Sampling Requirements to Demonstrate Process Control in Slaughter Operations 

3/30/2020 

Sampling Requirements to Demonstrate Process

Control in Slaughter Operations 

Objectives 

After completion of this module, the participant will be able to: 

1. Explain why generic E. coli sampling and analysis is performed in livestock 

(other than swine) slaughter operations. 

2. Explain why microbiological sampling and analysis is performed in swine and 

poultry slaughter (other than ratite) operations. 

3. Identify who is responsible for selecting and analyzing livestock (other than 

swine) samples for generic E. coli. 

4. Identify who is responsible for selecting and analyzing swine and poultry 

samples for microbiological analysis. 

5. Explain the purpose of performance criteria and statistical process control. 

6. Describe how to verify the regulatory requirements for generic E. coli testing 

when conducting the Generic E. coli verification task. 

7. Describe how to verify the regulatory requirements for microbiological 

sampling and analysis of swine and poultry slaughter when conducting the 

appropriate PHIS inspection verification task. 

8. Explain the appropriate enforcement actions to take when noncompliance is 

found while performing the Generic E. coli verification task or the appropriate 

swine and poultry slaughter inspection verification task. 

References 

FSIS Directive 6410.4, Verifying Swine Slaughter Establishments Maintain 

Adequate Procedures for Preventing Contamination of Carcasses and Parts by 

Enteric Pathogens, 12/19/19 

FSIS Directive 6420.5, Verifying Poultry Slaughter Establishments Maintain 

Adequate Procedures for Preventing Contamination with Feces and Enteric 

Pathogens, 10/17/2016 

FSIS Directive 5000.1, Verifying an Establishment’s Food Safety System, Rev. 5, 

4/4/17 

Pathogen Reduction; Hazard Analysis and Critical Control Points (HACCP) 

System Final Rule (Federal Register Docket No. 93-016F; 1996) 

Modernization of Poultry Slaughter Inspection; Final Rule (Federal Register 

Docket No. FSIS-2011-0012, 2014) 

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Sampling Requirements to Demonstrate Process Control in Slaughter Operations 

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Modernization of Swine Slaughter Inspection; Final Rule (Federal Register 

Docket No. FSIS–2016–001, 2019) 

9 CFR 310.18(c)&(d), Contamination of carcasses, organs, or other parts. 

(Swine) 

9 CFR 310.25(a), Contamination with microorganisms; process control 

verification criteria and testing; pathogen reduction standards. (Livestock other 

than swine) 

9 CFR 381.65(g)&(h), Operations and procedures, generally. (Poultry) 

9 CFR 381.94, Contamination with microorganisms; process control verification 

criteria and testing; pathogen reduction standards for establishments that 

slaughter ratites. 

9 CFR Part 416 Sanitation SOP requirements 

9 CFR Part 417 HACCP requirements 

Guidelines for Escherichia coli Testing for Process Control Verification in Cattle 

and Swine Slaughter Establishments, FSIS Website: 

https://www.fsis.usda.gov/wps/wcm/connect/cebac8d0-954b-4cc2-abcf205ad4f4ddcd/Guideline_for_Ecoli_Testing_Cattle_Swine_Estab.pdf?MOD=AJP 

ERES 

FSIS Compliance Guidelines for the Control of Salmonella and Campylobacter in 

Raw Poultry, FSIS Website: 

https://www.fsis.usda.gov/wps/wcm/connect/6732c082-af40-415e-9b57-

90533ea4c252/Controlling-Salmonella-Campylobacter-Poultry2015.pdf?MOD=AJPERES 

Small and Very Small Plant Outreach 

https://www.fsis.usda.gov/wps/portal/fsis/topics/regulatorycompliance/haccp/small-and-very-small-plant-outreach/svsp-outreach 

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Introduction and History 

In 1996 the Pathogen Reduction; Hazard Analysis and Critical Control Points 

(HACCP) System Final Rule required all livestock and poultry slaughter 

establishments to test for generic E. coli to verify their process controls for 

microbiological contamination. Process control is defined as those activities that 

ensure a production process is stable and consistently operating at the desired 

level of performance, allowing for normal variation. This rule required 

establishments to develop, implement, and maintain written procedures to 

prevent microbiological contamination of carcasses. 

FSIS published the Modernization of Poultry Slaughter Inspection Final Rule in 

2014, and in 2019 the Modernization of Swine Slaughter Inspection Final Rule. 

These regulations expanded on the previous rule. Swine and poultry slaughter 

establishments must develop, implement, and maintain written procedures to 

prevent contamination of carcasses by enteric pathogens and fecal material 

throughout the entire slaughter and dressing operation. Enteric pathogens are 

bacteria associated with the intestine that can cause illness, for example, 

Salmonella. Each establishment must incorporate these written procedures, 

including their microbiological sampling plans, into their HACCP system, that is, 

the HACCP plan, or Sanitation SOP, or other prerequisite program. These 

procedures must include sampling and analysis for microorganisms, however, 

rather than requiring testing specifically for generic E. coli, swine and poultry 

establishments can choose any appropriate indicator organism. 

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Generic E. coli Testing for Livestock, (other than swine) 

Each official establishment that slaughters livestock, other than swine, or ratites 

is required to test for Escherichia coli Biotype I, also known as “generic E. coli.” 

An establishment employee selects the samples for generic E. coli testing. The 

purpose of generic E. coli testing is to verify the effectiveness of sanitation and 

process control in slaughter establishments. FSIS verifies that the establishment 

meets the regulatory requirements for generic E. coli testing. 

NOTE: Ratite is the group of birds that includes ostrich, rhea, and emu. These 

birds are amenable to inspection. Ratites are large, and when slaughtered, are 

treated like livestock. Although this section will focus on livestock, the regulations 

for ratites, 381.94(a), are included in the back of this module. 

NOTE: This section focuses on livestock, other than swine, because swine is 

covered in the next section. 

Fecal contamination is one of the principal sources of pathogenic organisms that 

contaminate livestock carcasses. Generic E. coli is an indicator of fecal 

contamination because it is common in the intestinal tract of food animals. The 

intestinal tract is also the primary pathway for contamination of carcasses with 

pathogens such as E. coli O157:H7, and Salmonella. Ongoing E. coli testing by 

livestock slaughter establishments helps them determine whether the slaughter 

process is under control or whether carcasses are being contaminated with 

feces. In other words, generic E. coli testing is a process control indicator for 

fecal contamination. Section 310.25(a) of the meat regulations addresses the 

regulatory requirements that establishments need to meet for generic E. coli 

testing. 

Slaughtered livestock that will not receive the FSIS mark of inspection (such as 

custom exempt livestock) are exempt from generic E. coli testing. 

Performance Criteria 

FSIS has developed performance criteria for beef using the excision sampling 

technique. Generic E. coli performance criteria are not enforceable regulatory 

standards. Performance criteria are numbers published in the regulations that 

represent the highest expected microbial loads on carcasses when the slaughter 

process is under control. They give livestock slaughter establishments guidance 

about the effectiveness of their slaughter process in preventing fecal 

contamination. Test results that meet the criteria in the regulations provide 

evidence that the establishment is maintaining adequate process control for fecal 

contamination and sanitary dressing. 

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Furthermore, the generic E. coli baseline results (statistical process control 

criteria) published in the Federal Register Notice on February 17, 2005 (Docket 

Number 02-046N), using the sponging sampling technique, can serve as a 

valuable support to establishments that slaughter cattle in assessing the 

effectiveness of their process, using their own test results. 

NOTE: Establishments must use statistical process control to evaluate their test 

results when they slaughter species or use sampling techniques for which the 

Agency has not developed performance criteria. 

Inspection Program Personnel (IPP) Responsibilities 

FSIS responsibilities are outlined in FSIS PHIS Directive 5000.1. IPP are 

responsible for understanding and properly performing the Generic E. coli 

verification task in the Public Health Information System (PHIS) as described in 

this directive. The Generic E. coli verification task addresses the regulatory 

requirements of 9 CFR 310.25(a) that the establishment must meet when 

developing the written generic E. coli testing procedure. 

IPP are to perform the Generic E. coli verification task on a routine basis (priority 

scale level 6) at the frequency specified in the establishment’s task list. IPP are 

also to initiate a directed Generic E. coli verification task if they observe 

noncompliance with the generic E. coli testing requirements while performing 

other tasks or when instructed to do so by supervision or other policy issuances. 

Generic E. Coli Testing Verification 

Establishments that slaughter livestock (other than swine) must develop a written 

sampling procedure that identifies the employees designated to collect samples, 

the locations of sampling, how randomness is achieved, and measures to ensure 

sample integrity as described in 9 CFR 310.25(a)(2)(i). 

IPP verify that the establishment meets the applicable regulatory requirements 

for generic E. coli testing by reviewing the establishment’s written sampling 

procedure, observing the designated establishment’s employee executing the 

written sample procedures and reviewing the establishment’s records. IPP are to 

document the results of their tasks in PHIS, including any noncompliance, 

according to the instructions described in FSIS Directive 5000.1. 

E. coli testing requirements are met if the establishment successfully executes 

the activities addressed in its written procedure, analyzes samples, and keeps 

records of test results. See the E. Coli Testing Summary Chart (Attachment 1) for 

an inspection aid to conducting the Generic E. coli verification task. 

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Sampling Requirements to Demonstrate Process Control in Slaughter Operations 

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IPP must understand what each section of the regulation means in order to 

conduct the Generic E. coli verification task. 

1. Sample collection – livestock 

E. coli testing must be done in establishments that slaughter any market class of 

cattle, sheep, goats, or equines. 

If a combination of types of livestock is slaughtered, the establishment samples 

only from the species it slaughters in the largest number. It is only necessary to 

sample one type of livestock to determine whether sanitary dressing controls are 

effective. E. coli tests measure the effectiveness of the process regardless of 

which species is slaughtered. This means, for example, if an establishment 

slaughters both cattle and sheep, but mostly cattle, they should be testing cattle 

for generic E. coli. 

NOTE: IPP are to judge which type of livestock is slaughtered in the greatest 

numbers based on historical slaughter numbers, for example, the previous year’s 

totals. 

Workshop 1: Generic E. coli Testing 

State the purpose of the generic E. coli testing regulations. 

What species of livestock are covered by the generic E. coli testing regulations? 

(§310.25) 

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2. Sampling requirements – location and technique 

The IPP should remember the following things when considering the sample 

location and technique. 

• The location refers to the place within the establishment where the sample 

is collected. 

• Livestock samples are collected after they have been in the cooler for a 

minimum of 12 hours. There is no maximum time limit. Carcasses can be 

selected while on the rail or after the final wash and set aside in a 

convenient spot in the cooler for testing after cooling. It is acceptable to 

select random samples before carcasses enter the cooler. 

• Hot-boning operation samples are taken after the final wash prior to 

boning. 

There are two sampling methods an establishment may use to collect generic E. 

coli samples. 

• Excision 

• Sponging 

Excision sampling is aseptically cutting a surface section from the livestock 

carcass and sending the tissue sample for laboratory analysis. Excising tissue 

from a carcass is, of course, a destructive method of sampling. 

Sponging is aseptically swabbing the surface of the livestock carcass with a 

sterile sponge and sending the sponge to the laboratory for analysis. Sponging is 

a nondestructive method of sampling. 

The chart below provides an easy reference for species and the sampling 

methods allowed. 

Excision Sponge 

Beef Beef 

Equine 

Goats 

Sheep 

Ratites 

Notice that beef carcasses may be sampled by the excision or sponging method. 

The sampling site refers to places on the carcass where samples are collected. 

Samples must be taken from specific sites on livestock carcasses. The three 

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sites from which excision samples on cattle or sponge samples on cattle, sheep, 

goat, and equine carcasses must be taken are the: 

• Flank 

• Brisket 

• Rump 

In the case of hide-on carcasses for the above species, the sponge samples 

must be taken from: 

• Inside the flank 

• Inside the brisket 

• Inside the rump 

NOTE: No excision samples are taken from hide-on carcasses. 

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Workshop 2: Carcass Sampling Sites 

1. In the left column of species, enter the matching letter for the regulatory 

sample sites listed in the right column. 

_____ Cattle A. Flank, brisket, rump 

_____ Goats B. Inside flank, brisket, rump 

_____ Hide on calves 

_____ Hide on sheep 

2. What are the two sampling methods? 

3. For which sampling method does FSIS regulations include performance 

criteria? 

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Sampling Requirements to Demonstrate Process Control in Slaughter Operations 

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3. Sample requirements – frequency 

For E. coli testing purposes, livestock slaughter establishments are divided into 

two categories: very low volume establishments (VLV) and greater than very low 

volume establishments (>VLV). The categories of establishments are based on 

the establishment’s annual slaughter volume. 

Very low volume establishments are described as follows (310.25(a)(2)(v)): 

• Cattle, goats, sheep, horses, or other equine: Annually slaughter fewer 

than 6,000 head 

• Livestock combination: Annually slaughter fewer than a combination of 

6,000 cattle, plus sheep, goats, horses, or equines that equal no more 

than 20,000 animals total 

Very low volume establishments begin sampling the first full week they operate 

after June 1st. They continue collecting at least one sample per week in each 

week they operate until 13 samples are completed. The series of 13 tests must 

show process control before the series can be ended. If the 13th test indicates 

that the sanitary dressing process is out of control, the establishment must 

continue to test until process control is regained. 

Sampling over a period of time provides a better indication of the process control 

of the establishment than taking all samples at once. This means the 

establishment should not try to collect all 13 samples in the same day, or even in 

one week. 

Seasonal VLV operations must complete all E. coli testing during whichever 

months it operates. For example, a seasonal goat slaughter establishment that 

operates from September through December must begin testing during its first 

full week of operations and complete 13 tests before operations end in 

December. 

When a VLV establishment that has completed 13 tests for the year makes a 

major change that affects how well the process control measures works, weekly 

testing must be resumed until another series of 13 tests can establish the 

effectiveness of the changed process. 

Greater than very low volume establishments are those slaughtering more than 

the numbers indicated above for VLV establishments (paragraph (a)(2)(iii)). 

Greater than very low volume establishments slaughtering cattle, sheep, goats, 

horses, or equines must test 1 per 300 carcasses frequencies or a minimum of at 

least once per week, whichever is greater. 

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Slaughter volume does not always match frequency rates in the regulations. 

Establishments should account for extra slaughter volume. This can be done by 

conducting additional tests. For example, a cattle slaughter establishment that 

slaughters 450 cattle per day should test at least once a day. However, the 

remaining 150 carcasses should also be accounted for to monitor process 

control. To account for the extra slaughter volume, the establishment could “carry 

over” the 150 extra carcasses to the next day’s volume and conduct two E. coli 

tests on the second day. 

Establishments may substitute an alternative testing frequency for the one in the 

regulations by including E. coli testing in their HACCP plan (paragraph (a)(2)(iv)). 

The alternative frequency must be part of the establishment’s verification 

procedures for its HACCP plan. It may not change the regulatory performance 

criteria, or the limits determined by statistical process control. 

4. Sample requirements – random selection of carcasses 

For generic E. coli testing the regulations require that livestock carcasses for 

sampling be selected at random (paragraph (a)(2)(i)). Random means that every 

carcass should have an equal chance of being selected. Different methods, like 

random number tables or computer-generated random numbers may be used. 

Whatever the establishment chooses to use must be written into the E. coli 

sampling procedure. 

The random method selected by the establishment and written into its plan must 

be followed. The designated establishment employee must be familiar with the 

written random sampling method. 

In cattle, each half-carcass represents one unit eligible for sampling. Both the 

“leading” and “trailing” sides of a carcass should have an equal chance of being 

selected within the designated time frame. In other livestock species, each whole 

carcass represents one unit eligible for sampling. 

If more than one shift is operating at the establishment, the sample can be taken 

from either shift, provided the sample selection time is based on the appropriate 

sampling frequency. 

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Sampling Requirements to Demonstrate Process Control in Slaughter Operations 

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Sample analysis 

Some establishments conduct their own analyses. FSIS assumes 

establishments following the "Guidelines for E. coli Testing for Process Control 

Verification in Cattle and Swine Slaughter Establishments" will conduct their 

sampling in a manner that does not jeopardize the integrity of the sample or the 

reliability of the test results. Because these guidelines are not regulatory 

requirements, the establishment may choose to use a comparable sampling 

technique. 

Establishment laboratory employees might have a copy of the Association of 

Official Analytical Chemists (AOAC) procedures or articles from peer-reviewed 

scientific journals that describe their procedure. 

IPP are to review the establishment’s written programs and records to verify that 

the laboratory analyzes the samples using an AOAC Official Method or another 

method that meets the criteria in paragraph (a)(3) of 9 CFR 310.25. IPP are to 

determine whether the establishment has documentation to demonstrate that the 

laboratory method meets these criteria. When in doubt about whether a testing 

procedure is acceptable, IPP should go through the supervisory channels to ask 

the District Office for assistance. 

5. Records of test results 

Establishments are required to keep a table or a chart of the results for at 

least the most recent 13 test results. IPP should consider the length of 

operations. In cases where the establishment has not been operating long 

enough to have 13 test results, there is not noncompliance for a lack of testing. 

Generic E. coli tests are reported as a quantity or bacterial concentration. 

Bacterial concentration can be reported using either the Colony Forming Unit 

(CFU) or the Most Probable Number (MPN) based laboratory methods of 

analysis to evaluate the generic E. coli testing. These methods provide an 

estimate of the number of unit viable cells per sample. 

An establishment using the regulatory “m” and “M” criteria must record each 

test result in terms of colony forming units per square centimeter (CFU/cm2) for 

excision. 

IPP should match the units of measure with the testing technique used to ensure 

that results are reported correctly. They are to verify that the establishment 

records the results on a process control chart or table that shows at least the 

most recent 13 test results. 

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Establishments must keep records of the tables and charts with generic E. coli 

test results for 12 months. Establishments are not required to maintain a file of 

laboratory reports received from either an in-house laboratory or an outside 

laboratory. 

6. Criteria for evaluation of test results 

If the Agency does not have performance criteria published for the species being 

sampled or for the sampling technique being used, the establishment must use 

statistical process control values to document generic E. coli test results 

(paragraph (a)(5)(ii)). The only species and sampling technique that FSIS 

currently has performance criteria for is cattle, using excision. This means that 

whenever the sponge method is used, the establishment must use statistical 

process control. Statistical process control is the use of statistical methods, 

such as control charts, to evaluate the variability of a process. The purpose of 

statistical process control is to maintain control over the variability of the process, 

to improve some aspect of the product being produced. In this case, it is used to 

measure and manage the microbiological contamination of carcasses. 

Using Statistical Process Control (SPC) to Evaluate Test Results 

Statistical Process Control (SPC) for generic E. coli is required with products that 

were not represented by the PR/HACCP Rule by a performance standard, 

because no relevant baseline studies were available at the time. The generic E. 

coli results published in the Federal Register Notice (2005) can complement SPC 

by providing establishments with an additional measure of process control. The 

results below are for cattle carcasses sampled using the sponge method of 

sample collection. 

Class of Method 80th 98th 

product percentile percentile 

Cattle 

carcasses 

sponge 0.0 CFU/cm2 3.1 CFU/cm2 

SPC provides a powerful mechanism for establishments to monitor and interpret 

the data collected for ongoing HACCP verification. SPC can provide 

establishments with an early warning that their process may not be functioning as 

designed. This warning can allow establishments to make process modifications 

to bring their process back into control without failing the desired performance. 

There are no regulatory requirements for how statistical process controls are 

determined. Companies may use a variety of valid methods to determine limits 

for statistical process control. For example, establishments may calculate their 

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own statistics, hire a consultant company, or use a software package to develop 

statistical process control values. Once the establishment determines the 

process control values and has set generic E. coli criteria to define process 

control, and as long as the data points on the company chart stay within the 

control limits set by the company, the process is considered in control. 

An example of a method a company may use to develop a SPC program is as 

follows. The establishment: 

• Conducts a series of preliminary generic E. coli tests during operations 

• Charts the results in cfu/cm2 

• Collects test results long enough to have a true picture of its performance 

(about 30 days usually). 

• Determines the typical range of generic E. coli counts found normally 

• Sets upper and lower control limits based on test results 

How to Verify Statistical Process Control 

IPP are to verify that the establishment is evaluating the test results using 

statistical process control techniques. In this context, IPP are to verify that an 

establishment that uses statistical process control has assessed the historical 

―normal performance of the slaughter process when it was in control, and 

developed criteria that will indicate when the process may not be in control. IPP 

are to verify that the establishment uses generic E. coli testing results to identify 

times when the slaughter process is trending toward a loss of control and takes 

necessary actions to reestablish control. IPP are not to focus on the particular 

method the establishment uses to set process control criteria. Instead, they are 

to review the generic E. coli testing results and verify that the establishment has 

set generic E. coli criteria to define process control and responds to results 

outside those criteria. 

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Example Statistical Process Control chart 

The following example of a SPC chart plots test results in terms of test number, 

along the horizontal X-axis, against cfu/cm2 on the Y-axis. This livestock 

slaughter establishment set a centerline value for its process control, which 

indicates the center point of the acceptable range of test results. The upper 

control limit (UCL) line marks the highest test result value considered acceptable 

by the company. The test result shown at test number 6 is above the upper 

control limit. The company recognized that this result was probably due to a 

variation in its process that needed to be identified, eliminated, and prevented 

from recurring. According to the chart, the establishment measures were 

effective because the following test result was back in the acceptable range. 

-20 

0 

20 

40 

60 

80 

100 

120 

140 

1 3 5 7 9 11 13 15 17 

E. coli cfu/cm2 

Test Number 

E. coli Control Chart 

UCL 

X 

X 

X 

X 

X 

X 

X 

X 

X 

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Using Regulatory Performance Criteria (m/M Values) to Evaluate Test 

Results 

Cattle establishments that choose excision of three sites must use the m/M 

performance criteria published in the regulations for evaluating test results when 

they are available. Regulatory m/M criteria apply only to cattle sampling when the 

excision sampling technique is used. When performance criteria are published in 

the regulations, the E. coli test results are compared to the regulatory criteria and 

may fall into one of three categories: acceptable, marginal (represented by “m”), 

and unacceptable (represented by “M”). 

• Marginal results (“m”) are those that fall within the worst 20% of overall 

industry performance in terms of E. coli counts (results taken from 

baseline study). More than three marginal results in the last 13 tests are 

unacceptable. 

• Results in the worst 2% of overall industry performance (results taken 

from the baseline study) are called the maximum or “M” value. Any single 

test result exceeding “M” is unacceptable. 

The m/M values taken from the regulations are applied to a moving window of 

the last 13-documented test results. That means that the establishment 

considers all of the last 13 test results when determining if the process is in 

control. Every time a new test result is added to their records, the oldest test is 

dropped, and the new test becomes one of the most recent 13 results. 

For the slaughter process to be judged in control no more than three sample 

results can be above the “m” marginal line. If four sample results are above “m”, 

the process is out of control. 

If the test result of the most recent sample is above “M”, the process is 

automatically out of control, regardless of the previous test results. Once another 

test result is entered in the chart or table, the “M” test simply becomes another 

result considered to be above the “m” line. It no longer carries the consequence 

of causing “automatic” process control failure. 

After the slaughter process is judged to be out of control, a subsequent test result 

below the “m” line indicates that the establishment did something to correct a 

problem and bring the process back into control (this correction does not have to 

be documented anywhere). However, the process is not judged totally in control 

until the window of 13 tests also shows process control. 

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The following table from the regulations shows the m/M values for E. coli 

performance criteria set by the Agency for excision testing of cattle. 

Species Lower limit of 

marginal range 

(m) 

Upper limit of 

marginal range 

(M) 

Number of 

sample 

tested (n) 

Maximum # 

permitted in 

marginal 

range (c) 

Cattle Negative 100 CFU/cm2 13 3 

An example of how to use the table is to consider a cattle slaughter 

establishment that uses the excision sampling method. An E. coli test result is: 

• Acceptable if it comes back negative 

• Marginal if the test result is positive but not above 100 cfu/cm2 

• Unacceptable if it is above 100 cfu/cm2 

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The following table is an example of one method that may be used by 

establishments for record keeping. 

Cattle Excision Test Results 

Test 

Num. Date 

Test 

Result 

(cfu/cm2) 

Result 

unacceptable? 

Result 

marginal? 

Number 

marginal or 

unacceptable 

in last 13 

Pass/Fail? 

1 10-07 10 No Yes 1 Pass 

2 10-07 Negative No No 1 Pass 

3 10-08 50 No Yes 2 Pass 

4 10-08 Negative No No 2 Pass 

5 10-09 Negative No No 2 Pass 

6 10-09 Negative No No 2 Pass 

7 10-10 80 No Yes 3 Pass 

8 10-10 Negative No No 3 Pass 

9 10-11 Negative No No 3 Pass 

10 10-11 Negative No No 3 Pass 

11 10-14 50 No Yes 4 Fail 

12 10-14 Negative No No 4 Fail 

13 10-15 Negative No No 4 Fail 

14 10-15 Negative No No 3 Pass 

15 10-16 Negative No No 3 Pass 

16 10-16 Negative No No 2 Pass 

17 10-17 120 Yes No 3 Fail 

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Looking at this establishment record the following determinations can be made. 

1. Test number eleven, conducted on October 14th, documents the fourth test 

result in the marginal (“m”) range. Therefore, the establishment was in an 

unacceptable process control status because the fourth marginal result exceeds 

the limit of no more than three marginal results in the past 13 consecutive tests. 

IPP should focus on dressing procedures and sanitation performance standard 

requirements when failing test results indicate the lack of slaughter process 

control. 

2. Tests number twelve and thirteen are negative, and therefore in the acceptable 

range. However, considering the last 13 test results in the 13-test moving 

window, there are still more than three results in the marginal range. The 

company marked its record to show that it is still failing because there are four 

marginal test results. In reality this is not an unacceptable result because tests 

twelve and thirteen are negative, indicating the process is back in control, but 

there is evidence of problems in the recent past. 

3. For test number fourteen the number of marginal results in the last thirteen tests 

window is reduced to three. The marginal result for test number one is dropped 

and replaced by an acceptable result as the 13-test window moves ahead one 

test. 

4. The test result for test number seventeen exceeds 100 cfu/cm2, the “M” value for 

cattle. Any result over 100 cfu/cm2 is automatically unacceptable. It takes only 

one test in the “M” range to indicate the establishment may not have adequate 

process control. 

Inspection personnel reviewing this record should focus on sanitation 

performance standard requirements. 

Another method the company may use to document its E. coli test results is a control 

chart. The seventeen test results written in the previous table are plotted on the 

following control chart. 

The vertical Y-axis shows how many colony-forming units (cfu) of E. coli were found in a 

square centimeter (cm2) of the test sample analyzed at the laboratory. The horizontal 

X-axis indicates the test number. Marking an “X” at the point where the X and Y-axes 

converge (meets) reflects the test value or result for the particular test number. For 

ease of reading, the chart has a line to indicate the bottom limit of “m”, and a thicker line 

to indicate the upper limit of “M.” Any “X” plotted between the thin line and the thick line 

falls in the marginal range, we call “m.” Any “X” plotted above the thicker line is in the 

unacceptable range, or “M.” 

Inspection Methods 29-19 
E. Coli Control Chart 

Figure 4 

140 

120 . 

. 

§100 

C" 

en 80 .. 

~ 

-::::J 

-(.) 60 

)( )( 

0 40 

(.) 

w 20 

)( 

0 .. .. ... .. .. .. .. .. -

-20 

1 2 3 4 5 6 7 8 9 10 111213 14 15 16 17 

Test Number 

___ m 

M >< Test Resu It 

 Sampling Requirements to Demonstrate Process Control in Slaughter Operations

3/30/2020

Action to Regain Process Control

Whenever a prudent livestock slaughter establishment determines that its E. coli test 

results do not meet m/M performance criteria or statistical process control values, it 

should take necessary actions to bring the slaughter process back into control. 

Under the regulations, establishments are not required to take corrective actions or to 

document the necessary actions for E. coli test failures. However, when livestock 

slaughter establishments do not evaluate their test results (§310.25(a)(5)), they might 

not be maintaining slaughter process controls sufficient to prevent fecal contamination. 

Inspection Methods 29-20
 

 

Sampling Requirements to Demonstrate Process Control in Slaughter Operations 

3/30/2020 

7. Sample Integrity 

According to this section of the regulations, sample integrity must be addressed in the 

establishment’s written sample collection procedure and should be followed; but if it is 

not followed, it is not an enforceable issue. If IPP observe circumstances that seem to 

jeopardize sample integrity (e.g., freezing the sample, not shipping the sample on the 

same day it is collected), the District Office should be notified through supervisory 

channels. Further investigation of the situation and any enforcement actions will be 

directed from the District Office. 

Workshop 3: True or False 

____ 1. As per 9CFR 310.25(a)(2), establishments that slaughter livestock are only 

categorized as very low volume. 

____ 2. For generic E. coli testing, the regulation requires that livestock carcasses be 

selected at random for sampling. 

____ 3. It is not necessary for the establishment to keep a table or chart showing the 

most recent 13 test results. 

____ 4. If a cattle slaughter establishment uses the sponge sampling method, they must 

use the performance criteria published in 9 CFR 310.25 to evaluate generic E. coli test 

results. 

____ 5. Establishments that slaughter livestock must keep records of the tables or 

charts with generic E. coli test results for 12 months. 

Inspection Methods 29-21 
 

 

 

 

 

 

 

Sampling Requirements to Demonstrate Process Control in Slaughter Operations 

3/30/2020 

Microbiological Sampling for Poultry (other than Ratite) and Swine 

Slaughter Operations 

The purpose of the modernized sampling requirements is to ensure that establishments 

monitor and evaluate the effectiveness of their procedures to prevent contamination of 

carcasses by enteric pathogens and visible fecal material on an ongoing basis. Fecal 

contamination is a principal source of pathogenic organisms that contaminate poultry 

and swine carcasses. Under the Modernization of Poultry Slaughter Inspection and 

the Modernization of Swine Slaughter Inspection final rules establishments that 

slaughter poultry, other than ratites, and swine are required to perform microbiological 

sampling and analysis, for example, testing for Salmonella, Campylobacter, or indicator 

organisms such as aerobic plate count (APC), total coliform, Enterobacteriaceae, and 

Escherichia coli, Biotype I, also known as generic E. coli. 

Because establishments have differences in their operations, each establishment has 

the flexibility to develop a sampling plan and determine the microbial organism that will 

accurately monitor the effectiveness of its process control procedures. Establishments 

MUST incorporate their written process control procedures into their HACCP system, 

either in the HACCP plan itself, as sanitation SOPs, or as a prerequisite program. 

Microbiological test results that represent the level of microbiological contamination at 

key steps in the slaughter process are necessary for the establishment to provide 

comprehensive objective evidence to demonstrate process control. Process control 

consists of the programs and procedures that an establishment implements to ensure 

its process prevents contamination of poultry and swine carcasses, including 

contamination with pathogens and fecal material. Process control also ensures that the 

resulting product meets applicable standards or definitions. 

Inspection Program Personnel (IPP) Responsibilities 

Species Directive Regulation 

Poultry FSIS Directive 6420.5, Verifying Poultry 

Slaughter Establishments Maintain Adequate 

Procedures for Preventing Contamination 

with Feces and Enteric Pathogens, 

10/17/2016 

9 CFR Part 381.65(g)&(h) 

Swine FSIS Directive 6410.4, Verifying Swine 

Slaughter Establishments Maintain Adequate 

Procedures for Preventing Contamination of 

Carcasses and Parts by Enteric Pathogens, 

12/19/19 

9 CFR Part 310.18(c)&(d) 

Inspection Methods 29-22 
 

 

 

 

 

 

 

 

 

 

Sampling Requirements to Demonstrate Process Control in Slaughter Operations 

3/30/2020 

The PHIS verification task that IPP perform depends on how the establishment has 

incorporated its written procedures for preventing contamination of carcasses by enteric 

pathogens and fecal contamination throughout the entire slaughter and dressing 

operation in its HACCP system. For instance: 

• If the establishment’s written procedures are part of its HACCP plan, IPP are to 

verify HACCP regulatory requirements by performing the Slaughter HACCP 

verification task when it has been scheduled in PHIS. 

• If the establishment’s written procedures are part of its Sanitation SOPs, IPP are 

to verify that the establishment meets all Sanitation SOP regulatory requirements 

by performing the Operational SSOP Review and Observation task when it 

has been scheduled in PHIS. 

• If the establishment’s written procedures are part of another prerequisite program 

or other control measures, IPP are to verify the implementation of such program 

by performing the Slaughter HACCP verification task when it has been 

scheduled in PHIS. 

IPP are to perform the appropriate PHIS verification task on a routine basis at the 

frequency specified in the establishment’s task list. IPP are also to initiate a directed 

verification task if they observe noncompliance while performing other tasks or when 

instructed to do so by supervision or other policy issuances. Conduct HACCP and 

SSOP verification tasks as outlined in Directive 5000.1 and as covered in previous 

training modules. 

IPP are to verify that the poultry or swine slaughter establishment: 

• Developed a written sampling program that identifies the specific microorganisms 

being tested and location/frequency where samples are collected, 

• Incorporated its written sampling program for preventing contamination by enteric 

pathogens into its HACCP system, 

• Implements and maintains its written sampling program, 

• Maintains scientific and technical documentation to support the decisions that the 

establishment made in designing the sampling program, 

• Maintains daily records documenting the implementation and monitoring of its 

procedures including sample results 

• Take actions to restore or improve process control when sample results indicate 

problems with establishment slaughter HACCP system. 

Inspection Methods 29-23 
 

 

 

 

 

 

 

 

 

Sampling Requirements to Demonstrate Process Control in Slaughter Operations 

3/30/2020 

Microbiological Sampling and Analysis Verification 

Each poultry or swine slaughter establishment’s written procedures for preventing 

contamination of carcasses and parts with enteric pathogens and fecal material must 

include sampling and analysis for microbial organisms. 

The regulations require each establishment to maintain scientific and technical 

documentation to support the judgments that the establishment made in designing the 

sampling program. The regulations prescribe the minimum requirements for the location 

and frequency of sampling, based on the establishment size and production volume. 

Each establishment must maintain daily records to document the implementation and 

monitoring of their procedures including records documenting the test results of its 

sampling plan. 

Note: Poultry establishments may use Salmonella Initiative Program (SIP) microbial 

data as part of their sampling plan to monitor their process control, provided they meet 

minimum frequencies and location requirements. 

Summary charts (Poultry Attachment 2, and Swine Attachment 3 of this handout) are 

provided as a reference for the establishment size, sampling frequencies, and sampling 

locations requirements. They are a quick and easy inspection aid when conducting the 

PHIS verification tasks. 

IPP must understand what the regulation means in order to conduct the appropriate 

PHIS verification task. 

1. Sampling requirements – Microbial Indicator Organism 

Each establishment must develop its own sampling program/procedure that identifies 

the specific microbiological organisms (i.e., Salmonella, Campylobacter, or other enteric 

organisms) for which the establishment will test to monitor the effectiveness of its 

process control procedures that prevent contamination of carcasses and parts with 

enteric pathogens and fecal material. 

Note: An establishment may continue to test for generic E. coli, as was previously 

required, if the establishment determines such testing is effective for monitoring process 

control. 

2. Sampling requirements – Location and Frequency 

The location refers to the place within the establishment where the sample is collected. 

Frequency refers to how often samples must be taken. The required location and 

frequency depend on species and size of establishment. 

Inspection Methods 29-24 
 

 

 

 

Sampling Requirements to Demonstrate Process Control in Slaughter Operations 

3/30/2020 

Poultry slaughter establishments are codified by size and annual slaughter volume, 

according to regulation 381.65(g)(1)(i) and (ii). 

• Very small establishments are establishments with fewer than 10 employees or 

annual sales of less than $2.5 million. 

• Very low volume (VLV) establishments annually slaughter no more than 440,000 

chickens, 60,000 turkeys, 60,000 ducks, 60,000 geese, 60,000 guineas, 60,000 

squabs or a combination of all types of poultry not exceeding 60,000 turkeys and 

440,000 birds total. 

Very small poultry establishments and VLV establishments operating under Traditional 

inspection are required to collect samples for microbial organisms at the post-chill 

point in the process. All other establishments must collect samples at both the pre-chill 

and post-chill locations. 

Traditional Poultry - VS 

& VLV 

All Other Poultry 

Establishments 

Post-chill only Pre-chill 

and 

Post-chill 

The pre-chill location for sampling is any point in the poultry slaughter process from rehang to just prior to the chiller. The post-chill location for sampling is a point in the 

slaughter process after the carcass exits the chiller and after all slaughter interventions 

are completed. 

Poultry VLV establishments must collect and analyze samples at least once during each 

week of operation starting June 1 of every year, per 9 CFR 381.65(g)(2)(ii). If, after 

consecutively collecting 13 weekly samples, a VLV establishment can demonstrate that 

it is effectively maintaining process control, it may modify its sampling plan. 

All other poultry establishments (including very small establishments) must collect and 

analyze a pair of samples per 9 CFR 381.65(g)(2(i), one at pre-chill and one at postchill, at the following frequencies: 

• Chickens: once per 22,000 carcasses but at a minimum of once during each 

week of operation; 

• Turkeys, ducks, geese, guineas, and squabs: once per 3,000 carcasses but at a 

minimum once each week of operation. 

Poultry carcasses must be selected at the required points in the process (pre and post 

chill). At the post-chill site, samples should be collected after the final wash and the 

application of any final antimicrobial interventions. A drip time of at least 60 seconds 

Inspection Methods 29-25 
 

 

 

 

Sampling Requirements to Demonstrate Process Control in Slaughter Operations 

3/30/2020 

should be observed before sample collection to prevent excessive antimicrobial 

carryover in the collected sample. 

Note: Antimicrobials used during processing steps may make it harder to detect live 

bacteria in the collected sample if the carcass is not allowed adequate drip time before 

collecting the sample. Consequently, antimicrobial carryover (residual) can result in 

altered test results (lower bacterial counts), may invalidate the test results, and may not 

provide a true representation of the establishment’s process control. 

The sampling methods for collecting poultry carcass samples may include the 

nondestructive sponge technique for sample collection from turkeys and geese (back 

and thigh) and a whole bird rinse technique for sample collection from chickens, 

guineas, ducks, geese, and squabs. All carcass samples should be taken using aseptic 

techniques. 

Swine slaughter establishments are codified by annual slaughter volume, according to 

regulation 9 CFR 310.18(c)(1)(i). A very low volume (VLV) establishment annually 

slaughters no more than 20,000 swine, or a combination of swine and other livestock 

not exceeding 6,000 cattle and 20,000 total of all livestock. 

Very low-volume swine slaughter establishments must collect and analyze at least one 

post-chill sample per week of operation starting June 1 of each year. If after testing 13 

consecutive weekly samples, a very low-volume swine establishment can demonstrate 

that it is effectively maintaining process control, the establishment can discontinue 

sampling for the year. 

All other swine establishments must collect a set of two samples per every 1,000 

carcasses, one at pre-evisceration and one at post-chill. Swine establishments that 

“hot-bone” should collect the two samples at pre-evisceration and after the final wash 

location. The two samples do not need to be taken from the same carcass. 

Very Low-Volume 

Swine 

All Other Swine 

Establishments 

Post-chill only Pre-evisceration 

and 

Post-chill 

All swine establishments must sponge or excise tissue from the ham, belly, and jowl 

areas, per 9 CFR 310.18(c)(1). 

All establishments must provide scientific or technical support for their sampling 

technique and sample site on the carcass. If IPP have concerns with the 

establishment’s support, they should contact the District Office through supervisory 

channels. 

Inspection Methods 29-26 
 

 

 

 

 

 

 

 

 

 

Sampling Requirements to Demonstrate Process Control in Slaughter Operations 

3/30/2020 

Random selection of carcasses 

Samples should be collected randomly at the frequency determined by the 

establishment as part of its sampling plan. If more than one shift is operating at the 

establishment, the sample can be taken on any shift. Different methods of selecting the 

specific carcass for sampling could be used, but the method used should include the 

use of random numbers to ensure that testing data is not biased. Examples of methods 

include random number tables, calculator or computer-generated random numbers, or 

drawing cards. 

The carcass that is sampled should be selected at random from all eligible carcasses. If 

there are multiple lines or chillers, randomly select the line or chiller for sample 

collection. 

Sample analysis and testing method 

To obtain the most accurate results, samples should be analyzed as soon after 

collection as possible. If samples must be transported to an off-site laboratory, they 

should be refrigerated and then shipped refrigerated, on the same day they were 

collected, via an express delivery service to the laboratory. A sample should arrive at 

the laboratory and be analyzed no later than the day after it is collected. 

Each establishment needs to determine whether sample analysis will be performed by 

an outside or on-site laboratory. FSIS has available the compliance guideline 

“Establishment Guidance for the Selection of a Commercial or Private Microbiological 

Testing Laboratory” if the establishment decides to use an outside laboratory to analyze 

microbiological samples. This guidance document should be particularly useful to very 

small establishments when they are selecting a commercial or private laboratory to 

analyze establishment microbiological samples. 

FSIS has also made available a list of Foodborne Pathogen Test Kits Validated by 

Independent Organizations for the detection of relevant foodborne pathogens. This list 

is guidance and is not an endorsement of any particular testing method. 

Poultry and swine slaughter establishments must include the analysis of microbial 

organisms in their sampling procedures as part of their HACCP system. Therefore, 

scientific and technical documentation must be provided to support the design of the 

sampling program. 

IPP are to review the establishment’s written programs, scientific and technical support, 

and records to verify that the laboratory analyzes the samples using an AOAC Official 

Method or one validated by another recognized independent testing body. When in 

doubt about whether the laboratory testing procedure is acceptable, IPP should go 

through the supervisory chain-of-command to the District Office for assistance. 

Inspection Methods 29-27 
 

 

 

 

 

 

T.albl,e 1 - rndi,cator Orga,111 ·sm Med·arn Values ifoli Chi1ckens 

Medlia1n {CF Ul'm Li 

Gernemic E. APC Enteroba,c·t,eriaoeae Total 

coli Col iform 

Ca1rcass - Rehan.a 540 28,000 1,600 '940 

Car cass. - Post 20 260 20 20 

Chnl 

Table 2 - llndlicat1or Organism Median Value.s foir Tuirk.eys 

Median 1€CFUJmU 

Gen,eri,c E. coli APC Enteroibact,eriaoeae Total Coliform 

Car cass - Rehan,g 22 1,800 50 40 

Carcass - P:ost' Chm <1 .2 18 <11.2 < 'L2 

Sampling Requirements to Demonstrate Process Control in Slaughter Operations 

3/30/2020 

Records of test results 

Poultry and swine slaughter establishments must maintain daily records documenting 

the implementation and monitoring of its procedures to prevent contamination of 

carcasses by enteric pathogens, including accurate records of all test results from its 

sampling plan, for at least one year. These records must be available for review by IPP 

upon request. 

IPP are to verify that the establishment maintains daily records documenting the 

implementation and monitoring of its procedures, makes these records available for 

review and retains these records for one year. 

Criteria for evaluation of test results 

Poultry and swine slaughter establishments should use statistically valid approach or 

statistical process control (SPC) to interpret their microbiological test results as 

previously discussed in this handout. 

In cases where an establishment does not have the resources or capacity to develop 

and implement their own statistical control limits or procedures, establishments can 

utilize the results from FSIS nationwide livestock or poultry surveys. 

The tables below demonstrate the indicator organism median values for chickens and 

turkeys. 

An establishment sample value that is higher than the corresponding one listed in the 

table indicates the establishment may not be maintaining process control and may be 

less likely to meet applicable performance standards. Sample values lower than the one 

listed in the table indicate the establishment may be maintaining process control. 

Inspection Methods 29-28 
 

 

 

 

 

 

0. 

0.6. 

0. I 

0.2 

0.0 

i(!I 

Ch am l - System under oontiroll 

• 

• • 

• 

• • 

5 10 l5 

Sampling Requirements to Demonstrate Process Control in Slaughter Operations 

3/30/2020 

SPC usually includes the use of a control chart, which plots data over time but also 

displays an upper control limit for specific measurements and a centerline (the 

average), above and below which there is an equal number of sample results. A sample 

result above the upper control limit would indicate the likely presence of a special cause 

of variation that should be addressed. Results within control limits indicate that the 

process is in control. 

The example below shows a SPC chart for a slaughter operation which plots test results 

for an indicator organism in terms of sample number, along the horizontal X-axis, 

against Log cfu/ml on the Y-axis. This chart illustrates a pattern of an indicator organism 

test results that would be seen in a well-controlled system. In a well-controlled system, 

the majority of the test results will be clustered around a central value (the average). It is 

important to note that even a well-controlled system there is some frequency of isolated 

results above the acceptable level. Log (cfu/ml) 

As part of its process control procedures, an establishment should define the actions it 

will take if the microbiological test results obtained through its sampling are above the 

limits it has set. The establishment should delineate what its actions will be, who will 

take each action, how the outcome of these actions will be documented, and how it will 

be verified. 

FSIS has made available the FSIS Compliance Guidelines for the Control of Salmonella 

and Campylobacter in Raw Poultry. The guidelines summarize known control points for 

Salmonella and Campylobacter in the pre- and post-harvest production process. 

Establishments should use this compliance guide to improve management practices, to 

ensure effective dressing operations and to assist in investigating when there is a loss 

of control of the slaughter process. 

Inspection Methods 29-29 
 

 

 

Sampling Requirements to Demonstrate Process Control in Slaughter Operations 

3/30/2020 

When IPP review the establishment’s records that document its microbiological test 

results, they should look for trends in the test results that indicate a loss of process 

control. For example, IPP are to look for: 

• A significant number of test results that exceeded the establishment’s upper 

control criteria, if the establishment has such criteria, 

• Instances where the test results exceed the establishment’s criteria by a large 

amount over a relatively short period of time (e.g., days or weeks); or 

• Test results that show a trend of worsening performance over a relatively long 

period of time (e.g., days, months, seasonal). 

• Other sampling plans begin to show significantly worse results. This could 

include FSIS positive pathogen sampling results. This may indicate increased 

contamination is occurring during slaughter. 

Sample Integrity 

A prudent establishment should include a description of how samples are handled to 

ensure sample integrity. The regulations require poultry and swine slaughter 

establishment to incorporate their written procedures in its HACCP system, either in its 

HACCP plan, its Sanitation SOPs, or another prerequisite program. Implementation of 

the program must then comply with the 9 CFR 416 or 417, depending on where the 

establishment chooses to locate it. 

Inspection Methods 29-30 
 

 

 

 

 

 

 

 

 

Sampling Requirements to Demonstrate Process Control in Slaughter Operations 

3/30/2020 

Documenting Inspection Results in PHIS 

IPP are to follow instructions for documenting their inspection results in PHIS as 

described in FSIS Directive 5,000.1. When the establishment is in compliance with the 

regulations, IPP select the mandatory regulations, any other regulation they verified on 

the “Regulations” tab and mark the task as ‘Inspection Completed’ at the bottom of the 

Inspection Results page. If IPP find noncompliance, they are to notify the establishment 

and document the noncompliance on an NR citing the appropriate regulation. 

Noncompliance in Livestock (other than swine) Slaughter Establishments 

The livestock (other than swine) slaughter establishment’s generic E. coli testing results 

cannot, by themselves, support a finding of noncompliance with 9 CFR 310.25(a) 

However, if the establishment’s testing results indicate a failure of process control, IPP 

are to verify the establishment’s sanitary dressing procedures. IPP should use the 

findings from verifying the establishment’s sanitary dressing procedures in conjunction 

with other information, like zero tolerance failures and positive E. coli O157:H7 results (if 

applicable) in beef slaughter processing combination establishments and any other 

HACCP performance, in making determinations regarding the effectiveness of the food 

safety system. 

Noncompliance occurs when the establishment is not meeting the prescribed regulatory 

requirements in 9 CFR 310.25(a) (refer to Directive 5,000.1, Chapter IV – Pathogen 

Reduction Activities). 

The following findings are evidence that the establishment does not comply with 9 CFR 

310.25(a) 

1. The establishment is not conducting sampling at the required location. 

2. The establishment is not using the required sampling technique or sampling at 

the required site on the carcass. 

3. The establishment is not sampling the required frequency according to the 

establishment’s production volume. 

4. The laboratory is not using a quantitative method for analysis of generic E. coli 

that is approved as an Official Method of the AOAC International or approved 

and published by a scientific body. 

5. Records are not available for FSIS access or not retained for 12 months 

Inspection Methods 29-31 
 

 

 

 

 

 

 

 

 

Sampling Requirements to Demonstrate Process Control in Slaughter Operations 

3/30/2020 

Noncompliance Poultry and Swine Slaughter Establishments 

The establishments test results by themselves do not necessarily indicate 

noncompliance. IPP are to consider all available information, including results from 

FSIS testing, to determine whether the establishment’s microbiological sampling 

program enables the establishment to monitor its ability to maintain process control, 

including how the establishment is implementing its sampling program, any trends that 

are occurring in the test results, and how the establishment is reacting to its test results. 

Noncompliance occurs when the establishment is not meeting the prescribed regulatory 

requirements; is not following its written sampling and testing procedures; does not 

demonstrate that it is maintaining process control; or its corrective actions are not 

effective. 

Poultry (Directive 6420.5) 

The following findings are evidence that the establishment does not comply with 9 CFR 

381.65(g) and 9 CFR 381.65(h) (refer to Directive 6,420.5, Section VII). 

1. The establishment has not addressed minimizing contamination by enteric 

pathogens and fecal contamination at steps along the line in the slaughter 

process. 

2. If the establishment is not implementing their written procedures. 

Note: Additional regulatory noncompliance may need to be cited on the NR 

depending on where the establishment has incorporated its written procedures in 

the HACCP system. For example, if the written procedures for preventing 

contamination by enteric pathogens and fecal material throughout slaughter and 

dressing operations are in the SSOP, IPP should also cite 9 CFR 416.13(b) and 

(c) when the establishment is not implementing those procedures. Likewise, if the 

written procedures for preventing contamination by enteric pathogens and fecal 

material throughout slaughter and dressing operations are in a prerequisite 

program, IPP should also cite 417.5(a)(1) when the establishment is not 

implementing those procedures. 

3. The procedures are not effective in preventing contamination, e.g., the system 

results in little or no reduction in visible fecal contamination, or the 

establishment’s microbiological test results indicate that the establishment’s 

procedures are not effective in maintaining process control. 

4. The establishment is not conducting microbiological sampling at the required 

location or at the required frequency according to the establishment’s size and 

production volume. 

Inspection Methods 29-32 
 

 

 

 

 

 

Sampling Requirements to Demonstrate Process Control in Slaughter Operations 

3/30/2020 

5. Records are not available for FSIS access or not retained for one year. 

6. Sample integrity, e.g. randomness and handling of samples, is not maintained. 

Note: If the establishment does not have written procedures to prevent enteric 

pathogen or visible fecal contamination throughout the slaughter process, or has not 

incorporated the procedures into its HACCP system, IPP are to perform the PHIS 

Slaughter HACCP verification task and issue a noncompliance record (NR) citing 9 CFR 

381.65(g), 381.65(h), 417.2(a), and 417.5. 

Swine (Directive 6410.4) 

1. The establishment does not have necessary support for its sampling plan to 

show that its testing is effective in determining whether the system is preventing 

pathogens (417.5(a)(1)). 

2. The establishment does not include support for testing for indicator organisms 

(417.5(a)(1)). 

3. The establishment is not, at a minimum, conducting microbiological sampling at 

the required location(s) or frequency according to the establishment’s size and 

production volume (9 CFR 310.18(c)). 

4. The establishment does not maintain sample integrity, (e.g., randomness and 

handling of samples) (9 CFR 310.18(c)). 

5. The establishment is not maintaining daily records to document the 

implementation and monitoring of its written procedures (9 CFR 310.18(d)). 

6. The establishment does not make records available for FSIS review or does not 

retain records for one year (9 CFR 310.18(d)). 

7. The establishment does not conduct corrective actions to address findings of 

visible fecal, ingesta, or milk contamination as required by HACCP (9 CFR 

417.3), Sanitation SOPs (9 CFR 416.15), or other prerequisite programs, 

described in FSIS Directive 5000.1 (9 CFR 417.5). 

Inspection Methods 29-33 
 

 

 

 

 

 

 

 

Sampling Requirements to Demonstrate Process Control in Slaughter Operations 

3/30/2020 

Enforcement 

Livestock (other than swine) Slaughter Establishments (Directive 5,000.1) 

FSIS generic E. coli criteria are guidelines, not regulatory standards. FSIS does not use 

company Generic E. coli test results to take regulatory action. Test results that show 

lack of process control should be considered in conjunction with other information, like 

sanitary dressing, SSOP and HACCP performance. 

Further enforcement action might be necessary if the establishment repeatedly fails to 

implement appropriate immediate action or further planned action in response to NRs 

documenting noncompliance. In these cases, the inspector in charge (IIC) should notify 

the District Office through supervisory channels. The District Office will give instructions 

for additional enforcement action when necessary. 

Poultry and Swine Slaughter Establishments (Directives 5,000.1, 6,420.5, and 

6410.4) 

IPP are to consider the following: 

• Their findings in combination with poultry or swine zero tolerance findings, 

together with other findings to evaluate whether the establishment is effectively 

implementing the components of its HACCP system to ensure that poultry are 

slaughtered under sanitary conditions. 

• Whether the overall pattern of inspection findings suggests that the 

establishment is not maintaining sanitary conditions throughout the slaughter 

HACCP system. For example: 

o an establishment has repetitive associated HACCP or SSOP 

noncompliance for multiple aspects of the slaughter system 

o the establishment’s corrective actions in response to findings of visible 

fecal contamination are consistently ineffective – may indicate systemic 

problems with its slaughter HACCP system. 

If the establishment has repetitive NRs, or the establishment’s corrective actions are 

ineffective, IPP are to discuss with their immediate supervisor the need to take an 

enforcement action outlined in FSIS Directive 5000.1., Chapter V. 

Inspection Methods 29-34 
 

 

Sampling Requirements to Demonstrate Process Control in Slaughter Operations 

3/30/2020 

Attachment 1 

GENERIC E. COLI (LIVESTOCK OTHER THAN SWINE AND RATITES) TESTING

SUMMARY CHART 

TEST TEST SAMPLE SAMPLING 

SPECIES FREQUENCY LOCATION SITES METHOD 

Cattle 1/300 

carcasses or 

1/wk. – 

whichever is 

greater 

Carcass cooler 

>12 hrs. 

Hot boned: after 

final wash 

Flank, brisket, 

rump 

Excision* 

Sponging 

Hide-on 

carcasses 

(calves, 

sheep, goats) 

1/300 

carcasses or 

1/wk. – 

whichever is 

greater 

Chilled 

carcasses 

Inside flank, 

inside brisket, 

inside rump 

Sponging only 

Horses, 

Mules, Other 

Equines 

1/300 

carcasses or 

1/wk. – 

whichever is 

greater 

Chilled 

carcasses 

Hot boned: after 

final wash 

Flank, brisket, 

rump 

Sponging 

Sheep and 

Goats 

1/300 

carcasses or 

1/wk. – 

whichever is 

greater 

Chilled 

carcasses 

Hot boned: after 

final wash 

Flank, brisket, 

rump Sponging 

Ratites 1/3000 

carcasses or 

1/wk. – 

whichever is 

greater 

Chilled 

carcasses 

Hot boned: after 

final wash 

Sponge back and 

thigh 

Sponging 

* Have applicable m/M values 

Inspection Methods 29-35 
 

 

 

 

 

Sampling Requirements to Demonstrate Process Control in Slaughter Operations 

3/30/2020 

Attachment 2 

MICROBIOLOGICAL TESTING OF RAW POULTRY SUMMARY CHART 

ESTABLISHMENT 

SIZE 

DEFINED AS MINIMUM 

SAMPLING 

EVENT 

FREQUENCY 

MINIMUM 

SAMPLING 

LOCATION 

Very small (VS) Fewer than 10 

employees or annual 

sales of less than $2.5 

million 

At least once 

during each week 

of operation, 

starting June 1 of 

every year. 

If, after 

consecutively 

collecting 13 

weekly samples 

and upon 

demonstrating 

effective process 

control, the 

sampling plan 

may be modified. 

A sample at 

post-chill per 

sampling event 

Very low volume 

(VLV) 

Slaughter no more than 

440,000 chickens, 60,000 

turkeys, 60,000 ducks, 

60,000 geese, 60,000 

guineas, or 60,000 

squabs or a combination 

of all types of poultry not 

exceeding 60,000 turkeys 

and 440,000 birds total 

annually. 

Small 10 – 499 employees 

unless annual sales total 

less than $2.5 million 

Chickens: once 

per 22,000 

carcasses, but at 

a minimum of 

once during each 

week of 

operation. 

Turkeys, ducks,

geese, guineas,

and squabs: 

once per 3,000 

carcasses but at 

a minimum once 

each week of 

operation 

A sample at prechill and a 

sample at postchill locations 

per sampling 

event 

Large 500 or more employees 

Inspection Methods 29-36 
 

 

 

 

 

Sampling Requirements to Demonstrate Process Control in Slaughter Operations 

3/30/2020 

Attachment 3 

MICROBIOLOGICAL TESTING OF RAW SWINE SUMMARY CHART 

ESTABLISHMENT 

SIZE 

DEFINED AS MINIMUM 

SAMPLING 

EVENT 

FREQUENCY 

MINIMUM 

SAMPLING 

LOCATION 

Very low-volume 

(VLV) 

Slaughter no more than 

20,000 swine or a 

combination of swine and 

other livestock not 

exceeding 6,000 cattle 

and 20,000 total of all 

livestock annually. 

At least once 

during each week 

of operation, 

starting June 1 of 

every year, until 

13 consecutive 

samples show 

process control. 

A sample at 

post-chill 

All establishments 

except for very lowvolume 

One sample set 

per 1,000 

carcasses, but at 

a minimum of 

once during each 

week of 

operation. 

A sample at preevisceration and 

a sample at 

post-chill 

locations per 

sampling event 

Inspection Methods 29-37 
 

 

 

 

 

i(JUi 

!(lt4 

1(lt2: 

1(11_0 

5 10 

• 

• •• 

15 20 25 

Sampling Requirements to Demonstrate Process Control in Slaughter Operations 

3/30/2020 

WORKSHOP 

MICROBIAL INDICATOR ORGANISM IN POULTRY SLAUGHTER OPERATIONS 

An establishment that slaughters 22,000 young chickens daily has incorporated its 

written microbiological sampling procedure for analysis of Enterobacteriaceae into its 

Sanitation SOP. You scheduled an Operational SSOP Review and Observation task on 

your task calendar in PHIS for today. 

Scenario 

You review the written sampling procedure used to monitor the effectiveness of the 

procedures the establishment implements to prevent contamination of carcasses and 

parts with enteric pathogens in the QC office. In the sampling procedure, the 

establishment is collecting samples (carcass rinsate) at the required locations and 

frequencies in accordance with 9 CFR 381.65(g)(1)(2). You also review the company’s 

process control chart for the Enterobacteriaceae analyses. The establishment uses a 

moving window of the thirteen most recent tests to evaluate process control. The chart 

shows the following results: 

You conclude that the establishment is maintaining daily records sufficient to document 

the implementation and monitoring of the sampling procedures but note that the SSOP 

corrective action records for the sampling program do not reflect any action taken for 

the last two weeks. Based on the information given in the scenario, answer the following 

questions: 

Inspection Methods 29-38 
 

 

 

Sampling Requirements to Demonstrate Process Control in Slaughter Operations 

3/30/2020 

What do you conclude from the review of the establishment’s process control chart with 

the most recent thirteen tests results? 

Is this a noncompliance? If so, why and cite the noncompliant regulations. 

Inspection Methods 29-39 
 

 

 

 

 

 

 

 

 

 

 

 

Sampling Requirements to Demonstrate Process Control in Slaughter Operations 

3/30/2020 

GENERIC E. COLI TESTING 

General Instructions 

Work through this workshop with at least one partner. 

You have scheduled a generic E. coli verification task on your task calendar in PHIS for 

establishment number M44927 for today. You review the establishment’s generic E. coli 

written procedure and records, as well as the regulations. 

Generic E. coli verification task 

The establishment collects 310.25(a) Observe sample collection and review 

samples from the type of procedures and records. 

livestock it slaughters in FSIS 

greatest numbers; selects Directive Make determinations about compliance 

carcasses randomly; selects 5000.1 with regulatory requirements. 

carcass samples at required 

location in process, and by Document failure(s) to comply with 

procedure specified in regulatory requirements on NR and, 

regulation. when appropriate, take other actions 

consistent with applicable directive(s). 

Scenario 

From the random sample collection times provided to you by Irene Bossley, the QC 

technician, at the beginning of the shift, you decide to observe the second E. coli 

sample collection of the day. You observe the technician putting on sterile gloves and 

randomly collecting one untrimmed half carcass in the cooler. Following the procedure, 

she changes sterile gloves, aseptically sponges’ three sites (the flank, brisket and rump) 

of the selected beef carcass, following the guidelines for proper handling of the sponge. 

You follow her to the in-house microbiology laboratory where a qualified microbiology 

technician is waiting. 

You discuss the testing procedure used in the on-site lab with the lab technician. She 

tells you that the analysis is completed using a test method she found in a peerreviewed microbiology journal two years ago. She says she has memorized the 

technique and does not need to refer to the instructions in the article as she analyzes 

the sample. She does have a copy of the E. coli test procedure in her files and shares it 

with you. 

Finally, you check the company’s process control charts. There is a moving window of 

the thirteen most recent tests. 

Inspection Methods 29-40 
 

 

 

 

 

 

 

Sampling Requirements to Demonstrate Process Control in Slaughter Operations 

3/30/2020 

OPEN BEEF, Inc. 

M44927 

8305 Hawthorne Way

Petaluma, CA 

E. COLI SPECIMEN COLLECTION PROCEDURE 

This is a one-shift establishment that slaughters heifers and steers. Each day of 

operation the Quality Control Manager, or his designee, will collect a half carcass in 

the cooler for each 300 carcasses slaughtered. When selecting a carcass in the 

cooler at the random time, the QC Manager, or his designee, will walk up to the 

selection point and count five half carcasses. He will then select the sixth half 

carcass. 

Open Beef’s average daily production volume is a combination of 500 heifers and 

steers. Based on this volume, one random sample will be taken during the shift the 

first day of sampling. Two samples are then taken for two days in a row. Then the 

three-day cycle begins again. This method is used to take into account the extra 

carcasses produced each day. 

Before the beginning operations, the QC Manager, or his designee, will use a random 

selection computer program to select the time samples on the shift will be collected. If 

a random time occurs during a scheduled company break, it will be discarded. Only 

times within the hours of actual operation will be chosen. These times will be made 

available to FSIS personnel before operations begin. 

Aseptic sampling technique will be used to ensure sample integrity. The sponge 

method, as outlined in the “Guidelines for E. coli Testing for Process Control 

Verification in Cattle and Swine Slaughter Establishments” will be followed to ensure 

sample integrity. Samples will be taken to our own microbiology laboratory for 

immediate analysis using an AOAC Official Testing Method. In the event our 

laboratory cannot conduct E. coli tests, the QC Manager, or his designee, will 

immediately refrigerate the sample. At the end of the shift, the refrigerated samples 

will be sent via overnight Federal Express to the Always Accurate Microbiology 

Laboratory in Rough and Ready, CA, for immediate analysis. 

Lester Moore, Plant Manager January 27, 2011 

WORKSHOP EXAMPLE ONLY – DO NOT DUPLICATE 

Inspection Methods 29-41 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

... 

I I I I I I I I 

Sampling Requirements to Demonstrate Process Control in Slaughter Operations 

3/30/2020 

CFU/cm2 

E. COLI RESULTS CHART 

m, marginal = 0 CFU/cm2 M, unacceptable = 100 CFU/cm2 

130 

120 

M = 100 

90 

m = 0 

80 

70 

60 

50 

40 

30 

20 

10 

5 

0 ▀

▀

▀

▀

▀ ▀

▀ ▀

▀ ▀

▀ ▀

▀

▀

▀ ▀

▀ ▀

▀

▀

▀ ▀ ▀

2012 1-9 1-10 1-11 1-12 1-13 1-16 1-17 1-18 1-19 1-20 1-23 1-24 1-25 1-26 1-27 1-30 1-31 

Test Number 

WORKSHOP EXAMPLE ONLY – DO NOT DUPLICATE 

Based on the information given in the scenario answer the following question: 

Is the establishment in compliance with the generic E. coli testing requirements? Why? 

If not, describe the noncompliance. Give the regulatory citation. 

Inspection Methods 29-42 
 

 

 

 

 

 

 

 

 

 

 

 

Sampling Requirements to Demonstrate Process Control in Slaughter Operations 

3/30/2020 

Regulations 

Livestock (other than swine)

310.25 Contamination with microorganisms; process control verification criteria 

and testing; pathogen reduction standards. 

(a) Criteria for verifying process control; E. coli testing. 

(1) Each official establishment that slaughters livestock must test for Escherichia coli 

Biotype 1 (E. coli) Establishments that slaughter more than one type of livestock or both 

livestock and poultry, shall test the type of livestock or poultry slaughtered in the 

greatest number. The establishment shall: 

(i) Collect samples in accordance with the sampling techniques, methodology, and 

frequency requirements in paragraph (a)(2) of this section; 

(ii) Obtain analytic results in accordance with paragraph (a)(3) of this section; and 

(iii) Maintain records of such analytic results in accordance with paragraph (a)(4) of 

this section. 

(2) Sampling requirements. 

(i) Written procedures. Each establishment shall prepare written specimen collection 

procedures which shall identify employees designated to collect samples, and shall 

address location(s) of sampling, how sampling randomness is achieved, and handling of 

the sample to ensure sample integrity. The written procedure shall be made available to 

FSIS upon request. 

(ii) Sample collection. The establishment must collect samples from all chilled 

livestock carcasses, except those boned before chilling (hot-boned), which must be 

sampled after the final wash. Samples must be collected in the following manner; 

(A) For cattle, establishments must sponge or excise tissue from the flank, 

brisket and rump, except for hide-on calves, in which case establishments must 

take samples by sponging from inside the flank, inside the brisket, and inside the 

rump. 

(B) For sheep, goat, horse, mule, or other equine carcasses, establishments 

must sponge from the flank, brisket and rump, except for hide-on carcasses, in 

which case establishments must take samples by sponging from inside the flank, 

inside the brisket, and inside the rump. 

(iii) Sampling frequency. Slaughter establishments, except very low volume 

establishments as defined in paragraph (a)(2)(v) of this section, must take samples at a 

frequency proportional to the volume of production at the following rates: 

(A) Cattle, sheep, goats, horses, mules, and other equines: 1 test per 300 

carcasses, but, a minimum of one sample during each week of operation. 

(iv) Sampling frequency alternatives. An establishment operating under a validated 

HACCP plan in accordance with §417.2(b) of this chapter may substitute an alternative 

frequency for the frequency of sampling required under paragraph (a)(2)(iii) of this 

section if, 

Inspection Methods 29-43 
 

 

 

 

 

 

 

 

---------------------------------------------------------------------------

---------------------------------------------------------------------------

Sampling Requirements to Demonstrate Process Control in Slaughter Operations 

3/30/2020 

(A) The alternative is an integral part of the establishment's verification 

procedures for its HACCP plan and, 

(B) FSIS does not determine, and notify the establishment in writing, that the 

alternative frequency is inadequate to verify the effectiveness of the 

establishment's processing controls. 

(v) Sampling in very low volume establishments. (A) Very low volume establishments 

annually slaughter no more than 6,000 cattle, 6,000 sheep, 6,000 goats, 6,000 horses, 

mules or other equines, or a combination of livestock not exceeding 6,000 cattle and 

20,000 total of all livestock. Very low volume establishments that collect samples by 

sponging shall collect at least one sample per week, starting the first full week of 

operation after June 1 of each year, and continue sampling at a minimum of once each 

week the establishment operates until June 1 of the following year or until 13 samples 

have been collected, whichever comes first. Very low volume establishments collecting 

samples by excising tissue from carcasses shall collect one sample per week, starting 

the first full week of operation after June 1 of each year, and continue sampling at a 

minimum of once each week the establishment operates until one series of 13 tests 

meets the criteria set forth in paragraph (a)(5)(i) of this section. 

(B) Upon the establishment's meeting requirements of paragraph (a)(2)(v)(A) of 

this section, weekly sampling and testing is optional, unless changes are made in 

establishment facilities, equipment, personnel or procedures that may affect the 

adequacy of existing process control measures, as determined by the 

establishment or FSIS. FSIS determinations that changes have been made 

requiring resumption of weekly testing shall be provided to the establishment in 

writing. 

(3) Analysis of samples. Laboratories may use any quantitative method for analysis of 

E. coli that is approved as an AOAC Official Method of the AOAC International (formerly 

the Association of Official Analytical Chemists)2 or approved and published by a 

scientific body and based on the results of a collaborative trial conducted in accordance 

with an internationally recognized protocol on collaborative trials and compared against 

the three tube Most Probable Number (MPN) method and agreeing with the 95 percent 

upper and lower confidence limit of the appropriate MPN index. 

\2\ A copy of the current edition/revision of the ``Official Methods of AOAC 

International,'' 16th edition, 3rd revision, 1997, is on file with the Director, Office of the 

Federal Register, and may be purchased from the Association of Official Analytical 

Chemists International, Inc., 481 North Frederick Ave., Suite 500, Gaithersburg, MD 

20877-2417. 

(4) Recording of test results. The establishment shall maintain accurate records of all 

test results, in terms of CFU/cm2 of surface area sponged or excised. Results shall be 

recorded onto a process control chart or table showing at least the most recent 13 test 

results, by type of livestock slaughtered. Records shall be retained at the establishment 

for a period of 12 months and shall be made available to FSIS upon request. 

Inspection Methods 29-44 
 

 

 

 

 

 

 

 

 

 

Sampling Requirements to Demonstrate Process Control in Slaughter Operations 

3/30/2020 

(5) Criteria for evaluation of test results. 

(i) An establishment excising samples from carcasses is operating within the criteria 

when the most recent E. coli test result does not exceed the upper limit (M), and the 

number of samples, if any, testing positive at levels above (m) is three or fewer out of 

the most recent 13 samples (n) taken, as follows: 

Table 1 to paragraph (A)(5) — Evaluation of E. coli Test Results 

Type of 

livestock 

Lower limit of 

marginal 

range 

(m) 

Upper limit of 

marginal range 

(M) 

Number of 

sample 

tested 

(n) 

Maximum 

number 

permitted in 

marginal range 

(c) 

Cattle............ Negativea 100 CFU/ cm2 13 3 a Negative is defined by the sensitivity of the method used in the baseline study with a 

limit of sensitivity of at least 5 cfu/cm2 carcass surface area. 

(ii) Establishments sponging carcasses shall evaluate E. coli test results using 

statistical process control techniques. 

(6) Failure to meet criteria. Test results that do not meet the criteria described in 

paragraph (a)(5) of this section are an indication that the establishment may not be 

maintaining process controls sufficient to prevent fecal contamination. FSIS shall take 

further action as appropriate to ensure that all applicable provisions of the law are being 

met. 

(7) Failure to test and record. Inspection shall be suspended in accordance with rules of 

practice that will be adopted for such proceedings upon a finding by FSIS that one or 

more provisions of paragraphs (a) (1)-(4) of this section have not been complied with 

and written notice of same has been provided to the establishment. 

Swine 

Sec. 310.18 Contamination of carcasses, organs, or other parts. 

(c) Official swine slaughter establishments must develop, implement, and maintain 

written procedures to prevent contamination of carcasses and parts by enteric 

pathogens, and visible fecal material, ingesta, and milk contamination throughout the 

entire slaughter and dressing operation. Establishments must incorporate these 

procedures into their HACCP plans, or sanitation SOPs, or other prerequisite programs. 

These procedures must include sampling and analysis for microbial organisms in 

accordance with the sampling location and frequency requirements in paragraphs (c)(1) 

and (2) of this section to monitor their ability to maintain process control. 

(1) Sampling locations. Official swine slaughter establishments, except for very lowvolume establishments, must collect and analyze carcass samples for microbial 

Inspection Methods 29-45 
 

 

 

 

 

 

 

Sampling Requirements to Demonstrate Process Control in Slaughter Operations 

3/30/2020 

organisms at the pre- evisceration and post-chill points in the process. Establishments 

that slaughter more than one type of livestock must test the type of livestock 

slaughtered in the greatest number. Establishments that bone their products before 

chilling (i.e., hot-boned products) must collect and analyze samples at the preevisceration point in the process and after the final wash instead of at post-chill. Very 

low- volume establishments must collect and analyze samples for microbial organisms 

at the post-chill point in the process. All swine establishments must sponge or excise 

tissue from the ham, belly, and jowl areas. 

(i) Very low-volume establishments annually slaughter no more than 20,000 swine, or 

a combination of swine and other livestock not exceeding 6,000 cattle and 20,000 total 

of all livestock. 

(ii) [Reserved] 

(2) Sampling frequency. Establishments, except for very low- volume establishments as 

defined in paragraph (c)(1)(i) of this section, must collect and analyze samples at a 

frequency proportional to the establishment’s volume of production at the following 

rates: 

(i) Establishments, except for very low-volume establishments as defined in 

paragraph (c)(1)(i) of this section, must collect and analyze samples at a frequency of 

once per 1,000 carcasses, but a minimum of once during each week of operation. 

(ii) Very low-volume establishments as defined in paragraph (c)(1)(i) of this section 

must collect and analyze samples at least once during each week of operation starting 

June 1 of every year. If, after consecutively collecting 13 weekly samples, very lowvolume establishments can demonstrate that they are effectively maintaining process 

control, they may modify their sampling plans. 

(iii) Establishments must maintain accurate records of all test results and retain these 

records as provided in paragraph (d) of this section. 

(d) Official swine slaughter establishments must maintain daily records sufficient to 

document the implementation and monitoring of the procedures required under this 

section. Records required by this section may be maintained on computers if the 

establishment implements appropriate controls to ensure the integrity of the electronic 

data. Records required by this section must be maintained for at least one year and 

must be accessible to FSIS. 

Poultry

Sec. 381.65 Operations and procedures, generally 

(g) Procedures for controlling contamination throughout the slaughter and dressing 

operation. Official poultry slaughter establishments must develop, implement, and 

maintain written procedures to prevent contamination of carcasses and parts by enteric 

pathogens and fecal contamination throughout the entire slaughter and dressing 

operation. Establishments must incorporate these procedures into their HACCP plans, 

Inspection Methods 29-46 
 

 

Sampling Requirements to Demonstrate Process Control in Slaughter Operations 

3/30/2020 

or sanitation SOPs, or other prerequisite programs. At a minimum, these procedures 

must include sampling and analysis for microbial organisms in accordance with the 

sampling location and frequency requirements in paragraphs (g)(1) and (2) of this 

section to monitor their ability to maintain process control. 

(1) Sampling locations. Establishments, except for very small establishments 

operating under Traditional Inspection or very low volume establishments operating 

under Traditional Inspection must collect and analyze samples for microbial organisms 

at the pre-chill and post-chill points in the process. Very small establishments operating 

under Traditional Inspection and very low volume establishments operating under 

Traditional Inspection must collect and analyze samples for microbial organisms at the 

post-chill point in the process. 

(i) Very small establishments are establishments with fewer than 10 employees or 

annual sales of less than $2.5 million. 

(ii) Very low volume establishments annually slaughter no more than 440,000 

chickens, 60,000 turkeys, 60,000 ducks, 60,000 geese, 60,000 guineas, or 60,000 

squabs. 

(2) Sampling frequency. (i) Establishments, except for very low volume establishments 

as defined in paragraph (g)(1)(ii) of this section, must, at a minimum, collect and 

analyze samples at a frequency proportional to the establishment’s volume of 

production at the following rates: 

(A) Chickens. Once per 22,000 carcasses, but a minimum of once during each week 

of operation. 

(B) Turkeys, ducks, geese, guineas, and squabs. Once per 3,000 carcasses, but at a 

minimum once each week of operation. 

(ii) Very low volume establishments as defined in paragraph (g)(1)(ii) of this section 

must collect and analyze samples at least once during each week of operation starting 

June 1 of every year. If, after consecutively collecting 13 weekly samples, a very low 

volume establishment can demonstrate that it is effectively maintaining process control, 

it may modify its sampling plan. 

(iii) Establishments must sample at a frequency that is adequate to monitor their ability 

to maintain process control for enteric pathogens. Establishments must maintain 

accurate records of all test results and retain these records as provided in paragraph (h) 

of this section. 

(h) Recordkeeping requirements. Official poultry slaughter establishments must 

maintain daily records sufficient to document the implementation and monitoring of the 

procedures required under paragraph (g) of this section. Records required by this 

section may be maintained on computers if the establishment implements appropriate 

controls to ensure the integrity of the electronic data. Records required by this section 

must be maintained for at least one year and must be accessible to FSIS. 

Inspection Methods 29-47 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sampling Requirements to Demonstrate Process Control in Slaughter Operations 

3/30/2020 

Ratites 

Sec. 381.94 Contamination with Microorganisms; process control verification 

criteria and testing; pathogen reduction standards for establishments that 

slaughter ratites 

(a) Criteria for verifying process control; E. coli testing. (1) Each official establishment 

that slaughters ratites shall test for Escherichia coli Biotype I (E. coli). Establishments 

that slaughter ratites and livestock, shall test the type of ratites or livestock slaughtered 

in the greatest number. The establishment shall: (i) Collect samples in accordance with 

the sampling techniques, methodology, and frequency requirements in paragraph (a)(2) 

of this section; 

(ii) Obtain analytic results in accordance with paragraph (a)(3) of this section; and 

(iii) Maintain records of such analytic results in accordance with paragraph (a)(4) of 

this section. 

(2) Sampling requirements. (i) Written procedures. Each establishment that 

slaughters ratites shall prepare written specimen collection procedures which shall 

identify employees designated to collect samples, and shall address location(s) of 

sampling, how sampling randomness is achieved, and handling of the sample to ensure 

sample integrity. The written procedure shall be made available to FSIS upon request. 

(ii) Sample collection. The establishment must collect samples from whole ratites at 

the end of the chilling process. Samples from ratites may be collected by sponging the 

carcass on the back and thigh or samples can be collected by rinsing the whole carcass 

in an amount of buffer appropriate for that type of bird. 

(iii) Sampling frequency. Establishments that slaughter ratites, except very low 

volume ratite establishments as defined in paragraph (a)(2)(v) of this section, must take 

samples at a frequency proportional to the establishment's volume of production at the 

following rate: 1 sample per 3,000 carcasses, but at a minimum one sample each week 

of operation. 

(iv) Sampling frequency alternatives. An establishment operating under a validated 

HACCP plan in accordance with Sec. 417.2(b) of this chapter may substitute an 

alternative frequency for the frequency of sampling required under paragraph (a)(2)(iii) 

of this section if, 

(A) The alternative is an integral part of the establishment's verification procedures for 

its HACCP plan and, 

(B) FSIS does not determine, and notify the establishment in writing, that the 

alternative frequency is inadequate to verify the effectiveness of the establishment's 

processing controls. 

(v) Sampling in very low volume ratite establishments. (A) Very low volume ratite 

establishments annually slaughter no more than 6,000 ratites. Very low volume ratite 

establishments that slaughter ratites in the largest number must collect at least one 

sample during each week of operation after June 1 of each year, and continue sampling 

at a minimum of once each week the establishment operates until June of the following 

year or until 13 samples have been collected, whichever comes first. 

(B) Upon the establishment's meeting the requirements of paragraph (a)(2)(v)(A) of 

this section, weekly sampling and testing is optional, unless changes are made in 

establishment facilities, equipment, personnel or procedures that may affect the 

Inspection Methods 29-48 
 

 

 

 

 

 

Sampling Requirements to Demonstrate Process Control in Slaughter Operations 

3/30/2020 

adequacy of existing process control measures, as determined by the establishment or 

by FSIS. FSIS determinations that changes have been made requiring resumption of 

weekly testing shall be provided to the establishment in writing. 

(3) Analysis of samples. Laboratories may use any quantitative method for analysis of 

E. coli that is approved as an AOAC Official Method of the AOAC International (formerly 

the Association of Official Analytical Chemists) or approved and published by a scientific 

body and based on the results of a collaborative trial conducted in accordance with an 

internationally recognized protocol on collaborative trials and compared against the 

three tube Most Probable Number (MPN) method and agreeing with the 95 percent 

upper and lower confidence limit of the appropriate MPN index. 

(4) Recording of test results. The establishment shall maintain accurate records of all 

test results, in terms of colony forming units (CFU)/ml of rinse fluid. Results shall be 

recorded onto a process control chart or table showing at least the most recent 13 test 

results. Records shall be retained at the establishment for a period of 12 months and 

shall be made available to FSIS upon request. 

(5) Establishments shall evaluate E. coli test results using statistical process control 

techniques. 

(6) Failure to meet criteria. Test results that do not meet the criteria described in 

paragraph (a)(5) of this section are an indication that the establishment may not be 

maintaining process controls sufficient to prevent fecal contamination. FSIS shall take 

further action as appropriate to ensure that all applicable provisions of the law are being 

met. 

(7) Failure to test and record. Inspection will be suspended in accordance with rules 

of practice that will be adopted for such proceeding, upon a finding by FSIS that one or 

more provisions of paragraphs (a) (1) through (4) of this section have not been complied 

with and written notice of same has been provided to the establishment. 

Inspection Methods 29-49