Quality control and safety of animal products

M. A. Miller
Human Food Safety and Consultative Services, Office of New Animal Drug Evaluation, Center for Veterinary
Medicine, US Food and Drug Administration, Rockville, MD 20857, USA. Received 29 January 1999, accepted 8
September 1999.

Miller, M. A. 1999. Quality control and safety of animal products. Can. J. Anim. Sci. 79: 533538. This paper discusses the
new animal drug approval process regulated by the Center for Veterinary Medicine (CVM), Food and Drug Administration (FDA)
Can. J. Anim. Sci. Downloaded from www.nrcresearchpress.com by 117.239.132.58 on 06/08/17

of the United States. The Center for Veterinary Medicine of FDA considers two criteria in ensuring the human food safety of edible
animal products: i) safety of the chemical residues and ii) for antimicrobial products, microbiological safety including changes in
bacterial pathogen load and resistance pattern. The hazard associated with animal drug products of non-carcinogenic compounds
is assessed by conducting a standard battery of toxicology test, whereas the hazard from the carcinogenic potential of compounds
is evaluated based on structure, results of genetic toxicity tests, and toxicology studies. Post approval monitoring is carried out to
ensure that the animal drugs are being used properly after their approval. Particular concern is given to those eliciting an acute
toxic reaction at relatively low levels. The other aspect of food safety regulated by CVM of FDA is microbiological safety, espe-
cially to antimicrobial drugs used at subtherapeutic levels in feeds. The studies are designed by FDA to ensure that antibiotic treat-
ment of food-producing animals does not alter pathogen load or resistance pattern of pathogens. Two studies are generally
performed: i) the salmonella shedding study, which addresses the effect of drug treatment on the excretion of salmonella in the
feces of animals artificially infected with salmonella; and ii) the coliform resistance study, which monitors the effect of the drug
on the resistance pattern of E. coli present in the endogenous fecal flora. After a retrospective study of the microbiological safety
over past 20 yr, CVM of FDA is planning to revise some microbiological safety studies with focuses on: i) pathogen load, pathogen
excretion and microorganism resistance pattern at the time of slaughter; and ii) susceptibility studies on products that have utility
in human medicine.
For personal use only.

Key words: Animal drug, food safety, antibiotic

Miller, M. A. 1999. Contrle de qualit et scurit des produits animaux. Can. J. Anim. Sci. 79: 533538. Dans cette commu-
nication nous scrutons le nouveau protocole dapprobation des mdicaments pour animaux rgi par le Centre for Veterinary
Medicine (CVM), lequel relve de la Food and Drug Administration (FDA) des tats-Unis. Dans sa tche de veiller la scurit
pour les consommateurs des produits animaux comestibles, le CVM prend en considration deux critres 1) linnocuit des rsidus
chimiques et 2) en ce qui concerne les produits antimicrobiens, la scurit microbiologique, y compris les modifications de la
charge de bactries pathognes et les phnomnes de rsistance. Les risques rattachs aux substances mdicamenteuses non can-
crignes sont valus au moyen dune batterie de tests toxicologiques standards, tandis que lvaluation du pouvoir cancrogne
dun produit repose sur des tudes structurales ainsi que sur des preuves de toxicit gntique et sur des tests toxicologiques. La
surveillance post-approbation voit ce que les mdicaments pour animaux soient utiliss bon escient. Une attention particulire
est accorde aux mdicaments qui provoquent une raction toxique aigu des doses demploi relativement basses. Lautre sujet
de surveillance du CVM est la scurit microbiologique, en particulier celle lie lemploi des antimicrobiens des doses sub-
thrapeutiques dans les aliments des animaux. On veille ce que le traitement aux antibiotiques des animaux destins lalimen-
tation ne modifie pas les populations de pathognes ou leur comportement de rsistance. Deux tests sont couramment utiliss i) le
test dexcrtion de salmonelles, par lequel on observe leffet du traitement mdicamenteux sur le niveau dexcrtion de salmo-
nelles dans les fces des animaux artificiellement salmonelliss. Deuximement le test de la rsistance des coliformes, qui surveille
lvolution de comportement de rsistance de E. coli, prsent dans la flore fcale endogne sous leffet dun mdicament. Aprs
une rtrospective sur ltat de la scurit microbiologique au cours des 20 dernires annes, le CVM se propose de rexaminer les
mthodes de surveillance microbiologique, notamment en ce qui a trait i) la charge en organisme pathogne, lexcrtion des
pathognes et le comportement de rsistance microbiologique au stade de labattage des animaux viande et ii) aux tudes de sen-
sibilit sur les produits qui revtent une utilit en mdecine humaine.

Mots cls: Mdicaments pour les animaux, scurit des aliments, antibiotique

The United States Food and Drug Administration is the
primary federal agency responsible for the regulation of
food and drug products intended for use in humans and
animals. While the Center for Food Safety and Applied
Nutrition regulates the vast majority of human food, the
Center for Veterinary Medicine ensures the human food
safety of animals drug products that enter the human food
supply as residues in milk, meat and eggs produced from
treated animals. While CVM is responsible for performing
the safety assessments for animal drug residues, the United
533
States Department of Agriculture, Food Safety Inspection
Service is primarily responsible for testing the meat supply
for microbiological contamination and animal drug residues.
In fact, both FDA and USDA have extensive programs to
Abbreviations: ADI, acceptable daily intake; CVM,
Center for Veterinary Medicine; FDA, Food and Drug
Administration; INAD, Investigational New Animal Drug;
MRL, maximum residue limits; NADA, New Animal Drug
Application; NOEL, no observed effect level
 534 CANADIAN JOURNAL OF ANIMAL SCIENCE
ensure that meat, milk and other animal products are safe,
wholesome and of high quality.
New Animal Drug Approval Process
Before any animal drug is legally marketed for use in food-
producing animals, the drugs sponsor must have a New
Animal Drug Application (NADA) approved by the FDA.
To obtain a NADA approval, the drug sponsor must demon-
strate to the FDA that the drug is effective and safe in the
target animals, safe for humans consuming edible products
from treated animals, safe for the environment and can be
manufactured to uniform standards of purity, strength and
Can. J. Anim. Sci. Downloaded from www.nrcresearchpress.com by 117.239.132.58 on 06/08/17
identity. Under the federal Food Drug and Cosmetic Act
(the Act), animal drugs mean articles intended for use in the
diagnosis, cure, mitigation, treatment or prevention of dis-
ease in animals and articles (other than food) intended to
affect the structure or any function of the body of animals
(Anonymous a). To conduct investigational studies with
new animal drugs before approval, the drug sponsor must
request an Investigational New Animal Drug (INAD)
exemption from the FDA. Under the INAD exemption, the
drug sponsor can conduct safety and efficacy studies
required for approval of their product. Relatively early in the
investigational process for a drug used in food animals,
CVM scientists review studies relating to human food safety
and establish an appropriate period for drug withdrawal
For personal use only.
before slaughter to ensure that no unsafe residues are pre-
sent in the food products. Once the human food safety is
confirmed, the FDA may authorize the use in human food of
products from animals treated in investigational studies
(Anonymous 1997a).
Human Food Safety Requirements
Whenever an animal is treated with an animal drug, some
amount of the drug (i.e., residue) remains in edible tissue of
the treated animal. Drug residue is defined as any com-
pound present in edible tissue of the treated animals that
results from the use of the drug including the drug itself, its
metabolites and any substances formed in or on the food as
a result of drug use. Substance can be broadly defined to
include everything from drug metabolites to resistant bacte-
ria (Anonymous a). However, generally, CVM considers
two criteria in ensuring the human food safety of edible ani-
mal products: 1) safety of the chemical residues including
the parent drug and all metabolites; and 2) for antimicrobial
products, microbiological safety including changes in bacte-
rial pathogen load and resistance pattern that occur as a
result of drug use in food producing animals.
Although food safety has always been a major concern of
the FDA, the human food safety requirements for animal
drug residues have changed over the years. Originally, ani-
mals drugs were approved based on a No residue or ones lifetime. The amount of drug residue permitted in each
Zero tolerance policy. While the No residue concept of the edible tissues, i.e., the safe concentration, depends
was generally acceptable to consumers and politicians, the
zero tolerance actually represented the sensitivity of the
analytical method used to monitor drug residues not a true
zero value. In fact, because most drugs are eliminated from
the body by first order kinetics, pharmacokinetic theory pre-
dicts that some drug residue would always remain in animal
Table 1. Standard battery of toxicology tests
I. 90-day feeding study in rodent and non-rodent species
II. Multigeneration reproduction study with teratology component
III. Battery of genetic toxicology tests
Table 2. Consumption values for edible tissues
Muscle 300 g
Liver 100 g
Kidney, Fat 50 g
Milk 1.5 L
Eggs 100 g
tissues. As analytical methods improved, the zero toler-
ance was continually needing to be lowered.
To correct this problem, the agency adopted a negligi-
ble tolerance policy. Under this policy, the amount of tox-
icology data required for approval was limited to subchronic
studies in dogs and rats; and the tolerance was generally
assigned as 0.1 ppm in all edible tissues. As with the zero
tolerance procedure, the negligible tolerance was not
related to the potential hazard of the compound. Thus, there
was a disconnect between the permitted residue level or tol-
erance and public health concerns for the drug.
To remedy this situation, the Center adopted its current
risk assessment procedure whereby the tolerance is based on
toxicology, residue and metabolism data. The current risk
assessment model is: the risks from animal drug residue
equals the hazard of the drug product times the exposure to
the drug residues. The agency regulates the public health
risks associated with animal drug residues by assessing hazard
and controlling exposure to meet a risk standard of reason-
able certainty of no harm.
Hazard Assessment: Standard Battery of
Toxicological Tests
The hazard associated with an animal drug product is
assessed by having the sponsor conduct a standard battery of
toxicology tests (Table 1). Each one of these toxicology
tests is designed to examine a different endpoint and deter-
mine a dose at which the drug causes an effect and a dose at
which the drug produces no observed effect. In determining
the toxicological endpoints to be examined, the battery of
tests focuses on the adverse effect of chronic, low-level
exposure to drug residue (FDA 1994).
For compounds that are not carcinogenic, the no observed
effect level (NOEL) of the most sensitive effect from the
most appropriate toxicology studies is divided by a safety
factor to determine an acceptable daily intake (ADI). The
ADI represents the total drug residues, parent and all
metabolites, that can be safety consumed daily throughout
upon the quantity of tissue that is consumed by the 90th per-
centile consumer on a daily basis (FDA 1994) (Table 2).
Controlling Exposure I: Residue and Metabolism
Once the safe concentration is determined, the risk to con-
sumers is minimized by controlling exposure. Generally, the
 MILLER QUALITY CONTROL AND SAFETY OF ANIMAL PRODUCTS
tissue concentrations of drug are highest immediately after
treatment. Also, edible organ tissues, i.e., liver and kidney,
contain more drug residues than muscle. With time, the ani-
mals normal metabolic and excretory processes remove the
drug from the animal. Thus, the first step in controlling
exposure is to determine when the level of drug in the ani-
mals reaches the calculated safe concentration for each of
the different edible tissues.
The first residue chemistry study conducted is the total
residue and metabolism study. This study utilizes radio-
labeled drug and is therefore capable of monitoring all drug-
derived residues resulting from the administration of the test
Can. J. Anim. Sci. Downloaded from www.nrcresearchpress.com by 117.239.132.58 on 06/08/17
compound. In this study, the radiolabeled drug is administered
to a group of animals for which the product is intended.
Animals are killed and the tissues analyzed at various times
after drug administration to determine where the drug
deposits, how it is metabolized and how it depletes from the
animal. Usually, the total radioactivity in the edible tissues
is measured by combustion of the tissue samples to liberate
the radiolabel followed by liquid scintillation counting.
Since the only source of radioactivity is the administered
drug, any detected radioactivity is drug-related (FDA 1994).
Metabolism of the drug is evaluated by extracting the
radioactive material from the tissue followed by chromato-
graphic analysis. By measuring the concentration of the
major radiolabeled metabolites isolated from the tissues col-
For personal use only.
lected at difference times following treatment, it is possible
to determine the drug residue that persists the longest and
the edible tissue that eliminates the drug most slowly.
Because the radiolabeled residues represent all the drug
present in edible tissue, these values can be compared with
the safe concentration directly. When the total residue in
edible tissue is less than half the respective safe concentra-
tion, no preslaughter withdrawal period is assigned. In all
other cases, the drug depletion profile is used to determine
the withdrawal period, i.e., the amount of time needed to
allow the total residue to deplete below the safe concentra-
tion in the tissue that depletes the slowest.
The total residue consists of parent compound, free
metabolites, and metabolites that are covalently bound to
endogenous molecules, and is very difficult to measure on a
routine basis. Rather than measuring the total residues in all
the edible tissues, FDA establishes one value, a tolerance, in
one tissue, the target tissue, for monitoring drug residues.
The tolerance is based upon the marker residue rather than
the total residue. The marker residue may be parent, any
metabolite or a combination of residues that is in known
relationship to the total residue and is measured by the reg-
ulatory assay. The tolerance is established so that when the
marker residue is below the tolerance in the target tissue the
whole carcass is safe. If an animal drug product requires
time after the last treatment to deplete to the calculated safe
concentration, the FDA establishes a withdrawal time for
the product. To establish a withdrawal time, the drug spon-
sor conducts a residue depletion study using the commercial
product in field use conditions. The withdrawal time is cal-
culated by determining the 99th percentile tolerance limit
with 95% confidence. Using this procedure there is only a
5% probability that 1 animal in 100 could have residues
535
above the tolerance at the end of the withdrawal period.
Most animals are slaughtered at market weight not at the
end of the withdrawal period, virtually no violative residues
will occur when drugs are used according to the label.
Information about the withdrawal time is included on the
drug label (FDA 1994).
Regulation of Carcinogens
The Delaney Clause of the FD&C Act prohibits the use of
compounds found to induce cancer when ingested by man
or animal, or if it is found, after tests which are appropriate
for the evaluation of the safety of the food additive, to
induce cancer in man or animal (Anonymous b). An excep-
tion to the Delaney Clause, known as the DES Proviso,
exists for animal drugs, when it is determined by methods of
examination prescribed or approved by the Secretary (...)
that no residue of that compound will be found in the food
produced from those animals under conditions of use rea-
sonably certain to be followed in practice (Anonymous
1997b).
To implement the DES Proviso of the Delaney Clause,
FDA relies upon quantitative risk assessment procedures to
determine when the amount of residue of the carcinogenic
concern present in edible tissue produces an incremental
estimated cancer risk for a lifetime of 1 in 1 million. This
level is functionally defined as no residue (FDA 1987).
The carcinogenic potential of a compound is evaluated
based on structure, results of genetic toxicity tests, and toxi-
cology studies. If the compound or its metabolites are struc-
turally related to a demonstrated carcinogen, it is assigned to
a category with the highest concern for carcinogenic risk. If
further testing in short-term genetic toxicity tests and sub-
chronic feeding studies do not indicate carcinogenicity, use
and exposure levels of the compound are considered to
determine if chronic rodent carcinogenicity studies are
required (Guyer et al. 1994).
When the results of chronic bioassays demonstrate that a
compound induces cancer by the oral route, a nonthreshold,
conservative, linear-at-low-dose extrapolation procedure is
used to estimate an upper limit of low-dose-risk (Lorentzen
et al. 1997). This no residue value is adjusted for con-
sumption and is used to determine how sensitive the analyt-
ical procedure must be to ensure the safety of the food.
Following a determination of no residue, residue and
metabolism studies are conducted as described above.
International Food Safety Standards
Most industrialized countries have regulatory bodies, like
the FDA, that perform similar risk assessments and set tol-
erances or maximum residue limits (MRL) for animal drug
residues in edible tissues. While risk assessment procedures
are similar, countries differ in the toxicology tests required,
the safety factors applied to toxicology tests, their assump-
tions on food commodities consumed per day, and the
amount of the ADI allocated into the MRL.
Codex Alimentarius Commission (CODEX)
In 1962, the CODEX was established as a subsidiary of the
Food and Agricultural Organization and the World Health
 536 CANADIAN JOURNAL OF ANIMAL SCIENCE
Organization of the United Nations. CODEX was created to
facilitate international trade by developing international
food standards. The Codex Commission on Residues of
Veterinary Drugs in Foods provides expert advise on tech-
nical barriers to trade for veterinary products and is respon-
sible for establishing international MRLs for veterinary
drugs. CODEX MRLs are not mandatory but are established
as the standard for trade negotiations. In establishing the
MRLs, the Codex Commission on Residues of Veterinary
Drugs in Foods receives advice from the FAO/WHO Joint
Expert Committee on Food Additives. This committee of
international experts on toxicology, pharmacology and
Can. J. Anim. Sci. Downloaded from www.nrcresearchpress.com by 117.239.132.58 on 06/08/17
metabolism reviews data similar to that reviewed by the
FDA and establishes international ADIs and MRLs for drug
residues. Countries that do not have regulatory bodies over-
seeing animal drug use can adopt the CODEX standard and
be assured that their animal-derived food products are safe
and permitted in international trade.
Controlling Exposure II: Post Approval Monitoring
In addition to establishing a tolerance or MRL for drug
residues, countries must have a monitoring program to
ensure that the animal drugs are being used properly. There
is no international program for assuring that animal drugs
are being used properly; this function is performed by
national regulatory authorities. In the US, the proper use of
For personal use only.
the animal drug product is ensured by providing appropriate
use information on the label, educating animal producers
and veterinarians about proper drug use and monitoring the
food supply for violative drug residues. The Food Safety
Inspection Service of USDA oversees the residue monitor-
ing program in the US and their monitoring data from random
samples indicate that violative drug residues occur very
infrequently, i.e. less than 1% of the nations food supply.
FDA is responsible for taking regulatory action when a
violative residue is identified. FDA prioritizes its regulatory
action based on public health concerns. While the health risk
assessment for animal drug residues focuses on the chronic
low-level exposure effects of drug residues, regulatory com-
pliance actions on violative samples focus on the acute
effects of drugs. Thus, FDA is particularly concerned about
drug residues that elicit an acute toxic reaction at relatively
low doses.
Acute Toxicities Associated with Animal Drugs
Allergic reactions are manifested in many ways, from life-
threatening anaphylactic reactions to lesser reactions such as
rashes. Although animal drug residues probably do not
cause primary sensitization of individuals because expo-
sures are too low and for short duration, violative levels of
animal drug residues in food have caused allergic reactions
in sensitized individuals (Paige et al. 1997).
Violative levels of penicillin are the most frequently cited
cause of allergic reactions in persons consuming residues
probably because a large number of individuals are allergic
to penicillin, allergic reactions occur at low doses and peni-
cillin is extensively used in food producing animals. Many
other animal drugs, including tetracyclines, sulfonamides
and aminoglycosides, can cause allergic reactions in sensi-
tized individuals (Paige et al. 1997).
Violative levels of certain animal drugs can induce phar-
macological effects in consumers. For example, the beta-
agonist, clenbuterol, was approved for therapeutic use as a
bronchial dilator in some food-producing animals in
Europe. There have been several reports of people becom-
ing ill after consuming beef products containing clenbuterol
residues. Clinical symptoms include muscle tremors, heart
palpitations, nervousness, chills and vomiting, i.e., symp-
toms characteristic of the pharmacological activity of beta-
agonists (Paige et al. 1997).
Also, violative levels of antimicrobial animal drugs may
perturb the human gut microflora. The indigenous bacteria
that populate the GI tract of humans provide resistance to
infections, metabolize toxins and carcinogens, produce vita-
mins and aid in food digestion. Therapeutic levels of some
antibiotics perturb the gut ecosystem causing adverse health
effects. To date, there have been few studies to determine
the lowest dose of antibiotic required perturbing the normal
human flora. While it is generally agreed that the very low
tolerance levels of antibiotic residues in food are not suffi-
cient to affect the flora, violative levels of antibiotics may
perturb this ecosystem (Paige et al. 1997).
FDA takes regulatory action, including criminal prosecution,
against individuals that misuse animal drug products when
these acute toxic effects represent a public health concern.
Microbiological Safety
The other aspect of food safety regulated by CVM is micro-
biological safety. Although the microbial foodborne disease
burden of the US is not known with accuracy, it is estimated
that from 6.5 to 33 million cases occur annually and that
deaths from foodborne disease may be as high as 9000 per
year. Foods of animal origin are often identified as the vehi-
cles of food borne disease; and therefore some portion of
these foodborne diseases may be attributed to pathogens
present in the animals colon at the time of slaughter when
feces containing pathogens may contaminate the carcass.
Approximately 1% of the beef carcasses are contaminated
with Salmonella, while contamination of chicken carcasses
is 20-fold higher (US Department of Agriculture 1996).
FDA/CVM began requiring specific antimicrobial studies
in the mid-1970s based on the concerns identified by FDAs
Task Force on Antibiotics and Animal Feed. This Task
Force identified six safety criteria which must be met by
antimicrobial drugs used at subtherapeutic levels in feeds
prior to their approval. According to these criteria, the
antimicrobial drug shall not result in 1) a significant
increase in the frequency of antimicrobial resistant coliform
bacteria in the gut, 2) increased shedding of resistant
Salmonella in animals, 3) increased virulence/pathogenicity
of bacteria, 4) animal disease that is more difficult to treat,
5) residues in edible tissues that are high enough to select for
resistant bacteria when consumed by humans and 6)
increased frequency of hypersensitivity reactions in
humans. The last two criteria were applied to both therapeu-
tic and subtherapeutic products because these concerns are
the same for both classes of products (FDA 1977a, b).
The first four criteria were applied to subtherapeutic
products because data indicated these products presented a
 MILLER QUALITY CONTROL AND SAFETY OF ANIMAL PRODUCTS
greater risk. Subtherapeutic uses involved administering
low levels of antimicrobials to large numbers of animals for
prolonged periods of time in some cases for a lifetime.
Several studies showed that duration of treatment was as
important as dose in the selection of resistant organisms.
Therefore, preapproval testing requirements to support
antimicrobial products were only required for those antibi-
otics which were administered in animal feed for subthera-
peutic purposes including growth promotion and feed
efficiency claims. Subtherapeutic treatment was defined as
treatment for more than 14 d. It was assumed that any
responses seen during the study would be maintained until
Can. J. Anim. Sci. Downloaded from www.nrcresearchpress.com by 117.239.132.58 on 06/08/17
slaughter and negatively impact consumers if edible tissues
become contaminated with pathogens and/or resistant bacteria
(Cooper 1997).
Testing Design for Microbiological Studies
The studies designed by FDA scientists to ensure that anti-
biotic treatment of food-producing animals does not alter
pathogen load or resistance pattern of pathogens have been
summarized in detail (FDA 1977a,b). Two studies were
generally performed; one study addressed the effect of drug
treatment on the excretion of Salmonella in the feces of ani-
mals artificially infected with Salmonella , i.e., Salmonella
shedding studies. The other study, a coliform resistance
study, monitored the effect of the drug on the resistance pat-
For personal use only.
tern of E. coli present in the endogenous fecal flora.
The studies were designed to last 8 wk with animals
receiving drug for the length of the study. Study duration
was based on earlier studies, which suggested that at least
22 d were needed to detect changes in susceptibility pat-
terns. In each study, the drug-treated animals had to ingest
or receive by bolus at least 75% of the highest intended
antimicrobial treatment dose each day of the study including
days when the animals would not eat. The number of ani-
mals in each group ranged from 7 to 12 across all species.
Animal numbers were considered acceptable if the differ-
ence among the animals within a group did not exceed the
difference among the animals between the groups. The ani-
mals were usually housed individually to avoid biasing the
study through bacterial cross-contamination and to increase
the statistical power of the study (FDA 1977a). While these
studies were originally intended to apply to all drugs intend-
ed for subtherapeutic use in any animal species; cattle,
swine, and poultry were the only species tested (Cooper
1997).
In the shedding study, treated and control animals were
inoculated a couple of days before the start of drug treat-
ment with Salmonella at a sufficient concentration and
under conditions to induce initial shedding at about 105
CFU mL1 (i.e., up to two inoculations at approximately 109
CFU mL1 concurrent with starvation). The purpose of this
artificially high inoculation dose was to create an initial
infection sufficient to detect a 2 log difference in Salmonella
excretion due to the drug. In cases where shedding was too
heavy to determine duration at the end of 8 wk; a second
shedding study was performed at a lower inoculum density,
i.e., approximately 108 CFU mL1. All shedding studies
included an environmental control group, which was not
537
inoculated or given test drug. Since the test drug was deter-
mined to have no effect when the results from the drug-
treated and control group were the same, the environmental
control was used as a sentinel to detect if bacterial cross-
contamination between the study groups contributed to the
lack of difference (FDA 1977a).
The drug-treated and control groups in the Salmonella
shedding studies were inoculated with a laboratory strain of
S. typhimurium. The strain used was known to colonize the
test species, was easily recovered because it carried chro-
mosomal nalidixic acid resistance, was susceptible to 8 to 10
medically-relevant antimicrobial drugs, and was a capable
recipient of resistance-carrying plasmids. Because it is diffi-
cult to detect changes in bacterial susceptibility following
drug treatment, it was preferred that animals in the shedding
study carry high levels of resistance in their coliform popu-
lation. It was presumed that with more donor bacteria pre-
sent in the coliform population, it would be more likely that
Salmonella would receive resistance factors. The intent of
using a Salmonella strain free of plasmid resistance was to
determine the rate of transfer of resistance factors due to
drug effects (FDA 1977a).
Feces were collected at approximately weekly intervals.
Tissues, e.g. mesenteric lymph node and spleen, and cecal
contents were collected at the end of the study. Shedding
quantity, duration, and prevalence in animals as well as tis-
sue invasion and Salmonella susceptibility results from the
drug- treated and control groups were compared.
Susceptibility test quality control results were evaluated to
determine if methodology was appropriate (FDA 1977a).
The design for the coliform resistance study was essen-
tially the same as that described for the Salmonella shedding
studies except that animals were not artificially inoculated
with bacteria. Rather, the effect of drug on the endogenous
fecal flora was evaluated. Because it is difficult to detect a
change in resistant E. coli if the majority of the population
is resistant before drug exposure, for the coliform resistance
study, it was necessary to use animals with a low level of
resistance (less than 20%) in their endogenous E. coli.
Changes in coliform susceptibility (against the same med-
ically relevant drugs used in the Salmonella shedding studies)
between the drug-treated and control groups indicated drug
effects on either resistance transfer or new emergence. Thus,
the results of this study were coupled with the results of the
Salmonella shedding study to determine if the drug effects
were on the selection of resistant genes or on the transfer of
resistance factors (FDA 1977a,b).
Retrospective Analysis of the Microbiological
Safety Studies
The Center for Veterinary Medicine has recently completed
a retrospective analysis of all the data that have been sub-
mitted over the past 20 yr and is planning to revise these
microbiological safety studies to better suit todays needs.
Because of changes in antimicrobial product usage and
spectrum of activity, the current studies have some weak-
nesses. For example, the microbiological safety studies, as
designed, focus more on environmental or occupational
safety concerns, i.e., the bacteria that are shed on the farm,
 538 CANADIAN JOURNAL OF ANIMAL SCIENCE
than on food safety aspects, i.e., the bacteria present at the
time of slaughter. Food is distributed throughout the coun-
try and foodborne pathogens are more likely to affect large
portions of the population compared with environmental or
occupational routes of exposure.
Studies will be redesigned to focus on pathogen load,
pathogen excretion and microorganism resistance pattern at
the time of slaughter, since it is organisms present in the ani-
mal at the time of slaughter that have the greatest potential
to contaminate the food supply.
The Center for Veterinary Medicine will focus the sus-
ceptibility studies on products that have utility in human
Can. J. Anim. Sci. Downloaded from www.nrcresearchpress.com by 117.239.132.58 on 06/08/17
medicine. Drugs tested under the old criteria included
ionophores, bambermycins, and other unclassified gram-
positive drugs that have no utility in human medicine. None
of these drugs changed the susceptibility pattern of coliforms
or Salmonella in the traditional studies. Also, there was no
demonstration in the literature of transferable resistance to
drugs in these pharmacological classes. In the future, sus-
ceptibility studies may not be needed for drugs similar to
these products and with no utility in human medicine.
In the traditional microbiological studies, Salmonella and
E. coli were the only pathogens investigated for changes in
susceptibility. These pathogens are gram-negative bacteria,
and many antimicrobials given to animals have activity
against gram-positive organisms only. In the future, pathogen
For personal use only.
testing will be matched to the spectrum of drug activity.
That is, Salmonella and E. coli will continue to be evaluated
for drugs possessing activity against a broad spectrum of
organisms and gram-negative organisms; but enterococci
will be evaluated for products with a gram-positive spec-
trum of activity.
The effect of the antibiotic on pathogen load and resis-
tance pattern will be evaluated under proposed conditions of
use for the product. Salmonella inoculation concentrations
will be comparable to levels seen in natural infections, drug
treatments will be similar to those proposed for approval,
and, when appropriate, the effects of a drug withdrawal period
will be assessed. Furthermore, since the retrospective analy-
sis never showed that drug treatment affected tissue colo-
nization by Salmonella, tissue colonization tests will be
required only when there is evidence to suggest a particular
antimicrobial might affect this parameter.
Finally, changes in the use pattern of therapeutic antimi-
crobials may necessitate performing microbiological food
safety studies for some of these products as well as the sub-
therapeutic products. While traditional studies were only
required for antimicrobials administered in feed, studies
may be necessary for other routes of administration with
long-term or widespread exposure.
CONCLUSION
The Food and Drug Administrations Center for Veterinary
Medicine approves the use of safe and effective animal drug
products for food producing animals. Before these products
are approved, CVM ensures the food products from these
animals will be safe and wholesome. After product
approval, FDA/CVM interacts with other federal and state
agency to ensure that the consumer enjoys quality animal
food products.
Anonymous a. Federal Food Drug and Cosmetic Act, Chapter II,
Section 201(g)(1).
Anonymous b. Federal Food Drug and Cosmetic Act, Chapter IV,
Section 409(c)(A)(2).
Anonymous. 1997a. United States Code of Federal Regulations 21
Part 514, revised 1 April 1997.
Anonymous. 1997b. United Stated Code of Federal regulations 21
(Part 500) revised 1 April 1997.
Cooper, J. N. 1997. Perspective on the therapeutic and growth
enhancing use of antibiotics by the livestock industry and the
antibiotic resistance monitoring program. J. Anim. Sci. (submitted).
Food and Drug Administration 1977a. Penicillin in animal feed.
Federal Register 42: 4376943793.
Food and Drug Administration 1977b. Chlortetracycline and
oxytetracycline in animal feed. Federal Register 42: 5625356289.
Food and Drug Administration 1987. Sponsored compounds in
food-producing animals: Criteria and procedures for evaluating the
safety of carcinogenic residues: Animal drug safety policy. Federal
Register 4957249590.
Food and Drug Administration 1994. Guideline for establishing
a safe concentration; Availability. Federal Register 52: FR
3749937500.
Gaylor, D. W., Axelrad, J. A., Brown, R. P., Cavagnaro, J. A.,
Cyr, W. H., Hulebak, K. L., Guyer, C. G. and Miller, M. A.
1994. Human food safety evaluation of repartitioning agents.
Pages 355417 in H. Hafs and R. G. Zimbelman, eds. Low-fat
meat: Design strategies and human implications. Academic Press,
Inc., New York, NY.
Lorentzen, R. J., Miller, M. A., Mulligan, L. T. and Schwetz,
B. A. 1997. Health risk assessment practices in the U.S. Food and
Drug Administration. Regul. Toxic. Pharmacol. 26: 307321.
Paige, J. C., Tollefson, L. and Miller, M. 1997. Public health and
drug residues in animal tissues. Vet Human Toxicol. 30: 162169.
US Department of Agriculture. 1996. Pathogen reduction; hazard
analysis and critical control point (HACCP) systems; Final Rule.
Federal Register 61: 3880638989.