21 CFR 558.

15 Studies:
A brief history

Jeffrey M. Gilbert, Ph.D.

Microbial Food Safety Team (HFV-157)

Center for Veterinary Medicine

Situation in the 1960s

Issue: What is the impact of antimicrobial
drug use in animals on the potential
development of antimicrobial resistant,
zoonotic foodborne pathogens, and their
subsequent transmission to humans as food
contaminants?

In the 1970s...
Antibiotic in Animal Feed Task Force (FDA)
The task force was formed to address safety
and effectiveness issues associated with
antibiotics administered in animal feed.

Task force findings:

Therapeutic Abx used to relieve animal
disease thought to pose a small risk (high
dose/short duration in young animals)
The benefits to animals were thought to
outweigh potential risks to humans
Identifiable need for using Abx in animals to
relieve pain and suffering
Healthy animals = safer food supply

Task force conclusions:

Pre-approval studies are needed to support
microbiological safety of Abx in foodproducing animals intended for subtherapeutic uses only, including growth
promotion and feed efficiency.

Title 21 CFR Part 558.15

1) Sponsors of Abx are required to submit
study results demonstrating that their
product does not promote bacterial drug
resistance, only when their product is
intended to be administered for greater than
14 days, and for non-prescription use in
food-producing animals

21 CFR 558.15 (b) (1) - (b) (3)

2) Sponsors are required to submit all
information to the Agency on the impact of
their drugs on the Salmonella reservoir in
food-producing animals by specified dates
depending on the drug class

21 CFR 558.15
by July 19, 1973...
records and reports of completed,
ongoing, or planned studies, including
protocols, on the tetracyclines,
streptomycin, dihydrostreptomycin,
penicillin, and the sulfonamides

21 CFR 558.15
by October 17, 1973...
...records and reports of completed,
ongoing, or planned studies, including
protocols, on all other antibiotics

21 CFR 558.15
by March 4, 1974...
...records and reports of completed,
ongoing, or planned studies, including
protocols, on nitrofurans

21 CFR 558.15
by April 20, 1974...
...data from completed studies on the
tetracyclines, streptomycin,
dihydrostreptomycin, the sulfonamides, and
penicillin assessing the effect of the
subtherapeutic use of these drugs in the feed
on the Salmonella reservoir in the target
animal as compared to that in nonmedicated controls

21 CFR 558.15
by April 20, 1975...
...data satisfying all other specified criteria
for safety and effectiveness, including the
effect on the Salmonella reservoir for any
antibiotic or sulfonamide drug approved for
sub-therpeutic use in animal feeds

21 CFR 558.15
by September 5, 1975...
...data satisfying all other specified criteria
for safety and effectiveness, including the
effect on the Salmonella reservoir for the
nitrofuran drugs approved for subtherapeutic use in animal feeds

21 CFR 558.15
Study design
Studies were a set, including a shedding and
resistance component
Studies included a negative control and a
treated group
Animals were inoculated with a lab strain of
Salmonella typhimurium (NAL resistant)
Strain free of transferable R elements

21 CFR 558.15
Study design (contd)
Salmonella were enumerated and tested for
susceptibility
E. coli were tested for susceptibility
Studies were generally 8 weeks long
Test animals were not required to be near
market age or weight

Parameters examined were:

Drug effect on pathogen quantity,
prevalence, and duration of shedding
Drug effect on Salmonella antibiotic
susceptibility
Drug effect on resident E. coli antibiotic
susceptibility

FDA guidance on 558.15 studies

Guideline 18 - Human Health Safety Criteria
Guideline 19 - Animal Health Safety Criteria
Guidelines were the product of the task force
Guidelines can be found at:
http://www.fda.gov/cvm

Integrity measurements
evaluated:

Did the product have Abx properties?

Cross-contamination?
Animal numbers sufficient?
Enough drug consumed to test highest
proposed dose?
Any other drug(s) given?
Any naturally occurring Salmonella present?

Integrity measurements evaluated

(contd)

Salmonella marker stable?

Could Salmonella receive R factors?
Could Salmonella colonize the animals?
Appropriate micro methodology used?
What tissues were examined?
How often were samples taken?
Was study length adequate?

RESULTS
(TOTALS reflective of final decision)
Drug class
Total
Drug/Animal Pass Fail
Reject
____________________________________________________________
Macro/Linco
9
4/3
4
2
3
Ionophores
13
7/3
10
0
3
Unclassified Gm+
15
6/3
9
2
4
Streptogramins
1
1/1
1
0
0
Glycopeptides
2
2/1
1
0
1
Bambermycins
2
1/2
2
0
0
Broad spectrum
2
2/2
1
1
0
____________________________________________________________
TOTALS
44
28
5
11

Studies REJECTED because...

Salmonella or coliform susceptibility
results not submitted
Susceptibility test QC inadequate
Shedding too long to measure prevalence or
duration
Environmental control animals
contaminated or not included

Studies REJECTED because...

(contd)
Animals failed to meet inclusion criteria
Data too disorganized to interpret
Too few animals in study

Problems identified following

retrospective analysis:
Drug spectrum matched Salmonella and
E. coli in 2/44 study sets
Limited info on susceptibility changes in
naturally occurring flora
Artificially high inocula
Lab strain of Salmonella not representative
Small numbers of animals tested

Summary
Most studies that failed did so due to
pathogen shedding
Problems with design and interpretation
Based on policy and regulation of the time
History helpful in steering current and
future efforts on this topic