity vacuum water activity: 0.60 Research also
plate with 4mm spice blend (actual range 38- packaged % water-phase salt: 19.6% validated process
size holes. were added. 64%). and stored resulted in sufficient
Then the mix Sausages were dried at 68-71.6°F Trial 3 control of S. aureus
was stuffed into for 12-21 days. for 7 days. pH: 5.4 growth.
lamb casings. 17 water activity: 0.60
% water-phase salt: 22.2%

17
Establishments wanting to use this reference should contact the authors for information on their vinegar and spice formulation or should ensure that the pH and % water phase-
salt reported match those listed in the “finished product characteristics” column.

87
Biltong

Biltong is a dried beef product developed in South Africa. Traditionally, biltong is dried under ambient conditions; however, to achieve adequate
lethality commercially produced biltong should be produced under controlled conditions. Biltong may be produced from various species (e.g., beef,
poultry, venison, or game). In the U.S. biltong is most often made from strips of beef that are trimmed, salted, and dried. Consumption of biltong
has been linked to at least one outbreak from Salmonella outside of the U.S. and in a 2008 outbreak, one case patient was an infant who was given
the product for teething (Mindlin et. al, 2013). The intended use of biltong is typically considered to be RTE because consumers commonly consume
biltong without further preparation for safety. Table 12 includes summaries of processes and the critical operational parameters of some of the
processes that have been found to achieve lethality of pathogens. Establishments are not limited to using these articles as support, and the
summaries are not adequate support on their own because they do not contain the details of each study and the establishment needs to determine
if it is representative of the actual process. For this reason, establishments will need to have the full copy of the article on-file. Links to full copies of
articles have been provided in the References section when available.

Table 12. Summary of Scientific Support Available for Lethality in Biltong.

Product Product Formulation Drying Post- Finished Product Log Reduction Reference
Characteristics drying Characteristics
Storage
Biltong Thickness: Biltong Vinegar/spices: Biltong strips After Trial 1 ≥ 2.0-log Burnham, et
strips were Proprietary were hung on reaching a pH: 5.6 reduction and < al., 2008.
obtained from amounts of vinegar racks in an water water activity: 0.75 5.0-log reduction Journal of
intact beef bottom and spice blend. environmental activity of % water-phase salt: 15.4% for Salmonella Food Safety.
round sub-primal Tumbled for 30 chamber set to 0.60, and E. coli 28: 198-209.
cuts which were minutes. 18 72°F (actual strips Trial 2 O157:H7 and ≥
trimmed of range 68-71.6°F) were pH: 5.6 1.0-log reduction
external flat, with a target of vacuum water activity: 0.75 and ≤ 4.0-log
squared off to 50% relative packaged % water-phase salt: 15.8% reduction for Lm.
produce a uniform humidity (actual and stored Also validated the
shape, and sliced range 38-64%). at 68- Trial 3 process resulted
to 2.5 cm Strips were 71.6°F for pH: 5.5 in sufficient
thickness. dried for 17-26 7 days. water activity: 0.62 control of S.
days. % water-phase salt: 21.5% aureus growth.

18
Establishments wanting to use this reference should contact the authors for information on their vinegar and spice formulation or should ensure that the pH and % water phase-
salt reported match the actual process.

88
 Product Product Formulation Drying Post- Finished Product Log Reduction Reference
Characteristics drying Characteristics
Storage
Biltong Beef steak portions Marinated in Meat pieces n/a Not reported. ≥ 5.0-log Naidoo, et
(tradit- (30cm length x undiluted apple were dried in a reduction for al., 2010.
ional 15cm width x cider vinegar (100 Biltong buddy Lm. 20 Food
method) 2.5cm thickness). ml) for 30 seconds home-dryer Control. 21:
per side. Excessive fitted with a The research also 1042-1050.
vinegar was allowed 40W light bulb validated the
to drip off and 16 that generated a process resulted
grams biltong spice constant in sufficient
(predominantly temperature of control of S.
consisting of black 77°F. To aureus growth.
pepper, coriander, achieve
salt and brown adequate
sugar) was spread reductions
onto each side of pieces were
the meat. Pieces dried for 84
were then hours. Air flow
marinated for 20 and other
hours under critical
refrigeration (40°F). operational
parameters of
the dryer were
not reported. 19

19
Critical operational parameters used in the study including the temperature were not reported. Therefore, establishments should provide additional supporting documentation for
the effectiveness.
20
Establishments should provide additional supporting documentation for the effectiveness because only one strain of Lm was used and no history or justification was given for why
the particular Lm strain was chosen.

89
 Product Product Formulation Drying Post- Finished Product Log Reduction Reference
Characteristics drying Characteristics
Storage
Biltong Beef steak portions 16 grams biltong Meat pieces n/a Not reported. ≥ 5.0-log Naidoo, et
(modern (30cm length x spice were dried in a reduction for al., 2010.
method) 15cm width x (predominantly Biltong buddy Lm. 22 Food
2.5cm thickness) consisting of black home-dryer Control. 21:
pepper, coriander, fitted with a The research also 1042-1050.
salt and brown 40W light bulb validated the
sugar) was that generated a process resulted
combined with constant in sufficient
16ml apple cider temperature of control of S.
vinegar (1:1 g/ml 77°F. To aureus growth.
ratio) and was achieve
spread onto each adequate
side of the meat reductions
and sealed and pieces were
shaken for 1 minute dried for 96
to ensure full hours. Air flow
coverage. Pieces and other
were then critical
marinated for 20 operational
hours under parameters of
refrigeration (40°F). the dryer were
not reported. 21

21
Critical operational parameters used in the study including temperature were not reported. Therefore, establishments should provide additional supporting documentation for the
effectiveness.
22
Establishments should provide additional supporting documentation for the effectiveness because only one strain of Lm was used and no history or justification was given for why
the particular Lm strain was chosen.

90
 Product Product Formulation Drying Post- Finished Product Log Reduction Reference
Characteristics drying Characteristics
Storage
Biltong Boneless beef Raw beef strips 77°F (25°C) and n/a 0.90 water activity 23 ≥ 5.0-log Karolenko, et
round was were dipped in 55% RH for at reduction for al., 2020.
trimmed to remove acidified calcium least 4 days. Salmonella Microorganis
excess fat and sulfate (Mionix RTE- ms. 8(5):
trimmed to 1.9 cm 01) adjusted to 10% 791.
thick X 5.1 cm wide lactic acid for 30
X 7.6cm long. seconds.

Strips were then

tumble-marinaded
for 30 minutes
under vacuum in a
marinade
containing salt (2%
of formulation) and
vinegar (2%
formulation).

23
Results for each trial were not reported. Water activity estimated based on one trial reported in Figure 7. Product may need to be dried further for shelf-stability.

91
 Product Product Formulation Drying Post- Finished Product Log Reduction Reference
Characteristics drying Characteristics
Storage
Biltong Boneless beef Raw beef strips 77°F (25°C) and 0.85 water activity 24 ≥ 5.0-log Karolenko, et
round was were dipped in 55% RH for at reduction for al., 2020.
trimmed to remove acidified calcium least 6 days. Salmonella Microorganis
excess fat and sulfate (Mionix RTE- ms. 8(5):
trimmed to 1.9 cm 17) adjusted to 5% 791.
thick X 5.1 cm wide lactic acid for 30 or
X 7.6cm long. 60 seconds or 5%
lactic acid for 60
seconds.

Strips were then

tumble-marinaded
for 30 minutes
under vacuum in a
marinade
containing salt (2%
of formulation) and
vinegar (2%
formulation).

24
Results for each trial were not reported. Water activity estimated based on one trial reported in Figure 7.

92
 Product Product Formulation Drying Post- Finished Product Log Reduction Reference
Characteristics drying Characteristics
Storage
Biltong Boneless beef Raw beef strips 77°F (25°C) and 0.90 water activity 25 ≥ 5.0-log Karolenko, et
round was were dipped in 5% 55% RH for at reduction for al., 2020.
trimmed to remove lactic acid or 3% least 4 days Salmonella Microorganis
excess fat and sodium acid sulfate ms. 8(5):
trimmed to 1.9 cm for 30 seconds. 791.
thick X 5.1 cm wide
X 7.6cm long. Strips were then
tumble-marinaded
for 40 minutes
under vacuum in a
marinade
containing salt (2%
of formulation) and
vinegar (2%
formulation).

Marinaded beef
was held overnight
(16-18hours) at
41°F (5°C).

25
Results for each trial were not reported. Water activity estimated based on one trial reported in Figure 7. Product may need to be dried further for shelf-stability.

93
 Product Product Formulation Drying Post- Finished Product Log Reduction Reference
Characteristics drying Characteristics
Storage
Biltong Boneless beef Raw beef strips 73°F (22.8°C) 0.85 water activity 26 ≥ 5.0-log Karolenko, et
round was were dipped in and 55% RH for reduction for al., 2020.
trimmed to remove water for 30 at least 6 days Salmonella Microorganis
excess fat and seconds. ms. 8(5):
trimmed to 1.9 cm 791.
thick X 5.1 cm wide Beef strips were
X 7.6cm long. tumbled (no
vacuum) for 5 min
with spices (95-96%
beef, 4-5% spice
which included salt
at 2.1% total
formulation). Dry
spiced-beef strips
were placed in
stainless steel pans
and liquid marinade
was slowly poured
over (vinegar
comprised 73%
liquid marinade and
10% total
formulated weight).
Marinaded pieces
were held at 41°F
(5°C) and turned
after 30 min and 8-
12 hours (total time
16-20 hours).

26
Results for each trial were not reported. Water activity estimated based on one trial reported in Figure 7.

94
Appendix 15: Designing Challenge Studies for Fermented, Salt-Cured,
and Dried Products

If no literature is available or the process or method an establishment uses is

significantly different than used in literature, establishments may decide to conduct a
challenge study. When establishments want to use unique processes, a challenge
study may be needed to demonstrate the safety of the process. Before conducting a
challenge study, establishments may consider reaching out to the HACCP Coordinator
in their state who can provide technical advice, assistance, and resources to support
HACCP implementation in small and very small plants.

For general guidance on conducting challenge studies see the FSIS HACCP Systems
Validation Guideline as well the article, Parameters for Determining Inoculated
Pack/Challenge Study Protocols, published by the National Advisory Committee on
Microbiological Criteria for Foods (NACMCF) in the Journal of Food Protection in 2010.
For guidance on how to select a microbiological testing laboratory please review FSIS’
Establishment Guidance for the Selection of a Commercial or Private Microbiological
Testing Laboratory. Specific considerations related to fermented, salt-cured, and dried
meat and poultry products that are not addressed in other guidance are included below.

A well-designed and documented challenge study should include details about:

• Product studied including formulation information.
• The types and number of strains of microorganisms.
• Methods of production, enumeration, and standardization of inoculum.
• Size of the inoculum to be used.
• Method of inoculation.
• Sample size, sampling time, and number of samples to test.
• Methods of microbial analysis.
• Number of replicates.
• Measurement of critical operational parameters and intrinsic factors at each key
stage of production.

Product Studied Including Formulation Information

The study should clearly identify the type of product studied as well as details about the
raw meat or poultry components used (e.g., biltong strips were obtained from intact beef
bottom round sub-primal cuts which were trimmed of external flat, squared off to
produce a uniform shape, and sliced to 2.5 cm thickness).

The challenge study should identify all ingredients used in the product formulation which
may affect the inactivation or growth of bacterial pathogens in or on the product to
include ingoing levels of sodium nitrite, phosphate, preservatives, and any antimicrobial
agents (e.g., organic acid salts).

95
The Types and Number of Strains of Microorganisms

The types and number of strains of microorganisms used as an inoculum should be

included in the protocol as well as an explanation of why the strains (including
surrogates) were chosen.

If an establishment chooses to conduct a challenge study in a laboratory, the study

should use at least five strains of the pathogenic microorganisms of concern (e.g., five
strains of Salmonella, five of STEC, and five of Lm) including strains associated with
human foodborne illness and strains isolated from meat and poultry products.

If an establishment chooses to conduct a challenge study in a plant environment to

best represent actual processing conditions, the establishment should choose non-
pathogenic surrogate organisms that have been found to respond similarly to the
pathogens of interest (e.g., Salmonella, S. aureus, etc.) under the mode of lethality
being evaluated (e.g., fermentation, salt-curing, or drying). See Table 13 for examples.

FSIS recommends establishments target the same reductions in surrogates as they

would for pathogens (e.g., a 5-log reduction for surrogates found to respond similarly to
Salmonella and STEC and a 3-log reduction for surrogates found to respond similarly to
Lm). It may be possible for establishments to use adjustment factors to account for
differences in tolerance to lethality treatments between a surrogate and pathogen.
However, using adjustment factors is not recommended by FSIS unless the
establishment also has performed a challenge study in the laboratory to support the
effectiveness of the lethality treatment in reducing pathogens under laboratory
conditions.

An establishment that chooses to do a challenge study may use a surrogate organism

to measure change, but it should do so only after giving careful consideration to specific
precautions. These precautions include ensuring that a microbiologist trained in food
science and in the design of inoculated-pack studies introduces the non-pathogenic
cultures within the establishment. In addition, the establishment should ensure that the
introduction of the non-pathogenic cultures does not create an insanitary condition in
the facility or cause the food to become adulterated. Whenever possible, the
establishment should also avoid the use of bacterial surrogate strains that have been
'marked' using antibiotic resistant genes. Finally, establishments should ensure that the
non-pathogenic cultures are necessary and proven to be effective for the intended
purpose.

To better ensure that insanitary conditions are not created, establishments are
encouraged to apply surrogate indicator organism cultures in a manner to ensure that
the establishment can conduct a full cleaning and sanitizing of the facility and
equipment after the stage in the food safety application being evaluated. Generally,
product containing the surrogate indicator organism cultures would not automatically be
considered adulterated.

Additional information on the strengths and limitations for using a particular surrogate
indicator organism is provided in scientific research and should be considered.

96
 Table 13. Potential surrogates for lethality challenge studies conducted in-plant.

If you produce… the following to respond for the as supported by… 27

surrogate(s) have similarly following
been validated… to… processing
step(s)…
Summer sausage Five American Type Salmonella, Fermentation Kneeling et al., 2009
and biltong Culture Collection E. coli 28 Niebuhr et al., 2008
(ATCC) non- O157:H7 Karolenko et al., 2022
pathogenic E. coli: https://askusdacontactcente
• BAA-1427 For biltong, r.force.com/s/article/Use-
• BAA1428 also Lm and of-Non-pathogenic-
• BAA-1429 S.aureus Escherichia-coli-E-coli-
• BAA-1430 Cultures-as-Surrogate-
• BAA-1431 Indicator-Organisms-in-
Validation-Studies
Fermented and Enterococcus Salmonella Fermentation, Ma et al., 2007
dried salami, Dry- faecium drying, and Michet, 2015
cured whole dry-curing 29 Guidelines for Using
muscle pork Enterococcus faecium as a
products See NOTE Surrogate Microorganism in
below Almond Process Validation
Whole-muscle and Saga 200 (Lactic Salmonella, Antimicrobial Borowski et al., 2009
ground and formed acid bacteria – E. coli treatment and Dierschke et al., 2010a
jerky, Pediococcus O157:H7, drying Dierschke et al., 2010b
Biltong, acidilactici) Lm, and See NOTE
Droëwors S. aureus below
Biltong Carnobacterium Salmonella, Antimicrobial Karolenko et al., 2022
spp. E. coli treatment and
O157:H7, Drying
Lm, and
S. aureus

NOTE: Research by Karolenko et al., 2022 showed Enterococcus faecium and Saga
200 to be much more tolerant to drying during biltong processing compared to
Salmonella and other pathogens. Other surrogates such as Carnobacterium spp. and
the Five American Type Culture Collection (ATCC) non-pathogenic E. coli were found to
behave more similarly to Salmonella and other pathogens. For this reason, FSIS
recommends establishments consider other surrogates such as Carnobacterium spp. or
the Five American Type Culture Collection (ATCC) non-pathogenic E. coli for in-plant
validation studies used to validate drying.

27
Additional research may be available to support the use of these (or other) surrogates for these (or
other) processing steps.
28
The surrogates have also been validated for freezing, refrigerated storage, and antimicrobial treatment
of carcasses and ground beef.
29
The surrogate has also been validated for cooking of ground beef.

97
Methods of Production, Enumeration, and Standardization of Inoculum

FSIS recommends that strains used in challenge studies for processes that rely on low
pH to achieve lethality (e.g., fermentation) be exposed to acid during inoculum
preparation (Breidt et al., 2018; Buchanan and Edelson, 1996; Hill et al., 1995). Adding
glucose is one way to ensure that the inoculum is pre-adapted for acid tolerance,
because the cells ferment the glucose, thereby lowering the pH of the media.
Specifically, for studies conducted in a testing laboratory using STEC and Salmonella,
individual cultures of each strain should be prepared by inoculating an appropriate
growth media, such as tryptic soy or trypticase soy broth, supplemented with 1%
glucose and incubating for 18 to 24 hours at 98.6 °F (37 °C) without agitation (shaking)
to obtain stationary phase cells. Cultures should be grown the day prior to product
inoculation with a minimum holding period prior to actual use. Each strain should be
centrifuged, washed, and resuspended in 0.1% peptone broth. Dilutions of each strain
should be made to yield approximately equal numbers of each of the five strains. The
five strains should be thoroughly mixed prior to being used as an inoculum. After the
mixed working inoculum is prepared, the viable count of the mixture should be
determined by direct surface plating on sorbitol-MacConkey agar. Each of the individual
strains in the inoculum should contribute about 20% of the total inoculum (Eblen et al.,
2005).

Size of the Inoculum to be Used

The final concentration of Salmonella, STEC, or Lm in the product mixture should be no

less than 2.0 X 107 CFU/g of meat mixture. The actual inoculum level in the product
mixture should be confirmed by sampling the inoculated meat mixture immediately after
inoculation. At this concentration, product can be serially diluted and direct plated
without the need for enrichment to recover low levels of inoculum. The initial inoculum
level was chosen to allow direct enumeration of at least a 5.0-log reduction in the level
of the inoculum between the initial count in the product mixture and the finished product.

Method of Inoculation

The inoculum should be added to the meat (or poultry) mixture prior to the addition of
the other ingredients or a starter culture.

The following procedure is recommended for fermented sausages:

• Add inoculum to meat or poultry while grinding or chopping to desired

consistency.
• Mix in cure (if used), salt, and spices.
• Blend in starter culture near end of mixing cycle.
• Stuff batter into casings.

NOTE: For practical reasons, establishments may formulate their products at the
establishment and send the batter to their private lab where the inoculum is mixed in
prior to stuffing. FSIS has no objection to this procedure provided all replicates are
prepared in the same manner.

98
For fermented products, inoculated product should be stuffed into casings as usual to
approximate normal production procedures. A shorter length in casings may be used as
long as the length is approximately twice the diameter of the stuffed casing.

For fermented sausages, beaker sausages (where the sausage batter is fermented in
a test tube) may be used to give a general estimate of the effectiveness of a process
but are not recommended as the basis of a model system in a study used as the sole
support because of differences in geometry and size of the test tube compared to the
product produced and because of differences in the processing conditions, including the
humidity. See pages 13 of this document for an explanation of how use of impermeable,
sealed glass tubes as a model system may have contributed to an outbreak in a
Lebanon bologna product.

Sample Size, Sampling Time, and Number of Samples to Test

A minimum of two samples for microbiological analysis should be collected at Time 0

and at the end of all multi-hurdle steps. Time 0 should be the point after the inoculum is
added to the product but before the processing steps are applied. Time 0 should not be
the inoculum level in the test tube before it is applied to the product.

For dry or semi-dry fermented sausage, establishments should select two sausage
sticks at the end of drying (finished product). From each stick selected, cut multiple
cross-sectional slices from multiple locations on each stick to a final analytical sample
weight based on the method (e.g., many methods are validated for 25 g).

Methods of Microbial Analysis

Methods should be specific or fit for the intended purpose to detect the target
microorganism in the sample.

Number of Replicates

As recommended by NACMCF, at least two replicates should be conducted when three

or more samples are tested at each time interval. According to NACMCF, replicates
should be independent trials using different lots of product and inoculum to account for
variations in product, inoculum, and other factors. When the number of samples
analyzed at each time interval is only two, it is better for the study to be repeated
(replicated) more than two times.

For guidance on evaluating the results of a challenge study or other scientific literature
see FSIS’ Cooking Guideline, page 56, “Acceptability of Challenge Study Results”.

Measurement of Critical Operational Parameters and Intrinsic Factors at Each Key

Stage of Production

Although samples for microbiological analysis are recommended to be collected at Time

0 and at the end of all of the multi-hurdle steps (e.g., fermentation and drying or salt-
curing and drying), FSIS recommends data be gathered on the critical operational
parameters (e.g., time, temperature, and relative humidity) during each processing step
(e.g., stuffing, fermentation, low-temperature heat step, if used, and drying). In addition,
99
data on the product’s intrinsic factors (e.g., salt and brine concentration, pH, and water
activity) should be gathered at the end of each key processing step (e.g., after stuffing,
fermentation, low-temperature heat step, if used, and after drying). Parameters
assayed for at the end of each step may also include moisture, fat, protein, and
titratable acidity.

The challenge study should be designed to closely match the critical operational
parameters (e.g., time, temperature, and relative humidity) and product’s intrinsic
factors (e.g., salt and brine concentration, pH, and water activity) in the establishment’s
actual process. As with journal articles and other types of support, it is very important
that the critical operational parameters used during the actual experiment are
documented, as these should then set the range for the establishment’s actual process
critical control point (CCP) critical limits and program limits. For example, if a challenge
study is conducted to measure the effectiveness of a drying step and the product is
dried in a drying room where the temperature ranged from 50-60 °F, the relative
humidity ranged from 45-55%, and the air flow ranged from 0.5 to 1 m/sec, then the
establishment should set its CCP limits to ensure the temperature does not go below 50
°F or above 60 °F, the relative humidity should not go below 45% or above 55%, and
the air flow does not go below 0.5 m/sec or above 1 m/sec. If the establishment wants
to use different limits, then it should provide a science-based justification for why these
different limits would result in reductions consistent with those observed in the challenge
study.

100
Appendix 16: Glossary

Air velocity: rate of motion of air in a given direction. 30

Alternative lethality: A lethality target or log reduction that is different from FSIS
recommendations but achieves an equivalent probability that no viable Salmonella
organisms or other pathogens of concern remain in the finished product when properly
implemented as described in the supporting scientific documentation.

Bacteriocins: toxins produced by bacteria that inhibit the growth of other similar
bacteria.

Basturma: a traditional Turkish product made from whole muscle that is dry-cured,
dried, pressed and coated with çemen, a paste made of spices and flavorings, such as
crushed cumin, fenugreek, garlic, and paprika. Other names for the product include
pastirma, bastirma, pasterma, basterma.

Biltong: a dried beef product developed in South Africa that is often made from strips of
beef that are trimmed, salted, and dried.

Bresaola: is a salt-cured and dried product traditionally produced with whole beef
muscles, such as eye of round and inside round. These cuts are typically dry-cured
with salt and spices, allowed to equalize, and then are stuffed into casings and hung to
dry (Watson et al., 2021).

Brine Concentration: is a measure of the amount of salt in the water phase of the
product. Brine concentration can’t be determined by the formulation; it is a value
calculated from the total salt content and total water content values obtained by a lab
analysis. Refer to FSIS Processing Inspectors’ Calculations Handbook Chapter 14 for
more information.

Case hardening: a shell of hardened protein at the surface of a meat or poultry

product.

Casing type: the type of material that encloses the filling of a sausage.

Certificates of Analysis (COAs): A document that includes test results or other

analyses to show a product conforms to its specifications.

Challenge study: a type of study used to simulate what happens to a product during
processing, distribution, and subsequent preparation and handling should it become
contaminated 31.

Country ham: the dry-cured hind leg of a pig that is produced meeting the standard of
identity in 9 CFR 319.106.

30
https://www.mindat.org/glossary/air_velocity
31
https://www.foodsafetymagazine.com/magazine-archive1/aprilmay-2001/do-you-need-microbial-challenge-
testing/
101
Critical operational parameters: the specific conditions that the intervention must
operate under for it to be effective.

Degree-hours: the amount of time in hours above 60 °F (the critical temperature at

which staphylococcal growth effectively begins) an establishment’s fermentation
process can take at a specific temperature to reduce the pH to 5.3 or below to control S.
aureus growth. Degree-hours were specifically designed to control S. aureus growth
during fermentation.

Direct acidulation: the process of reducing the pH by the direct addition of organic
acids.

Dried products: comminuted, sliced whole muscle, or whole muscle products that may
be formulated with nitrite, may be smoked, are usually heated, and are dried.

Droëwors: a dried beef sausage product developed in South Africa that is made from
ground product stuffed into casings after being mixed with vinegar and seasoning
blends.

Dry-curing: process where the cure ingredients are rubbed onto the food surface or
mixed into foods.

Excision sampling: sampling method that involves cutting samples of meat from the
surface.

Fermented products: products in which the raw meat or poultry component is usually
reduced in size, formulated with cure, starter culture, salt and seasoning mixture, stuffed
in casings, fermented, sometimes heated with a low temperature heat step for food
safety or smoked, and then dried.

Genoa salami: A type of dry sausage product prepared with all pork or with a mixture of
pork and a small amount of beef. For more information see the entry in FSIS Food
Standards and Labeling Policy Book available at:
https://www.fsis.usda.gov/guidelines/2005-0003.

Green weight: the weight of the meat and/or poultry product or meat and/or poultry
byproduct (meat block) before processing.

Isoelectric point: the pH where positive charges are equal to negative charges. For
meat or poultry, once the pH has reached the isoelectric point of major proteins, the
positive charges are equal to negative charges resulting in a net charge of the protein of
zero. These positive and negative groups within the protein are attracted to each other
and can result in a reduction in the amount of water than can be attracted and held by
that protein. 32

Lebanon bologna: A type of coarse ground, fermented, semi-dry sausage. The

production of Lebanon bologna typically involves combining curing ingredients (such as

32
https://swine.extension.org/water-holding-capacity-of-fresh-meat/
102
salt and sodium nitrite) and a "starter" culture of lactic acid bacteria with coarse ground
beef. This mixture is placed in casings, fermented, and further dried.

Lethality: the process or combination of processes that ensures that no Salmonella

organisms remain in the finished product, as well as reduces other pathogens and their
toxins or toxin metabolites. Examples of lethality processes include cooking,
fermentation, salt-curing, and drying.

Letters of Guarantee (LOGs): A LOG is a document that provides details for

components that are used in the areas of food processing, handling, and storage.

Moisture Protein Ratio (MPR): a measure that expresses the percent moisture divided
by the percent protein. MPR is commonly used in the U.S. to classify dried sausages
and other meat products. Although MPR values indicate the degree of product drying,
they are not necessarily indicative of microbial safety or product shelf-stability because
they do not take into account availability of the water.

Mother batch: previously fermented and controlled batch of product.

Pepperoni: A type of dry sausage prepared from pork or pork and beef. For more
information see the entry in available at: https://www.fsis.usda.gov/guidelines/2005-
0003.

Proteolysis: the breakdown of proteins or peptides into amino acids by the action of
enzymes.

Ready-to-eat meat and poultry product is defined by FSIS in 9 CFR 430.1 as a meat or
poultry product that is in a form that is edible without additional preparation to achieve
food safety and may receive additional preparation for palatability or aesthetic,
epicurean, gastronomic, or culinary purposes.

Salt-cured products: usually whole muscle products that are cured with salt and
sodium nitrite and/or nitrate then dried and sometimes smoked (if desired for certain
flavor characteristics).

Shelf-stable for the purposes of shelf-stable meat and poultry products is defined as
the condition achieved when meat and poultry products can be stored under ambient
temperature and humidity conditions; if the package integrity is maintained during
storage, shipping, and display at retail and in the home; and the product will not spoil or
become unsafe throughout the manufacturer’s specified shelf-life.

Soudjouk: a highly spiced, Mediterranean-style, semi-dry sausage made by mixing

ground meat, spices, and curing salts, and then stuffing the resulting batter into a
natural or artificial casing that is flattened and subsequently fermented and dried at
ambient temperature over several days (Saricoban et al., 2006). Other spellings include
sujuk, sucuk, soujouk, and sudjuk.

Stabilization is the process of preventing or limiting the growth of spore-forming

bacteria capable of producing toxins either in the product before consumption or in the
human intestine after consumption. Establishments may use a variety different
103
stabilization processes, such as cooling, hot-holding, or meeting and maintaining certain
pH or water activity levels.

Summer sausage: refers to semi-dry sausages, especially Thuringer Cervelat. For

more information see the entry in Food Standards and Labeling Policy Book available
at: https://www.fsis.usda.gov/guidelines/2005-0003.

Targets: quantifiable pathogen reduction levels or growth limits set by the

establishment to produce safe products in the absence of regulatory performance
standards. As required by 9 CFR 417.2(c)(3), critical limits shall, at a minimum, be
designed to ensure that applicable targets or performance standards established by
FSIS, and any other requirement pertaining to the specific process or product be met.

Water activity: abbreviated as aw, water activity is a measure of the concentration of

moisture (i.e., water) and its availability in a food. The amount of water available in a
food depends on the total concentration of all dissolved substances in the product
because they bind water. Thus, if ingredients, such as salt or sugar, are added to food,
they compete with the bacteria for available water.

104
https://www.fsis.usda.gov/contact-us/askfsis

FSIS/USDA
www.fsis.usda.gov
2023

105