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Procedures Manual:

Bovine Spongiform Encephalopathy (BSE)

Ongoing Surveillance Program

November 2020

U.S. Department of Agriculture

Animal and Plant Health Inspection Service

Veterinary Services
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Table of Contents

Introduction .............................................................................................................................................. 4

Purpose ......................................................................................................................................................... 4

Surveillance Plan Overview ......................................................................................................................... 4

Targeted Subpopulations of Cattle for BSE Sampling as Part of Ongoing Surveillance ............................. 5

A. Cattle, of any breed 12 months old and older, exhibiting clinical signs of central nervous system (CNS) 

disease (clinical suspects)......................................................................................................................... 5

B. Cattle of any breed that are 30 months old or older exhibiting certain nonspecific clinical signs ...... 6

Collection Sites............................................................................................................................................. 6

Personal Safety ............................................................................................................................................. 8

Detailed Sampling Procedures ..................................................................................................................... 9

Tools needed............................................................................................................................................. 9

Getting a sample of sufficient quality....................................................................................................... 9

BSE sampling using a brain sampling spoon ........................................................................................... 9

Packaging and Shipping Samples........................................................................................................... 14

Packaging materials................................................................................................................................ 14

Packing and shipping.............................................................................................................................. 15

Rabies-negative samples ........................................................................................................................ 15

Designated Laboratories for BSE Sample Submission .............................................................................. 15

Proper Communication for Submitting Samples........................................................................................ 16

Animal Identification.................................................................................................................................. 17

Reporting of Laboratory Results................................................................................................................ 18

Instructions for Completing the BSE Surveillance Forms ......................................................................... 18

General Instructions................................................................................................................................ 18

Instructions for completing USDA BSE Surveillance Submission Form: VS Form 17-146 ................ 19

Instructions for completing USDA BSE Surveillance Submission Continuation Form: VS Form 17-146 A

................................................................................................................................................................ 21

Instructions for completing USDA BSE Surveillance Data Collection Form: VS Form 17-131.......... 22

Humane Handling of Livestock.................................................................................................................. 27

Transportation and Disposal of Carcasses.................................................................................................. 27

Attachment A: BSE Ongoing Surveillance Targeting Criteria Flowchart ................................................ 29

Attachment B: USDA BSE Surveillance Submission Form (VS Form 17-146)....................................... 31

Attachment C: USDA BSE Surveillance Submission Continuation Form (VS Form 17-146A).............. 33

Attachment D: USDA BSE Surveillance Data Collection Form (VS Form 17-131)................................ 34
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Attachment E: IATA 650 Instructions....................................................................................................... 35

Attachment F: BSE Sample Collection Requirements.............................................................................. 38

Attachment G: BSE VSLS Quick Start Guide .......................................................................................... 40
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Introduction 

This document describes the procedures used to implement the Bovine Spongiform Encephalopathy 

(BSE) Ongoing Surveillance Program. This procedures manual is meant to be dynamic and will be 

updated as necessary. 

Several other Veterinary Services (VS) documents contain policy information that supplements the 

instructions described in this manual: 

• BSE Ongoing Surveillance Plan (July 20, 2006)

• VS Guidance Document 7800.1 “Procedures for Conducting the Bovine Spongiform

Encephalopathy (BSE) Ongoing Surveillance Program”

• BSE Response Plan (version 2020) 

• Evaluation of Bovine Spongiform Encephalopathy National Surveillance Program: An analysis 

of the ongoing surveillance program for bovine spongiform encephalopathy in the U.S. cattle 

population (January 2018)

Purpose 

The purpose of this procedures manual is to provide detailed instructions concerning: 

• The targeting criteria for cattle to be sampled as part of this surveillance program; 

• Personal safety guidelines; 

• Sample collection; 

• Data collection; 

• Shipping samples to a laboratory; 

• Communication protocols; and 

• Disposal of the carcass and offal. 

Surveillance Plan Overview 

The BSE Ongoing Surveillance Program, VS’ long-term BSE surveillance program, was implemented in 

August 2006 to follow the Enhanced Surveillance Program in which nearly three quarters of a million 

cattle were tested for BSE. Analysis of data from the Enhanced Surveillance Program revealed that the 

United States is free from BSE with a very low threshold for detection. The principal purposes of 

ongoing surveillance for BSE are to continue to monitor and assess changes to the BSE status of U.S. 

cattle and to provide mechanisms for early detection of BSE. The Ongoing Surveillance Plan is 

designed to exceed World Organization for Animal Health (OIE) Type B surveillance guidelines that 

require adequate surveillance samples to detect one case of BSE per 50,000 adult cattle with 95 percent 
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confidence. The Ongoing Surveillance Plan has adopted a more sensitive design prevalence to meet the 

goal of being able to detect one case of BSE per 1,000,000 adult cattle with 95 percent confidence. 

Targeted Subpopulations of Cattle for BSE Sampling as Part of Ongoing Surveillance

The targeted subpopulations for the BSE Ongoing Surveillance Program include cattle (Bos taurus and 

Bos indicus) from the following surveillance streams, as outlined in the OIE Terrestrial Animal Health 

Code: clinical suspects, casualty (emergency) slaughter, and fallen stock1

. BSE samples are 

assigned to the appropriate OIE surveillance stream based on the submission reason, clinical history 

and/or signs, sample source, and condemnation code, as detailed in the BSE Ongoing Surveillance Plan. 

The targeted subpopulations for BSE sample collection (Attachment A: BSE Ongoing Surveillance 

Targeting Criteria Flowchart), as described in the BSE Ongoing Surveillance Plan, are outlined 

below. 

A. Cattle, of any breed 12 months old and older, exhibiting clinical signs of central nervous 

system (CNS) disease (clinical suspects)

Cattle exhibiting clinical signs of CNS disease are highly valuable with regard to BSE surveillance 

activities. Cattle in this category are a priority for surveillance and every effort should be made to 

sample these cattle for BSE, regardless of the site/location where these animals are identified. 

Cattle that are less than 12 months old and thus exempted from testing are defined in this document by 

dentition as “all teeth that have erupted, both incisors and cheek teeth, are deciduous and none is loose.”

Cattle 12 months old and older highly suspicious for BSE, as described in VS Guidance Document 

7800.1 (or current version)

• Cattle with CNS signs - displaying progressive behavioral changes that cannot be attributed to 

infectious illness (including but not limited to apprehension; nervousness; excitability;

aggression toward other cattle or humans; head shyness; hypermetria; persistent kicking when 

milked; difficulty in rising; excessive nose scratching; changes in herd hierarchical status;

hesitation at doors, gates, and barriers; or reluctance to cross concrete or other “slippery” 

surfaces). These include cattle:

o On the farm with a history of or exhibiting clinical signs of CNS disease;

o Classified as rabies suspects (including rabies-negative cases from State or local public 

health laboratories); and

o Condemned on antemortem inspection for CNS signs or rabies. 

1 Clinical suspects refer to cattle displaying behavioral or clinical signs consistent with BSE. Casualty (Emergency) slaughter 

refers to animals that are non-ambulatory, recumbent, unable to rise or walk without assistance, or “downers.” Fallen stock 

refers to animals that are found dead or killed on-farm or during transport or at slaughter. 
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• Cattle with signs consistent with CNS signs - cattle affected by illnesses that are refractory to 

treatment (including anorexia, loss of condition in spite of good appetite, pneumonia, and 

decreased milk yield) and are displaying progressive behavioral changes (including 

apprehension, nervousness, excitability, aggression, head shyness, hypermetria, kicking when 

milked, difficulty in rising, excessive nose scratching, or hesitation at gates/barriers).

B. Cattle of any breed that are 30 months old or older exhibiting certain nonspecific clinical signs 

Certain cattle, other than those described in Section A above, are also valuable to targeted BSE 

surveillance efforts; however, samples should only be obtained from cattle 30 months old or older (as 

evidenced by the eruption of at least one of the second set of permanent incisors). Categories of these 

targeted cattle include: 

• Cattle condemned on antemortem at slaughter for any reason other than CNS signs or rabies. 

• Non-ambulatory cattle that cannot rise from a recumbent position (i.e., disabled and/or downer) 

or that cannot walk including, but not limited to, those with broken appendages, severed tendons 

or ligaments, nerve paralysis, fractured vertebral columns, or metabolic conditions. 

• Ambulatory cattle at slaughter exhibiting other clinical signs that may be associated with BSE: 

o Cattle that are severely weakened though they may be able to stand and walk for brief 

periods of time; and 

o Cattle that were euthanized, or that died as a result of moribund conditions, infectious 

diseases, emaciation, or injuries. 

• Cattle found dead. 

o A clinical history should be obtained for these animals whenever possible. 

o Samples should not be collected from dead cattle with known cause of death and/or 

history that do not meet the criteria for targeted subpopulations (e.g., acute deaths from 

lightning strike or other acts of nature, fire, dystocia, hypothermia, trauma, etc.). 

Collection Sites 

Cattle in the targeted subpopulations described above may present at various locations. Thus, sample 

collection sites for the BSE Ongoing Surveillance Program include farms, public health and animal 

diagnostic laboratories, and rendering, 3D/4D, and slaughter establishments. 

• Slaughter Establishments. Cattle 12 months old and older condemned for CNS signs or 

rabies upon antemortem inspection will be sampled for BSE as described in Section A above. 

Additionally, cattle 30 months old or older that are condemned at antemortem inspection by the 
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Food Safety and Inspection Service (FSIS) are eligible for sample collection at designated offsite 

sample collection facilities (facilities contracted by VS to collect samples from cattle condemned 

at antemortem inspection). Most samples from cattle condemned on antemortem inspection will 

contribute to the second most valuable surveillance stream (i.e., casualty slaughter).

• Rendering or 3D/4D facilities. A limited number of samples will be collected from cattle 30

months old and older presented to rendering or 3D/4D facilities. Sample collection at these sites 

should only occur according to provisions of competitive contracts awarded by VS. These sites 

typically include cattle from a wide variety of sources, and samples collected at these sites will 

represent the “fallen stock” surveillance stream. Note: For cattle sampled at these locations, the 

collection of a clinical history is highly desirable. 

 

• On-Farm. Samples from cattle 12 months old and older may be collected by accredited 

veterinarians, Federal or State employees (including animal health technicians and/or 

contractors), or VS-approved dead stock haulers. Under VS Area Office oversight, sample 

collectors with other qualifications may be enlisted when resources preclude the participation of 

aforementioned sample collectors in a given area. Such samples are usually accompanied by 

significant historical information pertaining to clinical signs, and are generally of high value to 

surveillance. 

• Public Health Laboratories. All samples from cattle 12 months old and older that are rabies 

suspects and test negative for rabies will be submitted to a designated laboratory (Table 1) for 

BSE testing by public health laboratory personnel. All samples derived from this data source can 

be characterized as clinically suspicious for BSE and thus are of high value for surveillance.

• Veterinary Diagnostic Laboratories. Cattle 12 months old and older that are submitted for 

necropsy, or fresh whole brainstems submitted for ancillary diagnostics to veterinary diagnostic 

laboratories, including laboratories other than VS BSE contract laboratories, should be sampled 

by laboratory personnel. Such samples are usually accompanied by significant historical 

information pertaining to clinical signs, and thus are of high value to surveillance. 

• Facilities contracted by VS to collect samples from cattle condemned at antemortem 

inspection (i.e., offsite collection facilities). Samples derived from cattle 12 months old and 

older presented for slaughter and condemned at antemortem inspection by FSIS may be collected 

by Animal and Plant Health Inspection Service (APHIS) employees (including animal health 

technicians and/or contractors) at an APHIS-approved, offsite collection facility. Under these 

circumstances, communication of animal identification information (including FSIS 

condemnation tag number, i.e., the “Z tag”), clinical history, and condemnation codes from the 

slaughter facility to the contracted facility is imperative. 
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Personal Safety 

If BSE is transmissible to humans in the occupational setting, the most likely routes would be through 

contact with infective tissues through wounds or open lesions on the skin, contact with mucous 

membranes (eyes and mouth), or in exceptional cases, by swallowing. Transmission by the airborne 

route (i.e., by the inhalation of infectious airborne particles) is considered to be the least likely route of 

exposure. The only tissues that have shown infectivity in naturally infected cattle are the brain, retina, 

spinal cord, nictitating membrane, and a limited number of other tissues such as peripheral nerves. In 

experimentally (orally) infected cattle, the distal ileum, dorsal root ganglia, trigeminal ganglia, palatine

tonsil, and a limited number of other tissues has also shown infectivity, the majority of these at end stage 

disease. 

Because rabies, listeriosis, and other possible zoonotic diseases must be included in the differential 

diagnosis, brain and spinal cord collection from cattle with CNS clinical signs should be done carefully. 

The following precautions are generally applicable: 

• Adhere to safe working practices and take extra precautions to avoid or minimize the use of tools 

and equipment likely to cause cuts, abrasions, or puncture wounds; 

• Where use of such equipment is unavoidable, wear suitable protective clothing, which includes 

disposable coveralls, aprons, heavy gloves, and boots; 

• Cover existing cuts, abrasions, and skin lesions on exposed skin with waterproof dressings; 

• Use face protection to protect the mucous membranes of the eye, nose, and mouth from exposure 

to infective droplets or tissue fragments; 

• Take steps to avoid the creation of aerosols and dusts when engaged in activities such as sawing 

through the skull bones;

• Wash hands and exposed skin before eating, drinking, smoking, taking medication, using the 

telephone, or going to the toilet; and 

• Wash and disinfect protective clothing and instruments thoroughly after use. 

Additional guidance for APHIS VS employees concerning personal safety is available in the APHIS 

Safety, Health, and Wellness Manual

(https://www.aphis.usda.gov/aphis/resources/manualsandguidelines/ct_manuals_guidelines_main).
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Detailed Sampling Procedures 

Tools needed 

Knife and scissors (Figure 1) 

Brain sampling spoon or other suitable device

(Figure 1) 

Forceps (Figure 1) 

Screw-top plastic tubes (50ml) (Figure 2) 

Fine point permanent marker 

Ball-point pen 

Pan or bucket for disinfecting instruments 

and rinsing gloved hands 

Bleach (disinfectant) 

Paper towels 

Trash bags 

Supply of BSE mailers including frozen cold packs

May need scabbard, a steel, and personal 

protective equipment

Getting a sample of sufficient quality 

Only samples collected from animals in the targeted subpopulation (as described above) and with a valid 

test result reported are considered to be valid samples in the BSE Ongoing Surveillance Program. “Not 

detected/Not obex” or “Not tested” are not considered valid test results in the context of the BSE

Ongoing Surveillance Program. In order for a sample to be tested, the appropriate brainstem sample, 

including obex, must be submitted with little contamination or postmortem decomposition. Samples 

that are taken from the wrong location or that are significantly autolyzed are not testable and should not 

be submitted. However, samples should be collected and submitted from all cattle condemned by FSIS 

upon antemortem inspection for CNS signs and rabies and all cattle that are highly suspicious for BSE, 

regardless of the apparent tissue quality. 

BSE sampling using a brain sampling spoon

Figure 1: Tools include a knife, scissors, 

brain sampling spoon, and forceps

Figure 2: 50 mL plastic screw-top tube
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Step 1 Place head upright on a head rack or 

barrel, the table edge, or on the ground 

facing down, if no other option is 

feasible. 

Step 1: Positioning the head

Step 2 Grasp the brainstem with forceps, 

using light pressure so that the tissue is 

not damaged. 

Step 2: Grasping the brainstem

Step 3 Cut the dura mater (tough fibrous 

membrane) along the dorsal (top) 

midline to expose the brainstem and 

cut the cranial nerves, present on the 

lateral (sides) aspects of the brainstem, 

to allow for better mobility within the 

foramen and subsequent removal. 

Cut the cranial nerves on the sides. Do 

not cut into the brainstem! Failure to 

sever cranial nerves is a common cause 

of damaged samples. Step 3: Cutting the dura mater and 

the cranial nerves
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Step 4 With forceps and scissors, remove as 

much dura mater as possible. Dura 

mater removal allows better 

visualization and may aid proper 

sample removal. 

Step 4: Removing dura mater to 

visualize the spinal cord

Step 5 With light pressure, use the forceps to 

hold the brainstem against the ventral 

part of the foramen magnum. 

 

Insert the brain sampling spoon 

(inverted) on the dorsal part of the 

brainstem to sever the cerebellar 

attachments to the brainstem. 

 

Remove the spoon. 

Step 5: Severing the cerebellar 

attachments to the rostral brainstem

with a brain sampling spoon

Step 6 With the forceps, lift the brainstem

dorsally and reinsert the brain 

sampling spoon along the ventral 

surface of the spinal cord. 

 

Lower the handle of the spoon, 

elevating the leading edge of the 

spoon, to sever the rostral brainstem. 

Depending on the size of the spoon and 

the animal being sampled, this may 

need to be repeated, directing the 

leading edge laterally left and right. 

 

With constant pressure, gently remove 

the spoon and accompanying brainstem 

through the foramen. If the sample 

Step 6: Severing the rostral brainstem 

with a brain sampling spoon
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does not come freely with the spoon, 

ensure the cranial nerves are 

completely severed along the lateral 

brainstem and repeat steps 5 and 6. 

Forcing removal of the sample can 

result in its mutilation and an invalid 

test result. 

Step 7 Complete the removal of the sample 

from the foramen. 

Gently clean off excess blood. 

Step 7: Removal of the sample

Step 8 Identify the obex area of the brainstem. 

The obex, where the 4th ventricle 

transitions to the central canal of the 

spinal cord, includes the dorsal motor 

nucleus of the vagus (DMNV), which

MUST be collected in order for the 

sample to be counted as a valid sample 

in the BSE Ongoing Surveillance 

Program. The triangle in the figure to 

the right demonstrates the location of 

the DMNV, the pink area on the dorsal 

midline at the obex, and is the portion

examined in the laboratory. The 

arrow identifies the “V” at the level 

of the obex.

Step 8: Identification of the obex
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Step 9 Cut the sample as pictured. The 

middle piece of tissue contains the 

obex and the DMNV.

Remember: THE OBEX IS THE 

DESIRED AREA.

Step 9: Sections of the tissue sample

Step 10 Remember the sample will be FRESH

tissue; NO FORMALIN. Do not 

FREEZE samples unless advised to do 

so by the National Veterinary Services 

Laboratories (NVSL).

Place the obex in the supplied 50 ml 

screw top tube. 

Label the sample tube with: Sample 

number (ex: 1, 2, 3, 4) and Barcode ID 

label. 

Cattle highly suspicious for BSE 

should have their entire brain 

submitted after rabies testing, if 

possible. The entire brainstem can be 

submitted fresh, with the remaining 

brain being split along the midline and 

submitted half fresh and half fixed. 

Contact NVSL for further instructions.

Dispose of un-submitted tissue with 

carcass. 

 

Step 10: Obex placed in screw top tube
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Packaging and Shipping Samples 

It is important that samples are submitted to the testing laboratory as quickly as possible in order to 

minimize time between sample collection and processing. Samples should be shipped on the same day 

as they are collected. Samples should not be saved or batched for shipping at a later date as this 

increases the time between sample collection and carcass identification, including DNA matching. 

Samples should be shipped chilled or refrigerated. Freezing of samples should be kept to a minimum. 

Proper recognition of the brainstem, particularly the obex, is adversely affected by repeated freeze/thaw 

cycles. Lack of identification of the obex, results in a “Not detected/Not obex” diagnosis when tested. 

This means the laboratory did not detect the abnormal prion, but they are not certain that the correct 

region was tested. This is why it is very important to submit intact and recognizable tissues when 

shipping samples for BSE testing. 

Packaging materials 

The National Veterinary Services Laboratories (NVSL) provides all the supplies needed for collection 

and shipping (including an approved shipping box) as a “BSE kit.” These kits include:

• Approved shipping box; 

• Sample tubes (Kits of 5, 10, or 25 tubes are available through NVSL. See hyperlink below.);

• Plastic bag or zip-type bag in which to place sample tubes;

• Barcode ID label; 

• USDA BSE Surveillance Submission Form (Attachment B: VS Form 17-146); 

• USDA BSE Surveillance Submission Form Continuation (Attachment C: VS Form 17-146A); 

• USDA BSE Surveillance Data Collection Form (Attachment D: VS Form 17-131); 

• Absorbent material; 

• Ice packs; 

• Biohazard bags (1 or 2) to comply with the International Air Transport Association (IATA) 

shipping regulations (Attachment E: IATA 650 Instructions); and

• Labels for shipping regulations compliance (e.g., air eligible, IATA statement, UN 3373, Keep 

from heat/freezing, Animal Diagnostic Specimen). 

To request additional “BSE kits” or for further assistance with shipping, you may visit here or contact 

the NVSL shipping department at: 

National Veterinary Services Laboratories 

1920 Dayton Avenue 

Ames, IA 50010 

Phone: (515) 337-7530 
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Fax: (515) 337-7378 

E-mail: NVSLShipping@usda.gov

Packing and shipping 

1. Place labeled sample tubes into the plastic bag with absorbent material; 

2. Place plastic bag into the clear biohazard bag (STP-741) and seal; 

3. Place this bag into the white biohazard bag (STP-740) and seal; 

4. Place the white biohazard bag (STP-740) into your shipping box; 

5. Place frozen ice packs on top of the white biohazard bag (STP-740) and cover with inner 

Styrofoam lid; 

6. Place a copy of the completed USDA BSE Surveillance Submission Form (VS 17-146) and 

USDA BSE Surveillance Data Collection Form (VS 17-131) on top of inner Styrofoam lid; 

7. Seal box; 

8. Apply the overnight shipping label to the box addressed to the appropriate laboratory conducting 

BSE testing (Table 1); 

9. Apply the other required shipping labels to the box; and 

10. Ship by overnight delivery with the Federal contract service. If shipping on a Friday, be sure to 

mark/label box for Saturday delivery. 

Note: Individuals who ship surveillance samples to laboratories are responsible for meeting 

requirements for the shipment of biological substances. At a minimum, these individuals should be 

familiar with the IATA Packing Instructions (Attachment E: IATA 650 Instructions). 

Rabies-negative samples 

Rabies-negative samples should be submitted for BSE testing within 10-14 days of collection. If they 

cannot be submitted within 5-7 days (in refrigerated/chilled conditions), then they can be frozen for 

approximately another week. These timeframes have to do with the condition of the tissues, as the brain 

is quite sensitive to autolysis. Public health labs require the entire fresh brain for rabies testing. Their 

ideal transit time (collection to testing) should be less than 48 hours. (Refrigeration preserves the 

sample for “at least 48 hours.”) The surveillance samples that the labs receive are commonly tested 3-4 

days post collection and generally arrive in acceptable condition, having been chilled nearly that entire 

time.

Designated Laboratories for BSE Sample Submission 

Sample collectors will submit fresh tissue samples to the designated laboratories listed here unless 

otherwise directed by NVSL. 

Table 1: Designated laboratories for BSE sample submission 
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State where sample was collected Designated Laboratory

Alabama, Arizona, Colorado, Florida, Kansas, 

Montana, North Carolina, North Dakota, 

Nebraska, Utah, Wyoming

Colorado State University

Veterinary Diagnostic Laboratory

2450 Gillette Drive

Fort Collins, CO 80526

(970) 297-1281

Arkansas, Connecticut, Delaware, Georgia, 

Illinois, Louisiana, Maine, Maryland,

Massachusetts, , , Missouri, New Hampshire, New 

Jersey, New York, Ohio, Oklahoma, 

Pennsylvania, Rhode Island, South Carolina, 

Tennessee, Vermont Virginia, , West Virginia

Athens Veterinary Diagnostic Laboratory

501 DW Books Drive

University of George

Athens, GA 30602

(706) 542-5568

Idaho and others as requested or redirected USDA, APHIS, VS

National Veterinary Services Laboratories 

(NVSL)

1920 Dayton Avenue

Ames, IA 50010

(515) 337-7526

Alaska, California, Hawaii, Iowa, Kentucky, 

Minnesota, Nevada, New Mexico, , Oregon, 

South Dakota, Washington

Washington Animal Disease Diagnostic 

Laboratory

Bustad Hall, Room 155-N

Pullman, WA 99164

(509) 335-9696

Indiana, Michigan, Mississippi, Texas, Wisconsin Wisconsin Veterinary Diagnostic Laboratory

University of Wisconsin – Madison 

445 Easterday Lane

Madison, WI 53706

(608) 262-5432

Proper Communication for Submitting Samples 

It is essential to have secure and reliable communication among the individuals responsible for sample 

collection at collection locations, the establishment’s management, and NVSL or designated 

laboratories. Communication guidelines are as follows: 

• The sample submitter will notify the appropriate laboratory of incoming samples via fax, 

telephone, e-mail, or any other approved electronic method (including the Veterinary Services 

Laboratory Submission (VSLS) System), unless otherwise instructed by that laboratory. 

• The information to be communicated should include the overnight contract delivery service 

tracking number, the collection site name and address, the unique Referral Number of the 

submission, and the number of samples. 
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• Sample submitters must accurately record all relevant information on the current versions of the 

USDA BSE Surveillance Submission Form (VS 17-146), USDA BSE Surveillance Submission 

Continuation Form (if used), and on each of the USDA BSE Surveillance Data Collection Forms

(VS 17-131). See the section “Instructions for Completing the BSE Surveillance Forms” in this 

Procedures Manual for more details. 

o Enter this information on the electronic version of these forms – either on a tablet PC or 

via the Web-based VSLS System. If electronic entry is unavailable, complete the forms 

by hand and submit for data entry as instructed by the VS Area Office. 

o Print a copy of the completed BSE Surveillance Submission Form, BSE Surveillance 

Submission Continuation Form (if used), and each BSE Surveillance Data Collection 

Form to accompany the samples shipped to the designated laboratory. 

o Prepare four additional copies of these completed forms for further distribution and filing 

(i.e., one retained by the submitter, one for the collection site, one for the VS Area Office, 

and one to be maintained with the animal identification devices). 

o FSIS personnel sampling cattle condemned on antemortem inspection for CNS signs or 

rabies as part of the BSE Ongoing Surveillance Program should also complete the forms 

and either enter data directly into the VSLS System or forward the completed forms to 

the appropriate VS Area Office by fax or e-mail. 

• The sample submitter should verify via the overnight contract delivery service tracking system 

that the submission has been delivered to the designated laboratory. If the sample does not arrive 

as expected, the sample submitter should work with the delivery service to track the location and 

delivery status of the sample. 

Animal Identification 

The collector should maintain the identity of cattle tested as part of the BSE Ongoing Surveillance 

Program throughout the entire sample collection, submission, and testing process. If testing should 

confirm an animal was infected with BSE, the animal’s identification will allow VS personnel to trace 

its origin. 

Record ALL types of animal identification devices, brands, and tattoos associated with the sampled 

animal on the current version of the BSE Surveillance Data Collection Form and enter this information 

in the VSLS – BSE Module. These identification devices may remain intact on the carcass or may be 

collected along with an attached piece of ear tissue (at least dime-sized – 1.8 cm) in a labeled bag and 

maintained until test results are received for the animal. When appropriate, obtain digital pictures or 

drawings of brands and/or collect tissue containing the tattoo. If several animals with no identification 

are sampled from the same premises, an official ID may be applied in order to differentiate carcasses 

after testing. Use the best method to ensure that the carcass can be located if the test result is positive.
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Reporting of Laboratory Results 

When tests are completed and all animals tested in the lot are reported as rapid screening test “Not 

Detected” (i.e., “negative”), the designated laboratories will report rapid screening test results back to 

the sample submitter, the Area Veterinarian in Charge (AVIC), and State Veterinarian. If requested, 

results will also be transmitted to the appropriate management at the collection site. 

The designated laboratories will report any “initial inconclusive” results to NVSL. Contact attempts 

should be made in the following order until contact with NVSL is made: the NVSL Director, the NVSL 

Associate Director, Diagnostic Bacteriology and Pathology Laboratory Director, or Head of Pathology 

Section (designated laboratories will be provided with a phone list along with the standard operating 

procedure for performing BSE testing). No other contacts or reporting of test results from that lot are to 

be made until the repeat testing has been completed. If either of the repeat tests is above the negative 

cut off, then the screening test is considered to be “Inconclusive.” 

If test results for any of the animals in the lot are inconclusive, the testing laboratory’s Director will first 

report the findings to the Director of the NVSL. In addition, the testing laboratory’s Director shall also 

notify the AVIC and State Veterinarian in the State in which the sample was collected and/or the State 

where the carcass is being held pending test results. The information reported shall specify which 

carcass(es) tested inconclusive. The AVIC will report results for the entire lot of samples to the 

submitter only after the VS Deputy Administrator’s Office or District Director has given permission to 

release such information. A decision to hold or dispose of the carcass(es) pending confirmatory testing 

should be made with the concurrence of the AVIC. 

All samples with inconclusive results must be immediately forwarded to the NVSL, with prior 

notification and confirmation of arrival. All confirmatory test results will be transmitted directly to the 

VS Area Office. The AVIC will contact the sample collector and the facility where the sample was 

collected so that carcass disposal can be coordinated and verified. 

Instructions for Completing the BSE Surveillance Forms 

General Instructions 

Subsequent to sample collection, the collector must thoroughly and accurately complete the current 

versions of the USDA BSE Surveillance Forms to ensure proper documentation, data entry, and 

submission of information concerning the samples tested for BSE. This information is necessary to 

assign samples to the appropriate OIE surveillance stream and, thus, determines the epidemiologic value 

of each sample to the BSE Ongoing Surveillance Program. 

The versions of USDA BSE Surveillance Forms currently in use are: 

• USDA BSE Surveillance Submission Form (Attachment B: VS Form 17-146) 
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• USDA BSE Surveillance Submission Continuation Form (Attachment C: VS Form 17-146A)

• USDA BSE Surveillance Data Collection Form (Attachment D: VS Form 17-131)

Examples of the forms are included as Attachment B, Attachment C, and Attachment D of this Manual. 

Copies of the forms are available for downloading at: 

https://www.aphis.usda.gov/aphis/resources/forms/ct_vs_forms

There are two methods for electronically submitting this information: via the BSE VSLS Webpage or 

via tablet PCs at high-volume collection sites. The BSE VSLS Web page is used for entering and 

transmitting data from handwritten USDA BSE Surveillance Submission Forms. This system requires 

an APHIS-assigned log-in and password to enter information about each sample prior to shipping the 

sample to the designated laboratory. After electronically entering and submitting the information, the 

data are then stored with the National Animal Health Laboratory Network in a USDA database. 

A Quick Start Guide for using the Web-based forms is available on the VSLS Website after user log-in. 

The Quick Start Guide is included as Attachment G in this manual. 

If electronic entry is unavailable, handwritten forms are submitted as instructed by the VS Area Office. 

Instructions for completing USDA BSE Surveillance Submission Form: VS Form 17-146 

Note: A separate submission form must be completed for each collector, collection site, and collection 

date combination. 

Collection Site Type: Mark type of location at which the sample(s) was collected. Collection Site 

Types are described on pages 7 and 8 of this manual. If the Collection Site Type does not fit those 

described in this manual, indicate “Other” and describe the site type. If a collector obtains samples from 

more than one site type, a separate submission form must be used for each set of samples from each site 

type. 

Testing Laboratory Use Only (Accession or Identification Number): This section is reserved for the 

testing laboratory use only. Only laboratories should mark in this area. 

BSE Referral Number: The BSE Referral Number must be recorded at the top of each Data Collection 

Form. The number must be a unique identifier for the submission that will not be duplicated in any 

other BSE surveillance submission. The BSE Referral Number is used to associate the BSE 

Surveillance Submission Form to the BSE Surveillance Data Collection Form in the database. 

Manually enter a number that uniquely identifies a lab submission using the following recommended 

format: 

• First two characters indicate the State code: e.g., CO for Colorado, or WA for Washington; 
20

• Next two to three characters are the collector’s initials: First, Middle (if applicable), Last; 

• Next eight characters are the collection date in the MMDDYYYY format: e.g., 04072007 (April 

7, 2007); and 

• The last character is a letter to distinguish between multiple submissions/shipments on the same 

day: A = First, B = Second, etc. (Explained in simple terms, one letter per each shipped box).

Example 1. COSAJ06012006A 

Translates to: Colorado – Steven Allen Jones – June 1, 2006 – first submission of the 

day. 

Example 2. COSAJ06012006B 

Translates to: Colorado – Steven Allen Jones – June 1, 2006 - second submission by 

Steven Allen Jones for that day, either from the same collection site or a different 

collection site. 

Collection Site: Ensure that the National Premises Identification Number (if available for the Collection 

Site) or the FSIS Establishment Number where the sample was collected is entered. Enter all the 

requested data for the Collection Site. If samples are collected from more than one site, a separate 

submission form must be used for each Collection Site. 

Collected By: Enter all the information requested for the person that actually collected the tissue sample 

for submission to the testing laboratory. If the Collector is the same as the Submitter, it is only 

necessary to check the indicated box. 

Submitted By: Enter all the information requested for the person submitting the sample to the 

laboratory. If the samples are being submitted to the NVSL and the Submitter has an NVSL Submitter 

ID, please provide it. 

Sample Information: Enter the total number of samples that are included with the submission in the 

appropriate space. In addition, check whether the preservation method for shipping is “ice pack” or 

“other.” If “other” is checked, enter the preservation method in the “Additional Data” field at the 

bottom of the form. 

BSE Sample ID: Attach a sample ID barcode label for each sample submitted with this form. Barcodes 

are available in the sample kits or can be ordered from NVSL using the following e-mail address: 

nvslshipping@aphis.usda.gov. 

If barcodes are not available at the time of sample collection, the VS Area Office should be contacted so 

that barcodes can be assigned for the submission. The number of barcodes should equal the number of 

samples for the submission. 

Additional Data: Enter any additional information about the submission. 
21

Collection Date: Enter the date the samples were collected. All samples on one form must be collected 

on the same day. (Use the MM/DD/YYYY format.) 

Shipping Date: Enter the date the samples were shipped to the laboratory. (Use the MM/DD/YYYY 

format.) 

Destination Lab: Enter the name of the Laboratory (or the Laboratory ID) where the samples are being 

sent for diagnostic testing. 

Signature of Submitter: Submitter must sign the form. 

Condition: For laboratory use only. 

Distribution: For laboratory use only. 

Received By: For laboratory use only. 

Received Date: For laboratory use only. 

Shipment Tracking No.: Enter the Airbill/Shipment Tracking Number for the package being sent to the 

diagnostic laboratory. 

Instructions for completing USDA BSE Surveillance Submission Continuation Form: VS 

Form 17-146 A 

Note: The paper version of the BSE Surveillance Submission Form has space to indicate the 

identification numbers for 20 animals. If additional animals are sampled, a BSE Surveillance 

Submission Continuation Form listing the unique identification numbers for each additional animal 

should also be submitted. 

BSE Referral Number: Enter the referral number from the BSE Surveillance Submission Form. 

BSE Sample ID: Place the barcode assigned to the sample in this box. 

Additional Data: Enter any additional information about the submission. 

Signature of Submitter: Submitter must sign the form. 
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Instructions for completing USDA BSE Surveillance Data Collection Form: VS Form 17-

131

Note: A separate data collection form must be completed for each sample collected.

Collection Date: Enter the date the sample was collected. This date should match the collection date on 

the BSE Surveillance Submission Form. (Use the MM/DD/YYYY format.) 

BSE Referral Number: Enter the referral number from the BSE Surveillance Submission Form, which 

must accompany this data collection form. 

BSE Sample ID: Place the barcode assigned to the sample in this box. The barcode should correspond 

to the barcode on the accompanying BSE Surveillance Submission Form for that sample. 

Primary Reason for Submission: Use professional judgment to select the one choice that best describes 

the primary reason this sample is being taken. If more than one reason for submission is applicable, 

check the primary reason for submission on the data collection form with the smallest number that 

applies. 

• “Highly suspicious for BSE” if the animal being sampled is demonstrating clinical signs of, or 

has a clinical history consistent with, the definition of “highly suspicious for BSE” as described 

in VS Guidance Document 7800.1. 

• “FSIS, antemortem condemned cattle” for animals that are condemned by FSIS personnel prior 

to slaughter and are sampled at the slaughter plant or at an offsite collection facility. (Note: If 

“FSIS, antemortem condemned cattle” is selected, an FSIS Condemnation Code must also be 

selected at bottom of form.) 

• “Rabies suspect” for all animals that were initially identified for testing for rabies because of 

clinical signs or clinical history as described in VS Guidance Document 7800.1. 

Note: If the animal was condemned by FSIS on antemortem inspection for rabies, the 

primary reason for submission is “FSIS, CNS antemortem condemned cattle.” 

• “CNS signs” if the animal is being sampled because it has central nervous system signs 

consistent with primary CNS disease, but does not meet all the criteria for the “Highly suspicious 

for BSE” category. 

Note: If the animal was condemned by FSIS on antemortem inspection for CNS signs, the 

primary reason for submission is “FSIS, CNS antemortem condemned cattle.” 

• “Non-ambulatory/Disabled/Downer” if the animal is being sampled primarily because it is nonambulatory, periodically or continuously. 

Note: If the animal was condemned by FSIS on antemortem inspection as non-ambulatory, 

the primary reason for submission is “FSIS, antemortem condemned cattle” and a 

condemnation code of “Non-ambulatory 445” is assigned to the animal. 
23

• “Other clinical signs that may be associated with BSE” if the animal is being sampled because it 

has demonstrated clinical signs that may be consistent with BSE, such as emaciation, tetanus 

(tetany), or injuries.

• “Dead” if the available history only indicates that the animal is dead with none of the preceding 

conditions as described above. 

Note: If the animal is dead AND an FSIS antemortem condemned animal, the primary 

reason for submission is “FSIS, antemortem condemned cattle” and a condemnation code of 

“Dead 603” is assigned to the animal.

Individual who determined primary reason and clinical signs: Select (check) one box for the individual 

who determined the primary reason for this submission and the clinical signs for the animal identified on 

this form. If “Other” is selected, describe this individual on the line provided on the form. 

Owner Information: Enter as much information as is known for the premises on which the sampled

animal last resided or was held. At a minimum, the State or country is required. 

Slaughter Site: Mark the box if the slaughter site is the same as the collection site; otherwise, enter all 

the requested information on the slaughter site. Be sure to enter the FSIS Establishment Number or the 

National Premises Identification Number (if available). Leave this section blank for non-slaughter 

animal samples. 

Animal Information: Enter all information as requested. 

Breed (If known): Enter the apparent breed of the animal. If unknown, check either “Beef breed”

or “Dairy breed.”

Primary colors: Enter the animal’s primary color(s). 

Age: Enter the animal’s age as the number of months or the number of years (in whole numbers 

only). For instance, if the animal is 2 1⁄2 years old, enter 30 months. If purebred records or other 

official sources of age are used as the source of the animal’s age, check the “Recorded” box. 

Otherwise, indicate that the age was estimated by checking the “Estimated” box. 

Dentition: Examine the animal’s mouth to determine if at least one of the 2nd incisors has erupted. 

If so, check the “Yes” box. 

Gender: Check the appropriate box: “Male,” “Female,” or “Unknown.”

Neutered: Check the appropriate box: “Yes,” “No,” or “Unknown.”
24

Country of Origin: If it is known that the animal originated from a country other than the United 

States, write the name of the country in the space provided. 

Animal ID Information: In the appropriate boxes, enter ALL types of animal identification devices, 

brands, and tattoos associated with the sampled animal. 

• Be sure to include the FSIS Condemnation Tag (“Z-tag” number) of FSIS-condemned animals. 

• If the animal is branded with either a hot iron brand or freeze brand, describe that brand to the 

best of your ability. For example, write “Circle Bar T” in the appropriate box. 

Detailed instructions for specific types of animal identification devices follow. 

• General: 

o All alpha characters (letters) should be recorded in upper case (capitalized). 

o Substitute an underscore (_) without additional spaces, for each unreadable number/character 

in the identification. 

o If more than two official identifications of the same type are available, enter the additional

information in one of the “Other ID” boxes and label accordingly.

o When entering Animal ID for other than “Official” tags, be sure to indicate the color of the 

Animal ID tag, if applicable. Indicate the color for tags other than “Official” tags, if 

applicable, as follows: 

o Begin the Tag ID with the single capitalized letter indicating the color: 

Capital Letter Color Capital Letter Color

Y yellow R red

G green W white

B blue O orange

L lavender (purple) P pink

T tan (brown) S silver 

K black A gray

o Include an underscore (_) between the color-designating letter and the alphanumeric 

sequence of the tag. 

o Record whatever is written or printed on the tags. 

o For two-colored tags, designate the major color in the database but include both colors on the 

paperwork. Example: A yellow plastic bangle tag with the number 25 is “Y_25.” 

o Each identification field referenced on the electronic forms is limited to 15 alpha/numeric 

characters. If additional space is needed to record additional tag numbers, enter the 

information on Line 10, “Additional Data/Comments” on VS Form 17-131, BSE Data 

Collection Form.
25

• Official USDA Tags: 

o USDA bright metal eartags, brucellosis vaccination eartags, Animal Identification Number 

(AIN) eartags, and bangle eartags (official identification tags have the US shield stamped on 

them): 

Note: Back tags can be used in lieu of Official ID on an animal at slaughter. These tags should be 

recorded along with any other tags including official ID.

• Enter the identification information in the appropriate box. 

• No color indication is required. 

• Enter up to two identifications, one in each of the appropriate boxes. 

• If there are more than two official identifications of the same type, enter the additional 

information in one of the “Other ID” boxes and label accordingly. 

o FSIS Condemnation (“Z”) Tags: 

• Enter the number on the FSIS condemnation tag next to the “Z” prefix in the box. 

• When entering the FSIS condemnation tag electronically, include the “Z” prefix with the 

number. Do not include a hyphen (-) after the “Z.”

• Owner Eartag #: 

o Enter information present on the Owner Eartag in the appropriate box. 

o Indicate the color of the tag using the color code listed above, followed by the tag number. 

• Collection Site Tracking #: 

o Enter the Collection Site Tracking Number in the appropriate box. 

• Slaughter Tracking #: 

o Enter the Slaughter Tracking Number in the appropriate box. 

• Back Tag #: 

o Enter the Back Tag Number in the appropriate box. 

• Microchip #: 

o Enter the Microchip Number in the appropriate box. 

• Other ID Type #: 
26

o Flop Tag: Indicate the color of the tag, if applicable. 

o Brands: Describe the brand, to the extent possible, and indicate the location of the brand on 

the animal. 

Note: The electronic data entry form will only accept 15 characters. If you need more than 

15 characters to describe the brand, indicate on the electronic data entry form to refer to the 

paper form for a complete description of the brand.

o Tattoos: Indicate which ear (R = right, L = left) or other location on the animal was tattooed 

and include all alpha/numeric characters. 

Clinical Signs: Check all that apply. At least one clinical sign must be checked. Be thorough and 

complete. Obtain information directly from on-site farm personnel whenever possible. The subtle and 

nonspecific nature of behavioral changes associated with BSE are best identified by those who handle 

cattle on a daily basis, and on-farm personnel are the best source of this information. Use professional 

judgment in box selection and ensure that contradictory signs are not checked. If “Other” is checked, be 

sure to indicate that clinical sign in the “Comments” section at the bottom of the page. 

For clinical signs checked within the black-bordered box on the left-hand side of the form, check 

whether the condition was progressive and whether the condition did not respond to treatment (“Yes,”

“No,” “Don’t Know”). (If the condition did not respond to treatment, the appropriate response is “Yes;”

if the condition did respond to treatment the appropriate response is “No.”) 

Note: Apprehension, ataxia, and hyperesthesia are the three most typical signs of BSE. These are 

identified in the vast majority of animals that are histopathologically confirmed BSE cases. The key 

is to identify an animal with progressive behavioral changes or conditions that are refractory to 

treatment. Many BSE cases are unresponsive to treatment, showing non-specific conditions such as 

the loss of body condition despite good appetite (weight loss), pneumonia, and decreased milk 

production. 

Table 2: Clinical signs commonly identified in BSE-positive animals 

Apprehension

(Increased fearfulness)

Ataxia

(Lack of coordination)

Hyperesthesia

(Excessive sensitivity to sound and 

touch)

Increased aggressive or

belligerent behavior to other

cows and workers due to

increased fear (head butting,

charging, pawing, stamping on

the ground) 

Shortened strides General muscle and head

tremors

Increased resistance to handling Swaying gait Head shyness and ear shifting

(head bobbing, head tossing)

Increased hesitation at doors,

gates, and barriers (i.e., reluctance

to cross concrete or other

“slippery” surfaces)

Difficulty in negotiating turns Rhythmic and involuntary

movement of the eyeballs

(nystagmus)

Head pressing/rubbing, circling Knuckling, stumbling, slipping or Sensitivity to light
27

falling with subsequent difficulty

in rising

Hyper-excitability or nervousness;

may kick repeatedly during

milking (when did not before)

‘Dog sitting’ Excessive or prolonged episodes

of grooming and licking (nose

licking, nose wrinkling, snorting,

yawning, sneezing, and coughing)

Separation from the herd because

animals have become fearful of

their herd-mates or environment

Hypermetria Bruxism (teeth grinding)

Dullness over prolonged periods Progressive weakness Excessive bellowing (vocalization)

Recumbency/paralysis Sudden jerking movements of

head, limbs, or other body parts

to loud sounds (spontaneous

startle responses)

Abnormal head carriage, droopy

lips or eyelids

FSIS Condemnation Codes: Enter the appropriate antemortem condemnation code. One box must be 

checked if and only if “FSIS, antemortem condemned cattle” has been selected as “Primary Reason for 

Submission” on the form. 

Comments: Include any additional relevant information about the sample in this box. 

Humane Handling of Livestock 

Handling procedures and euthanasia for targeted animals tested as part of the BSE Ongoing Surveillance 

Program must comply with applicable local, State, and Federal laws, regulations, and policies, and local 

and professional standards for humane handling and euthanasia of livestock. 

Transportation and Disposal of Carcasses 

Procedures for the transportation and disposal of animal carcasses, parts of dead animals, offal, and 

animal waste for targeted animals tested as part of the BSE Ongoing Surveillance Program should 

comply with local, State, and Federal laws, regulations, and policies, and local and professional 

standards, as well as the following general guidelines: 

• Report domestic animals known to have died of a dangerous transmissible disease or reportable 

disease to the VS Area Office. 

• Prevent contamination of the environment. 

• Prevent exposure to other living animals and the public, including prohibiting the potential for 

uncooked and/or unprocessed animals, animal parts, or offal to be fed to domestic animals. 

• Complete disposal within 48 hours after receipt of BSE test result. 

All carcasses and offal from sampled animals that are intended for further processing must be 

held until laboratory test results are received. Furthermore, the identity of sampled carcasses and 
28

associated offal must be documented and maintained throughout the entire sample collection, 

submission, and testing process until “Not Detected” (i.e., “negative”) test results are obtained. After 

obtaining “Not Detected” test results, carcasses and offal may be rendered or otherwise processed in 

accordance with applicable Federal laws, regulations, and policies. 

Carcasses and offal from sampled animals that are not intended for further processing may be disposed 

of prior to receiving test results by: 

• Burial in order to facilitate DNA matching of positive samples:

o In an approved landfill, OR

o On the farm of immediate origin, OR

o On a composting pile.

Burial of sampled carcasses is preferred to other disposal methods.

Note: It is necessary to mark the location of carcass disposal in the event it needs to be 

exhumed. For landfill disposal, ensure the landfill management is advised of the marker.

• Rendering at a dedicated facility for non-animal feed use such as biofuel or cement; 

• Incineration by fixed-facility incineration; OR

Note: Fixed-facility incineration takes place in a completely contained environment and 

is highly controlled. Fixed-facility incinerators are typically fueled by diesel, natural 

gas, or propane. Carcasses are reduced to inert ash. 

• Alkaline digestion. 

Hides of sampled animals do not require disposal and do not need to be held while awaiting test results. 

If testing confirms that an animal was infected with BSE, previously buried carcasses may be exhumed. 

Disposal of carcasses and/or offal from sampled animals that have inconclusive or positive test results 

should also follow guidelines provided in the 2019 BSE Response Plan.
29

Attachment A: BSE Ongoing Surveillance Targeting Criteria Flowchart 
30
31

Attachment B: USDA BSE Surveillance Submission Form (VS Form 17-146)
32
33

Attachment C: USDA BSE Surveillance Submission Continuation Form (VS Form 17-146A) 
34

Attachment D: USDA BSE Surveillance Data Collection Form (VS Form 17-131) 
35

Attachment E: IATA 650 Instructions 

https://www.iata.org/contentassets/b08040a138dc4442a4f066e6fb99fe2a/dgr-61-en-pi650.pdf
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37
38

Attachment F: BSE Sample Collection Requirements
39
40

Attachment G: BSE VSLS Quick Start Guide
41
42
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44
45