493
Veterinary use and antibiotic resistance
Michael Teuber
Globally, an estimated 50% of all antimicrobials serve
veterinary purposes. Bacteria that inevitably develop
antibiotic resistance in animals comprise food-borne
pathogens, opportunistic pathogens and commensal
bacteria. The same antibiotic resistance genes and gene
transfer mechanisms can be found in the microfloras of
animals and humans. Direct contact, food and water link
animal and human habitats. The accumulation of resistant
bacteria by the use of antibiotics in agriculture and veterinary
medicine and the spread of such bacteria via agriculture and
direct contamination are documented.
Addresses
Laboratory of Food Microbiology, Eidgenössische Technische
Hochschule Zürich, CH-8092 Zürich, Switzerland;
e-mail: teuber@ilw.agrl.ethz.ch
Current Opinion in Microbiology 2001, 4:493–499
1369-5274/01/$ — see front matter
© 2001 Elsevier Science Ltd. All rights reserved.
Abbreviations
STEC Shiga-toxin-producing Escherichia coli
VRE vancomycin-resistant enterococci
Introduction
The development of antibiotic resistance in bacteria has
been attributed to the use of antimicrobials in human
medicine. The contributions of veterinary medicine and
agriculture to antibiotic resistance are still being discussed.
The veterinary use of antibiotics includes the use on pets,
farm animals and animals raised in aquaculture. For farm
animals, antibiotics are used in therapy and prophylaxis,
and to increase growth and feed efficiencies. The main
infectious diseases treated are enteric and pulmonary
infections, skin and organ abscesses and mastitis.
Antibiotics used in both veterinary and human medicine
are: penicillins, cephalosporins, tetracyclines, chloram-
phenicols, aminoglycosides, spectinomycin, lincosamide,
macrolides, nitrofuranes, nitroimidazoles, sulfonamides,
trimethoprim, polymyxins and quinolones [1•]. Of the
more than 1 million tons of antibiotics released into the
biosphere during the last 50 years [2•], roughly 50% are
estimated to flow into the veterinary and agricultural
channels [3]. The application of antibiotics at subthera-
peutical levels for increased growth and feed efficiencies
in farm animals (pigs, cattle, turkeys and chicken), an
integrated part of modern agriculture worldwide, is highly
controversial [4•]. The following antibiotics are approved
for those two purposes in the US: ampicillin, arsenilic acid,
bacitracin, bambermycin, chlortetracycline, dihydrostrep-
tomycin, efrotomycin, lacalocid, monensin, oleandomycin,
penicillin, roxarsone, spectinomycin, tylosin and virgini-
amycin (for details, see [1•,4•]).
The leading textbook on antimicrobial therapy in veteri-
nary medicine proposes that the highest antibiotic
resistance selection pressure occurs in calf, swine and
poultry processing [1•]. It identifies the prevalence of
antimicrobial drug resistance in veterinary pathogens, of
which there is only fragmentary and unbalanced data, to
be one of the areas in which there is a distinct lack of
knowledge. The textbook also states that increasing
controls of antibiotic use in agriculture may require con-
siderable changes in the management of food animals that
are processed under intensive conditions designed to be
highly productive.
Surveillance studies have shown an increased incidence of
resistance development in pathogenic and commensal
bacteria from animals ([5]; for short reviews, see [6•–8•]).
Also, phytopathogenic bacteria have been demonstrated to
become resistant against the antibiotics used in orchards,
an agricultural problem of increased significance outside
the veterinary field [9•].
The medical consequences of antibiotics in agriculture
have been proposed to be the end of the road of the eco-
logical network of resistance: agriculture and veterinary
use contribute to selective pressure, resistance reservoirs
and routes of transmission [10]. The death of two patients
from resistant Salmonella typhimurium DT104 acquired
from pork in Denmark is an example of a sad, though
probably rare, incident [11••].
My main concerns as a food microbiologist are the transfer
and consequences of antibiotic-resistant pathogens and
commensal bacteria into the human population. The
potential routes are manifold (Figure 1).
This review will document: first, the accumulation of
resistant bacteria by the use of antibiotics in agriculture
and veterinary medicine; second, the spread of resistant
bacteria from agriculture via, for example, manure into the
water and soil of the surrounding environment; and third,
the direct contamination of consumers and their digestive
systems with food containing resistant bacteria, including
pathogens of agricultural origin.
How many antibiotics are used?
Statistics on antibiotic sales in the European Union and
Switzerland for the year 1997 from the European
Federation of Animal Health (FEDESA) indicated that
5 460 000 kg of antibiotics were used for human health,
3 465 000 kg for animal health and 1 575 000 kg for animal
growth promotion [12••]. Based on body mass, the antibi-
otic doses available were 241 mg/kg for humans and
54 mg/kg per kg for animals. Large differences in countries
with intensive animal production were noted: Austria,
494 Antimicrobials

Figure 1 animal excrement. Their antibiotic resistance profiles are

indicators of the selection pressure they had to endure and,
under circumstances of contamination, once the resistance
(a) 5 040 000 kg (b) 5 460 000 kg
Antibiotics for Antibiotics for profiles of the contaminant human or animal bacteria are
therapy & therapy & known, the source of the bacteria can be elucidated.
prophylaxis prophylaxis
(3 465 000 kg) or Fecal enterococci and coliforms from surface waters in
growth promotion
(1 575 000 kg) Florida were investigated by discriminant analysis of their
resistance profiles towards nine common antibiotics
(e) Food & direct contact (amoxicillin, ampicillin, cephalothin, chlortetracycline,
oxytetracycline, tetracycline, erythromycin, streptomycin
and vancomycin) [13•]. The average rates of correct
(c) Animals Humans
farm animals hospitals (d)
assignment to human or animal origins of several thousand
pets community enterococi and coliforms of known origin were 62.3% and
fishes 63.9%, respectively. The system was validated in the field:
(f) Sewage Water (g) a ditch contaminated with human effluents contained
human-type fecal enterococci and coliforms and, after
repair, mainly cow and animal ones. Samples from nature
(f) Plants (h) Environment
reserves away from agricultural contamination contained
crops soil
flowers water strains attributable to racoons, rabbits and birds.

Harvest remains
Antibiotics
for therapy &
prophylaxis
(unknown amounts)
Model of the distribution and transfer of antibiotics, antibiotic
resistance genes and microbes in the biosphere, produced in 1997 for
the European Union and Switzerland. Contamination of the biosphere
with antibiotics is represented by straight arrows, and the movement of
resistant microbes and resistance genes is represented by bent
arrows. The circle in the center of the figure symbolizes the microbial
world in which resistance genes are transferred between different
microbes. The main channels of antibiotic input into the biosphere are
treatments of (a) animals [12••] and (b) humans [12••]. (c) Farms
[8•,19•,20,21,22•,23••,24•,25•,26,27•,28••,29•,30,33•,39•,40••] and
(d) hospitals [8•,12••,19•,22•,23••,24•,25•,28••,29•,30,31,37••,40••]
are the quantitatively dominant domains of input. Specific food items
(made from raw meat and milk) and (e) direct contact
[8•,27•,34•,35,36•,37••,38] directly link resistant bacteria in animals
and humans. (f) Fertilization of plants [16•,17•,18,32•] with animal
manure and sewage contaminates salads and vegetables with
resistant bacteria. (g,h) Both humans and animals discharge unused
antibiotics and resistant bacteria into the environment, (i.e. soil and
water [13•,15••,16•]). Modified from [41].
The differences in resistance profiles in this ecological
study clearly reflect the differences in selection pressure in
the investigated geographical location, as has been previously
demonstrated for a watershed in Virginia polluted with
fecal matter of animal origin [14•].
A molecular approach to identifying resistance genes in a
farm environment and in farm effluents is of interest.
Because tetracycline resistance determinants — coded by
tetO, tetQ, tetW, tetM, tetBP, tetS and tetT — confer ribosomal
protection, oligonucleotide primers were devised to target
their genes in the rumina of cows, in swine feed and swine
feces, and in swine fecal streptococci. Identical tetO and
tetW genes were found in swine and cows, tetM only in pigs
and tetQ only in the rumen of cows. The swine fecal
streptococci (mainly Streptococcus alactolyticus) all contained
tetO, and 22% of them contained tetM also. With the exception
of those encoded by tetT and tetBP, all determinants were
found in the commercial components of swine feed (corn,
soybean, whey, plasma products, Tylan and the antibiotic
mixture chlortetracycline, sulfonamide and penicillin) [15••].

When applied to lagoons and underlying ground water of

Denmark, Finland, Ireland and Sweden (6, 24, 24, 12 and
24 mg/kg, respectively) were well below the mean doses,
whereas Spain, Greece and the United Kingdom (103,
134 and 148 mg/kg, respectively) exceeded the mean
doses by far, demonstrating imprudent antibiotic use in
these countries [12••].
Environmental spread of antibiotic resistance
from farm animals
Many food-borne pathogens and opportunistic bacteria
like enterococci have or can have habitats in food animals
(e.g. in the skin and intestinal tract). They can enter meat
and milk products during slaughtering and milking, or
contaminate raw vegetables when the soil is fertilized with
two swine production facilities, this method detected all
eight classes of tet resistance genes in DNA extracts of the
lagoons and the ground water as far as 250 m downstream
of the lagoons. The gene tetM was detected in bacteria
isolated from the ground water (Enterococcus hirae,
Staphylococcus cohnii, Lactobacillus reuteri, Bosea thiooxidans,
Microbacterium oxydans, Afipia spp. and Pseudomonas
pseudoalcaligenes) and some typical soil bacteria. The tet
genes therefore occur in the environment as a direct result
of agriculture. Ground water may be a potential source of
antibiotic resistance in the food chain [16•] (see Figure 1).
Different IncQ plasmids carrying, in different combinations,
resistance against streptomycin, sulfonamide, streptothricin,

kanamycin, tetracycline and chloramphenicol were isolated
from swine and cattle manure in biparental exogenous
matings into recipient Pseudomonas putida UWC1. The
host range included Escherichia coli DH5α, Acinetobacter sp.
BD413, Ralstonia eutropha and Agrobacterium tumefaciens.
Determinants encoded by strA and strB were arranged as in
plasmid RSF1010 (and referred to hereafter as strA–strB),
sometimes coupled with the determinant encoded by
sulII. Other determinants were encoded by catIII, aph
(3′)-Id, sat3 and tetY [17•]. The same strA–strB-encoded
determinants occurred in the streptomycin-resistant
plant pathogen Erwinia amylovora. In contrast to strA–strB
located on non-conjugative small plasmids in bacteria
isolated from humans and animals (e.g. S. typhimurium),
the isolates from the plants E. amylovora, Pseudomonas
syringae and Xanthomonas campestris harboured strA–strB on
large conjugative plasmids within transposon Tn5393 [18].
Foodborne pathogens and antibiotic resistance
Of 50 isolates of Shiga-toxin-producing E. coli (STEC)
from cattle, ground beef and humans (comprising 29 E. coli
0157:H7 STEC and 21 non-0157:H7 STEC), 39 strains
exhibited resistance to two or more antimicrobial classes,
with significant resistance to ampicillin, tetracyclin,
kanamycin, streptomycin and sulfamethoxazole in both
0157:H7 and non-0157:H7 STEC, and to ampicillin-
clavulanic acid, cephalotin, chloramphenicol, forfenicol
and trimethoprim-sulfamethoxazol in the non-0157:H7
STEC. 1 kb and 2 kb integron fragments from the
0157:H7 STEC were amplified with suitable primers and
DNA sequencing, and the resistance genes identified to
be aadA and drfXII. Some integrons were transferable via
conjugation to other 0157:H7 STEC and to several Hafnia
alvei strains. Conjugative tetracycline resistance was
detected, but not within the characterized integrons [19•].
Class 1 integrons and large plasmids from multiresistant
S. typhimurium DT104 have been characterized in hundreds
of Salmonella strains in different countries [20,21,22•].
The ACSSuT resistance phenotype, which is resistance to
ampicillin, chloramphenicol, streptomycin-spectinomycin,
sulfonamide and tetracycline, was characterized in
Canadian isolates. In one isolate, genes encoding resis-
tance to ampicillin (psel), chloramphenicol (pasppflo-like),
streptomycin-spectinomycin (aadA2), sulfonamide (sulI)
and tetracycline (tetG) were mapped to a 13 kb region in
which two copies of sulI were located at the 3′ end of either
the pse1 or aadA2 integrons. The two integrons were sepa-
rated by the pasppflo-like and the tetG genes. A kanamycin
resistance determinant (aphA-1) was either found on a
2 megadalton plasmid or on the chromosome [22•].
Rare but gradually increasing observations of ceftriaxone-
resistant Salmonella infections in the United States have
gained attention. In one case, infection in a 12-year-old
boy was linked to a strain of S. typhimurium variant
Copenhagen, acquired from cattle [23••]. This strain was
resistant to a total of 13 antibiotics. The determinants for
Veterinary use and antibiotic resistance Teuber 495
12 of these were encoded on a 160kb conjugative plasmid.
Resistance included the ACSSuT phenotype. The β-lacta-
mase responsible for ceftriaxone resistance was an AmpC
β-lactamase. The prevalence of such phenotypes in
Salmonella strains increased from 0.1% in 1996 to 0.5% in
1998, 77% of the patients being younger than 18 years [24•].
Another concern is the decreased susceptibility to
ciprofloxacin in S. typhimurium DT104 strains.
Oligonucleotide probes for specific mutations in the gyrA
gene detected Asp87→Asn, Asp87→Gly and Ser83→Phe
mutations in human and animal isolates. Sequencing
revealed two more mutations, Asp87→Tyr and Ser83→Tyr.
The clonal spread of S. typhimurium DT104 is superceded
by the independent appearance of ciprofloxacin resistance
[25•]. An attempt to demonstrate and explain antibiotic-
induced hyperinvasiveness in S. typhimurium DT104
failed, although the Salmonella pathogenicity island 1 was
observed in 400 multiresistant isolates. The putative hyper-
virulence of multiresistant S. typhimurium DT104 seems
not to occur at the level of invasion [26].
Examination of the presence of insertion element IS200-
PCR revealed that cows that shed S. typhimurium heavily
contaminate their environment on the way from the farm
to the slaughterhouse to the final meat product: the transport
equipment, waiting cubicle, and other animals and their
meat product became contaminated [27•].
Campylobacter
In 1997 and 1998 in Spain, Campylobacter jejuni and
Campylobacter coli strains isolated from animal-originated
food and from the feces of broilers, pigs and humans were
found to be 99% (animal) and 71% (human) resistant to
ciprofloxacin, with high cross-resistance to nalidixic acid.
C. coli from pigs had high resistance frequencies to ery-
thromycin (80%) and ampicillin (65%), compared to 34%
and 29%, respectively, in humans. The situation was
declared worrying, as fluoroquinolone resistance was
unknown in Campylobacter in Spain before 1988 [28••]. In
contrast, recent risk assessments of the impact of fluoro-
quinolone-resistant C. jejuni in beef cattle in the USA
determined resistance levels to be around 10% [29•].
Staphylococci
A high percentage of poultry and human staphylococcal
isolates was shown to express erythromycin resistance
owing to the presence of ermA, ermB, ermC and msrA or
msrB genes. The ermA gene was exclusively chromoso-
mal, whereas ermC was found on 2.4–4.2 kb plasmids.
The common restriction fragment length polymorphism
(RFLP) pattern of poultry and human clinical isolates
indicates a common lineage of the ermA gene [30].
Conjugation experiments clearly revealed a high fre-
quency of transfer (4.5 × 10–3) from poultry to human
clinical isolates of tetracycline and ermA or ermC markers.
The mechanisms were identified to be transposition and
transformation [31].
496 Antimicrobials
Enterococci
Enterococci, common intestinal bacteria of animals and
humans, may cause serious nosocomial infections, also
owing to a high level of multiple antibiotic resistance.
Highly efficient broad host range conjugative plasmids and
transposons allow gene exchange between many Gram-
positive and Gram-negative bacterial species in the
intestine. In food, they are regarded as indicators of fecal
contamination There are concerns that enterococci from
animals may transfer all their acquired resistances via food
to human habitats and to human pathogens [8•].
Tn916-like conjugal elements previously detected in
cheese and fermented sausages [8•] have been characterized
by PCR and Southern blots in enterococci from swine
effluent [32•]. Of the investigated enterococci, 71% were
resistant to tetracycline and 34% of those transferred the
traits in mating experiments to Bacillus thuringiensis and
were able to mobilize plasmid pC194 between Bacillus
spp. When inoculated into virgin soil, the swine waste
raised the level of tetracycline-resistant fecal enterococci
from undetectable to 4 × 104 per gram. Over six months,
the level dropped to 5 × 103 per gram only.
Avoparcin can induce cross-resistance to vancomycin. The
ban in Europe on the use of avoparcin as a growth promoter
enabled the investigation of the fate of vancomycin-
resistant enterococci (VRE) in poultry and swine. A
Norwegian study of 109 poultry farms using avoparcin and
33 houses never exposed to avoparcin identified 106 and six
VRE-positive farms, respectively. Resistance mediated by
vanA remained high for more than 18 months after avoparcin
withdrawal (in up to 98% of the Norwegian poultry houses).
In swine, the prevalence of VRE was low (in 1% of herds)
because avoparcin was never used in swine in Norway. The
slow regression of VRE in poultry houses exposed to
avoparcin over nine years reflects the old observation that
antibiotic use eliminates sensitive bacteria, resulting in a
lack of susceptible enterococci to recolonize the flock [33•].
Vancomycin-resistant Enterococcus faecium was also found in
25% of 102 isolates from several industrial and artisan Italian
cheeses (Fontina, Montasio, Taleggio, Grana, Robiola, Italico,
Pecorino, Gorgonzola and Mozzarella). The vanA gene was
identified in every case by PCR [34•].
Antibiotic-resistant enterococci (E. faecalis, E. casseliflavus,
E. faecium and E. durans) were present in food items directly
linking animals and human consumers (135 samples in
total, with 102–107 enterococci per gram): raw milk cheeses
(Emmenthal, Appenzell, Gruyère, Tilsit and soft cheeses),
green salads, mungbean sprouts, minced pork sausages and
raw sausages. Resistance to chloramphenicol, erythromycin
and tetracycline was prominent in E. faecalis. E. faecium
showed resistance to ampicillin, ciprofloxacin, erythromycin,
imipenem, nitrofurantoin, penicillin and tetracycline [35].
Based on common pulsed-field gel electrophoresis
(PFGE) patterns of chromosomal DNA, some highly
kanamycin- and gentamicin-resistant E. faecalis strains
from humans and from cheeses (Morbier, Emmental,
Comté, St. Nectaire and Mamirolle) were found to be
closely related [36•].
E. faecalis and E. faecium from humans, food and starter
cultures were screened for the following virulence
determinants: aggregation protein (agg); extracellular
metalloendopeptidase (gelE); cytolysin precursors (cylLv
and cylLs); post-translational modification of cytolysin
(cylM); transport of cytolysin (cylB); activation of cytolsin
(cylA); cell-wall-associated protein involved in immune
evasion (esp); cell-wall adhesion expressed in serum (efaAfs
and efaAfm); and sex pheromones chemotactic for human
leucocytes (cpd, cob, ccf and cad). E. faecium from starter
cultures and food (cheese, dried milk, canned ham,
uncooked sausage and retail pork) were found to contain
efaAfm. Medical isolates were found to contain esp and,
occasionally, gelE. E. faecalis from food contained, in some
cases, all investigated factors, as did many medical isolates
[37••]. Because antibiotic resistance is so common in food
enterococci [8•,33•,34•,35,36•], it would be surprising if
virulence determinants were not found in these strains.
A large conjugative multiresistance plasmid, pRE25, from
E. faecalis was isolated from a sausage. The 50,237 bp
plasmid is transferable by conjugation to other enterococci,
Listeria innocua and Lactococcus lactis (FV Schwarz,
M Teuber, unpublished data). It is the largest sequenced
Enterococcus plasmid and demonstrates convincingly the
composite nature of resistance plasmids that have collected
genetic information from many different species (see Figure 2).
The multiresistant L. lactis K214 isolated from a raw-milk
soft cheese also has a composite 29,871 bp plasmid [8•]
that carries a determinant for a multiple-drug transporter
protein that confers decreased susceptibility to macrolides,
lincosamides, streptogramin and tetracyclines in L. lactis
and E. coli but not in E. faecalis or Staphlylococcus aureus.
The mdt(A) gene encodes a protein with 12 putative trans-
membrane segments that contain typical motifs conserved
among the efflux proteins of the major facilitator super-
family. In addition, it has two C-motifs (which are part of
the antiporter motif [gX3GPXiGGxl] and highly conserved
in the fifth membrane spanning domain of antiporters
but absent in symporters and uniporters) and putative
ATP-binding sites [38].
Antibiotic resistance in aquaculture systems
Oxytetracycline resistance plasmids of 72 Aeromonas
isolates from a hospital effluent and of 91 Aeromonas isolates
from fish hatchery tanks revealed that 11 and six strains,
respectively, transferred oxytetracycline plasmids to E. coli
J53-1, these plasmids being closely related to IncU plasmids
like pASOT and pRAS1 (from Aeromonas salmonicida
found in fish farms in Norway and Scotland), and pIE420
(from E. coli isolated from a German hospital). pRAS1 and
pIE4120 seemed to be identical — both carried the
 Veterinary use and antibiotic resistance Teuber 497

Figure 2

Genomic structure of the 50,237 bp

PstI
conjugative multiresistance plasmid pRE25 of HindIII
Enterococcus faecalis (GenBank acc. No. EcoRI
EcoRI BglII
X92945) isolated from a sausage. A large PstI BglII
30.5 kb fragment (open reading frames PstI EcoRI
EcoRI HindIII
[ORFs] 6–40), containing the transfer region BglII
(ORFs 24–39, colored orange-pink), is XbaI
homologous to pIP501 from Streptococcus HindIII
AvaII 56 57 58 1 2 3
agalactiae. ORFs 25, 26, 29–32, 34, 36, 37 HindIII
55 4
54 10.1 kb oriR
and 39 code for between one and six 5
53 3.9 kb
transmembrane helices. Antibiotic resistance 6
52
genes are colored bright pink: ORF10 is cat, 51 7
50 50.0 kbp 0.8 kb 8
ORF14 is erm, ORF44 is aadK, ORF45 is HindIII 49 9
sat4 and ORF46 is aph(3′)-III. Three HindIII 48 3.4 kb
replication proteins are present: ORFs 1, 6 HindIII
AvaII 47 10
and 11. Insertion sequences are colored light AvaII PstI
blue. ORFs 5, 41 and 51 are identical with 46 11
AvaII
insertion sequence IS1216V. ORFs 1–5, 45 10.0 kbp
HindIII
40.0 kbp HindIII
41–46, 51–53 and 55–58 are 99%–100% pRE25 12
44
identical on the nucleotide level with 7.4 kb
50,237 bp 13
sequences of the Enterococcus faecium 43
14
Genome Project (United States Department HindIII 42 15
HindIII
of Energy Joint Genome Institute, URL: 16 AvaII
XbaI 41 AvaII
http://www.jgi.doe.gov/tempweb/JGI_microbi 17
al/html/enterococcus/enterococcus_homepag 40 18
39 19
e.html). Genes involved in conjugation are 30.0 kbp 20.0 kbp
KpnI 38
coloured orange-pink; other ORFs are colored HindIII 20
37 HindIII
blue; genes involved in better-than-random 21 HindIII
segregation are colored gray; insertion HindIII 36 22
35 30.5 kb 23
elements (transposases) are colored light EcoRI 34
24
blue; the origin of replication is represented HindIII 33
32 25 oriT
by the brown bar; the origin of transfer is HindIII 31 30 27 26
KpnI 29 28 HindIII
represented by the red bar; segments HindIII
between insertion elements are represented AvaII HindIII
by the circular green bars. Reproduced with HindIII HindIII
permission from [42]. KpnI HindIII
HindIII
HindIII
HindIII

tetracycline resistance transposon Tn1721. Related
tetracycline-resistance-encoding plasmids have been
disseminated between different Aeromonas spp. and E. coli,
and between the human and aquaculture elements in
distinct geographical locations [39•]. The human and the
aquaculture microbial compartments of the environment
are thus not separated (see Figure 1). These observations
prompted the successful shift from antibiotic use to vacci-
nation in Atlantic salmon hatcheries in Norway as a
method of countering the most prevalent bacterial salmon
pathogens, Vibrio salmonicida, Vibrio aguillarum and
A. salmonicida [40••].
water and direct contact can spread these bacteria from
animal microfloras to human micofloras. Elimination of
resistance determinants from these microfloras is slow,
particularly if there is no reservoir of susceptible bacteria
available to recolonize the host animal. One question to
be answered is whether or not commensal bacteria from
food can transfer their resistance genes to the indigenous
human microflora during intestinal transit. Prudent use of
antibiotics is recommended and should be pursued until
we have exact statistics on the quantitative contribution
of veterinary medicine and agriculture to the antibiotic
resistance nightmare.

Conclusions
The molecular analysis of antibiotic resistance genes,
plasmids and transposons has demonstrated that identical
elements are found in animals and humans. The phenom-
enon applies to pathogenic (e.g. foodborne), opportunistic
and commensal bacteria: the use of antibiotics in veteri-
nary medicine, similar to its use in agriculture and
aquaculture, selects for resistant bacteria. These bacteria
are released into the environment, where they can be
easily demonstrated, in animal feces. Specific food items,
Update
VRE carrying the vanA gene [33•] were still prevalent in
Norwegian poultry carcasses even three years after
avoparcin was banned [43•]. In contrast, Aarestrup et al.
[44••] demonstrated the prevalence of glycopeptide-resis-
tant fecal enterococci (E. faecium and E. faecalis) from food
animals (broilers and pigs) in Denmark dropped from
72.7% in 1995, the year avoparcin was banned in Denmark,
to 5.8% in 2000. Likewise, the prevalence of resistance to
the antibiotics avilamycin, erythromycin and virginiamycin
498 Antimicrobials
dropped after government or self-imposed bans on the use
of these antibiotics and tylosin as growth promoters [44••].
This proves that it is possible to reduce the occurrence of
antimicrobial resistance in the microflora of national popu-
lations of food animals when selective pressure is removed.
The same Danish group (M Blom, TL Sorensen,
RL Poulsen, N Frimodt-Moller, DL Monnet, FM Aarestrup,
F Espersen, unpublished data) also presented preliminary
results of an ingestion study in which human volunteers
consumed vancomycin-resistant E. faecium originally isolated
from broiler meat. This random, double-blind study
revealed that all six volunteers who consumed milk con-
taining 107 isolates of VRE produced stools containing
4 × 103–2 × 107 colony-forming units of VRE per gram of
stool after 1–2 days, and five of the six volunteers were still
colonized with VRE after six days.
Finally, Frei et al. [45] demonstrated that all Gram-positive
isolates (enterococci, lactobacilli and staphylococci) from
the crops and intestines of broilers were resistant to baci-
tracin (MIC > 256 µg/ml), even though their feed was free
of bacitracin. Bacitracin was, however, included in the feed
given to the parent animals. This implies that, under the
conditions prevailing, the broiler population must have
gained some of their intestinal microflora via contaminated
eggshells from their mothers.
References and recommended reading
Papers of particular interest, published within the annual period of review,
have been highlighted as:
• of special interest
•• of outstanding interest
1. Prescott JF: Antimicrobial drug resistance and its epidemiology.
• In Antimicrobial Therapy in Veterinary Medicine, edn 3. Edited by
Prescott JF, Baggot JD, Walker RD. Ames: Iowa State University
Press; 2000:27-49.
This paper is an excellent state-of-the-art discussion of the issue of antibiotic
resistance in the veterinary and medical fields in the latest edition of a
leading textbook.
2. Mazel D, Davies J: Antibiotic resistance in microbes. Cell Mol Life
• Sci 1999, 56:742-754.
This is a short general review that discusses integrons and their ancestry.
3. Levy SB: The Antibiotic Paradox: How Miracle Drugs are Destroying
the Miracle. New York: Plenum Press; 1992.
4. Committee on Drug Use in Food Animals, Panel on Animal Health,
• Food Safety, and Public Health, Board on Agriculture, National
Research Council, Food and Nutrition Board, Institute of Medicine:
The Use of Drugs in Food Animals: Benefits and Risks. Washington,
DC: National Academy Press; 1999.
This is an elaborate review of the literature and situation in the United States.
It concedes some problems but still plays down the impact and contribution
of antibiotics in animal production on ecological resistance spread. It is a
very useful source of detailed information on antibiotics approved for various
uses (therapy, prophylaxis, growth and feed efficiencies) in various animals
(pigs, cattle, sheep, chicken and turkeys).
5. Aarestrup FM, Bager F, Jensen MF, Madsen M, Meyling A,
Wegener HC: Resistance to antimicrobial agents used for
animal therapy in pathogenic, zoonotic and indicator
bacteria isolated from different food animals in Denmark:
a baseline study for the Danish Integrated Antimicrobial
Resistance Monitoring Programme (DANMAP). APMIS 1998,
106:745-770.
6. Teuber M: Spread of antibiotic resistance with food-borne
• pathogens. Cell Mol Life Sci 1999, 56:755-763.
This is a short review of the field of antibiotic resistance in food-borne
pathogens over the years 1995–1998. This paper documents that antibiotic
resistance genes invaded all major food-borne pathogens at different levels.
7. Charpentier E, Courvalin P: Antibiotic resistance in Listeria spp.
• Antimicrob Agents Chemother 1999, 43:2103-2108.
The authors of this paper compile data on antibiotic resistance in the
emerging food-borne pathogen, Listeria monocytogenes, and other rare Listeria
species. It points to the ubiquitous reservoir of Listeria species on, for example,
plants that are not normally under an antibiotic selection pressure.
8. Teuber M, Meile L, Schwarz F: Acquired antibiotic resistance in lactic
• acid bacteria from food. Antonie van Leeuwenhoek Int J 1999,
76:115-137.
This paper describes the acquisition of antibiotic resistance determinants by
lactic acid bacteria of human, animal and food origin. It demonstrates that
commensal enterococci, lactococci and lactobacilli in human and animal
bodies have picked up resistance genes with the aid of conjugative trans-
posons and plasmids that do contaminate food. The paper demonstrates
that raw sausages and raw milk cheeses contain antibiotic-resistant bacteria,
mainly enterococci, that come from the intestinal microflora of the
milk-producing animals. See also [34•,35,36•,42].
9. Schnabel EL, Jones AL: Distribution of tetracycline resistance
• genes and transposons among phylloplane bacteria in Michigan
apple orchards. Appl Environ Microbiol 1999, 65:4898-4907.
This is the first paper to be published on the distribution of tetracycline
resistance in plant pathogens and commensal bacteria.
10. Witte W: Medical consequences of antibiotic use in agriculture.
Science 1998, 279:996-997.
11. Molbak K, Baggesen DL, Aarestrup FM, Ebbesen JM, Engberg J,
•• Frydendahl K, Gerner-Schmidt P, Petersen AM, Wegener HC:
An outbreak of multidrug-resistant, quinolone-resistant
Salmonella enterica serotype Typhimurium DT104. New Engl J
Med 1999, 341:1420-1425.
This is one of the few studies that links pig Salmonella typhiumurium DT104
to a human outbreak resulting in two deaths.
12. Ungemach FR: Figures on quantities of antibacterials used for
•• different purposes in the EU-countries and interpretation. Acta Vet
Scand 2000, 93:89-98.
This is a thorough interpretation of available consumption data from Europe.
13. Harwood VJ, Whitlock J, Withington V: Classification of antibiotic
• resistance patterns of indicator bacteria by discriminant
analysis: use in predicting the source of fecal contamination in
subtropical waters. Appl Environ Microbiol 2000,
66:3698-3704.
The paper demonstrates clearly that antibiotic use has led to the development
of specific antibiotic resistance profiles in indicator bacteria of human and
animal origin that can be used for epidemiological classification and predictions
regarding environmental isolates. See also [14•].
14. Hagedorn C, Robinson SL, Filtz JR, Grubbs SM, Angier TA,
• Beneau RB: Determining sources of fecal pollution in a rural
Virginia watershed with antibiotic resistance patterns in fecal
streptococci. Appl Environ Microbiol 1999, 65:5522-5531.
See annotation to [13•].
15. Aminov RI, Garrigues-Jeanjean N, Mackie RI: Molecular ecology of
•• tetracycline resistance: development and validation of primers
for detection of tetracycline resistance genes encoding
ribosomal protection proteins. Appl Environ Microbiol 2001,
67:22-32.
This paper describes the development of nucleotide probes for all ribosomal
protection protein determinants responsible for tetracycline resistance, and
their application to distribution studies in swine and cattle experimental stations.
16. Chee-Sanford JC, Aminov RI, Krapac IJ, Garrigures-Jeanjean N,
• Mackie RI: Occurrence and diversity of tetracycline resistance
genes in lagoons and groundwater underlying two swine
production facilities. Appl Environ Microbiol 2001,
67:1494-1502.
This paper describes the application of tet resistance probes to monitor
swine effluents, lagoons and connected ground water. It clearly demonstrates
the release of tetracycline resistance genes from farm effluents into the soil
and ground water of the surrounding environment.
17. Smalla K, Heuer H, Götz A, Niemeyer D, Krögerrecklenfort E, Tietze E:
• Exogenous isolation of antibiotic resistance plasmids from
piggery manure slurries reveals a high prevalence and diversity
of IncQ-like plasmids. Appl Environ Microbiol 2000,
66:4854-4862.
This paper provides a clear molecular characterization of the presence and
nature of IncQ resistance plasmids in swine slurries.
18. Sundin GW: Examination of base pair variants of the strA-strB
streptomycin resistance genes from bacterial pathogens of
humans, animals and plants. J Antimicrob Chemother 2000,
46:848-849.

19. Zhao S, White DG, Ge B, Ayers S, Friedman S, English L, Wagner D,
• Gaines S, Meng J: Identification and characterization of integron-
mediated antibiotic resistance among shiga toxin-producing
Escherichia coli isolates. Appl Environ Microbiol 2001, 67:1558-1564.
The authors of this paper identify mechanisms of resistance and resistance
determinants in the medically important STEC, whose primary habitat is in cattle.
20. Guerra B, Soto SM, Argüelles JM, Mendoza MC: Multidrug
resistance is mediated by large plasmids carrying a class 1
integron in the emergent Salmonella enterica serotype (4,5,12:8:-).
Antimicrob Agents Chemother 2001, 45:1305-1308.
21. Daly M, Fanning S: Characterization and chromosomal mapping of
antimicrobial resistance genes in Salmonella enterica serotype
Typhimurium. Appl Environ Microbiol 2000, 66:4842-4848.
22. Ng LK, Mulvey MR, Martin I, Peters GA, Johnson W: Genetic
• characterization of antimicrobial resistance in Canadian isolates
of Salmonella serovar Typhimurium DT104. Antimicrob Agents
Chemother 1999, 43:3018-3021.
This is an interesting study on the molecular diversity of resistance determinants
in DT104.
23. Fey PD, Safranek TJ, Rupp ME, Dunne EF, Ribot E, Iwen PC,
•• Bradford PA, Angulo FJ, Hinrichs SH: Ceftriaxone-resistant
Salmonella infection acquired by a child from cattle. New Engl J
Med 2000, 342:1242-1249.
This is one of the few reports that directly links the source of infection
with a diseased patient. It also describes the emergence of a new
resistance determinant.
24. Dunne EF, Fey PD, Kludt P, Reporter R, Mostashari F, Shillam P,
• Wicklund J, Miller C, Holland B, Stamey K et al.: Emergence of
domestically acquired ceftriaxone-resistant Salmonella infections
associated with AmpC β-lactamase. JAMA 2000, 284:3151-3156.
The authors of this paper identify the mechanism of resistance of the newly
emerged cetriaxone-resistant Salmonella typhimurium.
25. Walker RA, Sauders N, Lawson AJ, Lindsay EA, Dassama M, Ward LR,
• Woodward MJ, Davies RH, Liebana E, Threlfall EJ: Use of a
lightcycler gyrA mutation assay for rapid identification of
mutations conferring decreased susceptibility to ciprofloxacin in
multiresistant Salmonella enterica serotype Typhimurium DT104
isolates. J Clin Microbiol 2001, 39:1443-1448.
This paper provides molecular proof that an already existing ACSSuT–
phenotype has developed independent chromosomal resistance to fluoro-
quinolones.
26. Carlson SA, Willson RM, Crane AJ, Ferris KE: Evaluation of
invasion-conferring genotypes and antibiotic-induced
hyperinvasive phenotypes in multiple antibiotic resistant
Salmonella typhimurium DT104. Microb Pathog 2000, 28:373-378.
27. Millemann Y, Gaubert S, Remy D, Colmin C: Evaluation of
• IS200-PCR and comparison with other molecular markers to trace
Salmonella enterica subsp. enterica serotype typhimurium bovine
isolates from farm to meat. J Clin Microbiol 2000, 38:2204-2209.
Modern automated PCR techniques prove that shedding of Salmonella from
carrier cows occurs all the way from the stable to the final meat product,
contaminating transport facilities, the slaughterhouse and other animals
along the way.
28. Saenz Y, Zarazaga M, Lantero M, Gastanares MJ, Baquero F,
•• Torres C: Antibiotic resistance in Campylobacter strains isolated
from animals, foods, and humans in Spain in 1997-1998.
Antimicrob Agents Chemother 2000, 44:267-271.
This is a worrying description of the increase of quinolone resistance from
zero in 1988 to almost 100% in 1998 in Camplylobacter isolates from
Spanish poultry and pigs, and from zero in 1988 to about 70% in 1997
and1998 human isolates.
29. Anderson SA, Yeaton Woo RW, Crawford LM: Risk assessment of
• the impact on human health of resistant Campylobacter jejuni from
fluoroquinolone use in beef cattle. Food Control 2001, 12:13-25.
This paper provides a description of a very cautious and conservative risk
assessment for humans of quinolone resistance development in beef.
30. Nawaz MS, Khan SA, Khan AA, Khambaty FM, Cerniglia CE:
Comparative molecular analysis of erythromycin-resistance
determinants in staphylococcal isolates of poultry and human
origin. Mol Cell Probes 2000, 14:311-319.
31. Khan SA, Nawaz MS, Khan AA, Cerniglia CE: Transfer of
erythromycin resistance from poultry to human clinical strains of
Staphylococcus aureus. J Clin Microbiol 2000, 38:1832-1838.
Veterinary use and antibiotic resistance Teuber 499
32. Haack BJ, Andrews RE: Isolation of Tn916-like conjugal elements
• from swine lot effluent. Can J Microbiol 2000, 46:542-549.
The authors of this paper demonstrate the wide distribution in swine effluents
of Tn916-like transposons from enterococci.
33. Kruse H, Johansen BK, Rorvik LM, Schaller G: The use of avoparcin
• as a growth promotor and the occurrence of vancomycin-resistant
Enterococcus species in the Norwegian poultry and swine
production. Microb Drug Res 1999, 5:135-139.
Vancomycin resistance in enterococci from Norwegian poultry houses is
disappearing very slowly, after a ban of avoparcin. See also [43•].
34. Giraffa G, Olivari A, Neviani E: Isolation of vancomycin-resistant
• Enterococcus faecium from Italian cheeses. Food Microbiol 2000,
17:671-677.
This paper describes the presence of VRE in many investigated Italian cheeses.
35. Baumgartner A, Kueffer M, Rohner P: Occurrence and antibiotic
resistance of enterococci in various ready-to-eat foods. Arch
Lebensmittelhyg 2001, 52:1-24.
36. Bertrand X, Mulin B, Viel JF, Thouverez M, Talon D: Common PFGE
• patterns in antibiotic-resistant Enterococcus faecalis from
humans and cheeses. Food Microbiol 2000, 17:543-551.
This paper shows that community-acquired high-level kanamycin- and gen-
tamicin-resistant enterococci may have their sources in certain cheese types.
37. Eaton TJ, Gasson MJ: Molecular screening of Enterococcus
•• virulence determinants and potential for genetic exchange
between food and medical isolates. Appl Environ Microbiol 2001,
67:1628-1635.
This is the first clear molecular demonstration of all eight investigated virulence
determinant classes in Enterococcus faecalis isolated from medical and
food sources.
38. Perreten V, Schwarz FV, Teuber M, Levy SB: Mdt(A), a new efflux
protein conferring multiple antibiotic resistance in Lactococcus
lactis and Escherichia coli. Antimicrob Agents Chemother 2001,
45:1109-1114.
39. Rhodes G, Huys G, Swings J, McGann P, Hiney M, Smith P,
• Pickup RW: Distribution of oxytetracycline resistance plasmids
between aeromonads in hospital and aquaculture environments:
Implication of Tn1721 in dissemination of the tetracycline
resistance determinant Tet A. Appl Environ Microbiol 2000,
66:3883-3890.
The authors of this paper stress and prove that human and aquaculture
compartments are not separated; instead, they are shown to contain the
same tet determinant and transposons.
40. Sorum H: Farming of Atlantic salmon — an experience from
•• Norway. Acta Vet Scand 2000, 93:129-134.
This paper provides a convincing example that proper vaccination against
pathogenic vibrios and aeromonads reduces antibiotic application in Atlantic
salmon aquaculture to nearly zero.
41. Teuber M: Antibiotikaresistente Bakterien in Lebensmitteln. Mitt
Lebensm Hyg 2001, 92:10-27. [Title translation: Antibiotic-resistant
bacteria in food.]
42. Schwarz FV: Antibiotic-resistant enterococci in food: complete
nucleotide sequence of the 50kbp conjugative multiresistance
plasmid pRE25 [PhD Thesis]. Zürich: Eidgenössische Technische
Hochschule Zürich; 2001. [Dissertation no. 14162. In October 2001,
the complete dissertation text will be available on the URL
http://e-collection.ethbib.ethz.ch/ediss/sg/10x.html]
43. Borgen K, Sorum M, Wasteson Y, Kruse H: VanA-type
• vancomycin-resistant enterococci (VRE) remain prevalent in
poultry carcasses 3 years after avoparcin was banned. Int J Food
Microbiol 2001, 64:89-94.
See annotation to [33•].
44. Aarestrup FM, Seyfarth AM, Emborg H-D, Pedersen K,
•• Hendriksen RS, Bager F: Effect of abolishment of the use of
antimicrobial agents for growth promotion on occurrence of
antimicrobial resistance in fecal enterococci from food animals in
Denmark. Antimicrob Agents Chemother 2001, 45:2054-2059.
A convincing study based on previous elaborate baseline data (see also [5]
and Update).
45. Frei A, Goldenberger D, Teuber M: Antimicrobial susceptibility of
intestinal bacteria from Swiss poultry flocks before the ban of
antimicrobial growth promoters. Syst Appl Microbiol 2001,
24:116-121.