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Pharmacokinetics of
Enrofloxacin in Reptiles
Mark G. Papich, DVM, MS, Diplomate ACVCP
Associate Professor of Clinical Pharmacology, College of Veterinary Medicine, North Carolina State University, Raleigh,
North Carolina, USA
Several fluoroquinolones are now
approved for use in veterinary medi-
cine. In North America, these include
enrofloxacin (Baytril® [Bayer]), orbi-
floxacin (Orbax™ [Schering-Plough
Animal Health]), difloxacin (Dicural®
[Fort Dodge Animal Health]), and mar-
bofloxacin (Zeniquin® [Pfizer Animal
Health]). Other fluoroquinolones have
also been used in small animal medi-
cine, such as the human drug cipro-
floxacin (Cipro® [Bayer]). Among these
drugs, enrofloxacin has received the
most attention for its use in exotic ani-
mals, especially in reptiles, and birds.
The fluoroquinolones have chemical
and biological properties that make it
ideal for use in reptiles; this paper
reviews these applications and reports
on the pharmacokinetic studies that
have been conducted with investiga-
tors from the University of Florida, the
Bronx Zoo, and the National Zoo of the
Smithsonian Institution in collaboration
with our laboratory. These investiga-
tions have examined the pharmaco-
kinetics of enrofloxacin in alligators,
snakes, tortoises, turtles, and lizards.
Pharmacologic Properties
of Enrofloxacin
Enrofloxacin is highly lipophilic, as
exhibited by its high octanyl:water par-
tition coefficient, and it exists as a zwit-
terion at most pH ranges. Studies con-
ducted in our laboratory have shown
that enrofloxacin is remarkably stable
when admixed with water, saline solu-
110 Third International Veterinary Symposium on Fluoroquinolones Proceedings
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tion, and various drugs. Because of its
durable chemical properties, enro-
floxacin in tablets or injectable solution
has been reformulated in various ways
to produce a convenient oral dosage
formulation. Because of its high lipid
solubility, enrofloxacin is highly ab-
sorbed after both intramuscular injec-
tion and oral administration, as our
studies show.
Pharmacokinetic
Characteristics
Enrofloxacin is well absorbed and
highly distributed in nearly all examined
animals. Oral absorption ranges from
40% in horses to nearly 100% in dogs
and cats. In species in which compar-
isons are available, enrofloxacin is
absorbed to a higher extent than is
ciprofloxacin. Enrofloxacin’s volume of
distribution ranges from 2 to 4 L/kg in
most species studied, demonstrating the
ability to penetrate intracellularly and to
attain high concentrations in tissues. In
most species in which elimination has
been examined, enrofloxacin shows
variable metabolism. In dogs, cats, adult
horses, cattle, and some reptiles, enro-
floxacin is metabolized partially to the
active metabolite ciprofloxacin. In other
animals, such as pigs, foals, lizards, and
crocodiles, little or no ciprofloxacin is
detectable in the plasma after enro-
floxacin administration.
Drug Analysis
Studies cited in this paper used a
specific and sensitive high-pressure liq-
uid chromatography (HPLC) method
similar to that reported in previous
work from our laboratory.1 The only
modification is that in some studies,
solid-phase extraction was used instead
of liquid extraction to provide better
sample cleanup. This assay is capable
of separating enrofloxacin from
ciprofloxacin and has an approximate
limit of detection of 0.02 µg/ml and a
limit of quantification of 0.05 µg/ml.
An assay that can distinguish between
these two active drugs is critical for
assessing pharmacokinetic characteris-
tics. A nonspecific assay, such as a
microbiologic determination, may pro-
duce erroneous data on the absolute
plasma concentrations of enrofloxacin.
Spectrum of Activity
The organisms most often causing
infection in reptiles include both gram-
positive and gram-negative bacteria,2 but
it has been suggested that gram-negative
bacteria are responsible for the most
important bacterial problems in reptiles.3
The gram-negative bacteria most often
implicated are Salmonella spp., (especial-
ly Salmonella arizonae), Serratia spp.,
Aeromonas spp., (especially Aeromonas
hydrophila), Klebsiella spp., Providencia
spp., and Pseudomonas spp. (especially
Pseudomonas aeruginosa). P. aeruginosa
is particularly important, because it is a
common pathogen, survives well in
aquatic environments, and can become
extremely resistant to many antibiotics,
even the fluoroquinolones. Many of
these bacteria can be part of a reptile’s
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normal flora of the oral cavity or gas-
trointestinal tract. However, even those
that are part of the normal flora can
become opportunistic invaders under
appropriate conditions and, if cultured
during an illness, should be considered
for their pathogenic potential.
The fluoroquinolones are active
against most of the gram-negative bac-
teria that affect reptiles, which makes
this class of drugs an ideal choice for
therapy. In addition, as summarized by
Jacobson,3 drugs such as the aminogly-
cosides that are highly active against
these gram-negative bacteria may have
long half-lives, increasing the risk of
nephrotoxicosis.3,4 In contrast, even though
fluoroquinolones have long half-lives in
many reptiles, there are no published
accounts of adverse effects when admin-
istered at the recommended therapeutic
dose rates. Therefore, fluoroquinolones
have the advantage of long half-lives
(which allows for infrequent dosing),
potent bactericidal activity, a good safety
record, and high activity against the bac-
teria that appear to cause the most prob-
lems in reptiles.
Pharmacokinetic Studies
Early reports of pharmacokinetic
studies of enrofloxacin in reptiles indi-
cated that further work was needed to
characterize its disposition in a variety
of species. Even though long half-lives
have been reported and translated to
long dosing intervals of 24 to 48 hours,
there is enough variation among
species to justify further work.2,3
Because investigators have encouraged
species-specific pharmacokinetic stud-
ies to learn more about the disposition
of fluoroquinolones in reptiles and to
devise dosage regimens, this paper
briefly summarizes some of the studies
conducted in collaboration with our
laboratory.
Alligators
Enrofloxacin was administered
intravenously (IV) to five alligators
(weight range 2.8–5.8 kg) at a dose of
5 mg/kg and orally to five alligators
(weight range 2.1–9.7 kg) at a dose of
5 mg/kg via a feeding tube.5 The alliga-
tors were kept at a temperature of 27˚C
throughout the study. After blood col-
lection for 96 hours, plasma was ana-
lyzed for enrofloxacin and ciprofloxacin
using an HPLC method mentioned pre-
viously. Ciprofloxacin was detected, but
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The
fluoroquinolones
are active
against most of
the gram-
negative
bacteria that
affect reptiles,
which makes this
class of drugs an
ideal choice for
therapy.
concentrations were low (0.09 µg/ml at
24 hours; peak plasma concentration
[Cmax] of 0.15 µg/ml at 30 hours) and
not reported in the table. After IV
administration, the mean half-life of
enrofloxacin was 55 hours and the vol-
ume of distribution was 3.75 L/kg.
After oral administration, the enro-
floxacin Cmax was only 0.45 µg/ml at 24
hours. The half-life after oral adminis-
tration, 120 hours, should be evaluated
cautiously, because a reliable terminal
half-life could be calculated in only two
of the animals (half-life in those ani-
mals averaged 77.5 hours). In the oth-
er animals, there was a long terminal
slope, and samples were not collected
long enough to obtain a true estimate
of the terminal half-life. In these ani-
mals, oral absorption obviously was
prolonged and influenced the rate of
elimination. The calculated percentage
of bioavailability (%F) of 169% after
oral administration in these animals
cannot be considered accurate because
of the inability to estimate the true ter-
minal slope of the curve in most ani-
mals.
This study shows that enrofloxacin
administered IV has a long half-life and
that oral administration seems to pro-
long the elimination even longer. Dos-
ing recommendations from these stud-
ies should be made cautiously
(discussed further at the end of this
paper). After IV injection, plasma con-
centrations attained an average maxi-
mum plasma concentration (Cmax, also
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called the “peak”) greater than 4
µg/ml, although considerable variabili-
ty in concentrations was noted. Using
a ratio of the Cmax to the minimum
inhibitory concentration (Cmax:MIC
ratio) that some have recommended
for determining dose rates, the IV dose
would be adequate to treat organisms
with an MIC of 0.25 to 0.5 µg/ml.
Using another dosing guideline of area
under the curve to the MIC ratio
(AUC:MIC ratio) greater than 125,
both the IV and the oral dose used in
this study would be adequate for treat-
ing bacteria with MIC less than 1.0
µg/ml using a dosing interval of 96
hours.
Savanna Monitors
Enrofloxacin was administered to 10
Savanna monitors at a dose of 10
mg/kg orally and intramuscularly (IM)
and to 7 Savanna monitors at a dose of
5 mg/kg orally and IM.6 The IM dose
was injected in the forelimb; the oral
dose was administered by feeding mice
that had been injected with enro-
floxacin. The monitors were kept at a
temperature of 27˚C throughout the
study. Blood samples were collected at
regular intervals and plasma assayed
for enrofloxacin and ciprofloxacin as
described earlier. Because of the moni-
tors’ small size and the stress and diffi-
culty in obtaining multiple blood sam-
ples from each animal, the number of
blood samples was limited. Therefore,
five of the animals in the 10 mg/kg
group (both routes) were sampled at
0.5, 3, 12, and 36 hours, and the oth-
er five animals were sampled at 1, 6,
and 24 hours. To ensure that samples
were collected for a sufficient interval,
at another date an additional dose was
administered to three animals and sam-
ples collected at 48, 72, and 96 hours
for each route. The concentrations for
each time point were averaged and
pharmacokinetic parameters calculated
from the mean values. For the 5 mg/kg
dose, four animals were administered
an IM dose and three animals were
administered an oral dose; all were
sampled at 6, 24, 48, 72, and 96
hours.
The terminal elimination half-life
was prolonged for both routes and
both dose rates. The terminal half-life
was 40 hours from both routes after a
dose of 5 mg/kg, and 36 and 24 hours
for IM and oral doses, respectively, at
10 mg/kg. The Cmax was high for both
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Figure 1—Mean enrofloxacin and its metabolite ciprofloxacin
concentrations after enrofloxacin administration to Burmese
pythons at a dose of 5 mg/kg IM (n = 6).
dose rates, 1.99 µg/ml for both routes
after 5 mg/kg, and 10.5 µg/ml and 3.6
µg/ml for IM and oral doses, respec-
tively, at 10 mg/kg. The high peak and
long elimination rate resulted in total
AUC values that ranged from 118
µg/hour/ml after 5 mg/kg IM to 456
µg/hour/ml after 10 mg/kg IM.
The Cmax for enrofloxacin was high
for both dose rates in comparison to
similar doses in other species. There
was some metabolism of enrofloxacin
to ciprofloxacin, but we did not report
on the pharmacokinetics in the tables
because the concentrations were too
low to obtain meaningful estimates.
Ciprofloxacin concentrations after IM
administration were undetectable in
the early samples at a dose of 10
mg/kg and less than 0.1 µg/ml for most
samples in which it was detectable.
Concentrations were below the limit of
quantification (LOQ) in most samples
after oral administration at 10 mg/kg.
After the dose of 5 mg/kg, cipro-
floxacin concentrations were below the
LOQ in most samples.
Using dosing criteria stated earlier,
the lower dose administered IM or orally
would be adequate to treat moderately
susceptible bacteria such as Entero-
bacteriaceae. The higher dose of 10
mg/kg produced high peaks that may
be adequate, judging from the Cmax:MIC
or AUC:MIC ratio, to treat bacteria with
high MIC values, such as Pseudomonas.
For each dose, the long half-life sug-
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Figure 2—Mean enrofloxacin and its metabolite ciprofloxacin
concentrations after enrofloxacin administration to Burmese
pythons at a dose of 5 mg/kg IM once daily for 5 days (n = 5).
gests that once every 96 hours would
be adequate if one uses the criteria of
AUC:MIC for dose determination.
Burmese Pythons
Enrofloxacin was administered to 11
Burmese pythons (weight range
0.75–1.75 kg) that were kept at 26˚C
throughout the study.7 Six of the
pythons received a single IM dose of 5
mg/kg, and blood samples were col-
lected at regular intervals for 96 hours.
Enrofloxacin was administered IM to
the remaining five pythons at a dose of
5 mg/kg once daily for 5 days. During
the multiple-dose study, blood samples
were collected immediately before
each injection (trough) and 6 hours
after the injection (peak). Plasma from
the blood samples was harvested for
enrofloxacin and ciprofloxacin HPLC
analysis.
A considerable amount of cipro-
floxacin was metabolized from
enrofloxacin in both the single- and
multiple-dose studies. After the single
dose, the mean terminal half-life for
enrofloxacin was 6.37 hours. The peak
concentrations were 1.66 µg/ml and
0.35 µg/ml for enrofloxacin and cipro-
floxacin, respectively. As shown in Fig-
ure 1, ciprofloxacin appeared to persist
longer than enrofloxacin, even though
samples were not collected beyond 96
hours.
After the multiple-dose study, the
mean half-life, measured from an esti-
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mate of the peak–trough slopes after
each dose, was longer for every dose
(Figure 2), compared to the single-dose
study. With once daily dosing at 5
mg/kg, both enrofloxacin and
ciprofloxacin appeared to accumulate.
Results from the single-dose study sug-
gest that 5 mg/kg would be effective
for organisms with MICs ≤0.2 µg/ml (as
estimated from Cmax:MIC ratios or
AUC:MIC ratios). However, because
there were higher Cmax and AUC from
accumulation with repeated doses, 5
mg/kg administered every 48 hours
would produce adequate plasma con-
centrations for sensitive bacteria, such
as Escherichia coli. But because P.
aeruginosa often has higher MIC val-
ues (0.5 [±0.25] µg/ml from data col-
lected for this study), higher doses
should be considered for this organism.
Indian Star Tortoises
Enrofloxacin was administered to 92
Indian star tortoises.8 Another 20 tor-
toises served as controls. Their weights
ranged from 204 to 713 g. Enro-
floxacin was administered IM in the
front leg at a dose of 5 mg/kg, and
blood was collected for plasma analysis
of enrofloxacin and ciprofloxacin using
an HPLC technique. Because of the ani-
mal’s small size, only one blood sample
was collected from each tortoise. The
animals were therefore assigned to
sampling groups, with 6 to 15 tortoises
per group, and pooled data were used
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Figure 3—Mean enrofloxacin and its metabolite ciprofloxacin concentrations after
enrofloxacin administration to Indian star tortoises at a dose of 5 mg/kg orally (n = 92).
for pharmacokinetic analysis. The times
for each sampling group were 0, 0.5,
1, 1.5, 2, 4, 12, 24, 48, 72, and 96
hours after drug administration. The
tortoises were kept at a temperature of
26˚ to 30˚C throughout the study.
Concentrations and plasma versus
time curves after enrofloxacin adminis-
tration are shown in Figure 3. A signifi-
cant amount of ciprofloxacin was
metabolized from enrofloxacin, which
differs from data in other published
papers on tortoises.9 In this study, the
terminal half-life of enrofloxacin and
ciprofloxacin was 5.1 and 5.8 hours,
respectively, which is considerably short-
er than the half-life in gopher tortoises9;
however, the gopher tortoises weighed
approximately 10 times more than the
Indian star tortoises used in this study.
The mean Cmax was 3.59 and 0.35 for
enrofloxacin and ciprofloxacin, respec-
tively. The ciprofloxacin concentrations,
although low, would be expected to
contribute to the antibacterial effect
from enrofloxacin administration. Data
from this study suggest that this dose of
5 mg/kg q24h would be adequate to
treat infections caused by bacteria with
MICs of approximately 0.5 µg/ml and
lower, using a Cmax:MIC ratio of 8 to 10,
as some have suggested.
Red-Eared Sliders
Enrofloxacin was administered to six
red-eared sliders at a dose of 5 mg/kg
orally and IM, with a washout period
between oral and IM studies.10 After
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drug administration by each route,
blood samples were collected at 0.5, 1,
2, 4, 8, 12, 24, 36, and 48 hours, and
plasma was analyzed for enrofloxacin
and ciprofloxacin by HPLC analysis.
Enrofloxacin appeared to be well
absorbed from each route, with a
mean Cmax of 6.28 and 3.45 µg/ml after
IM and oral dosing, respectively. The
absolute amount absorbed is not
known because there was no accom-
panying IV study, but the ratio of AUC
from oral and IM (AUCoral:AUCIM)
administration was 0.94. Despite the
apparent high amount of absorption
from the oral dose, this route showed a
delayed time to peak (Tmax) and a longer
elimination half-life (IM half-life and
oral half-life were 17.6 and 32.8 hours,
respectively). There was considerable
ciprofloxacin metabolism from enro-
floxacin regardless of route, with mean
Studies
reported here
showed that
enrofloxacin is
well absorbed
from IM
injection in
reptiles.
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ciprofloxacin Cmax values of 0.36 and
0.29 µg/ml after IM and oral doses,
respectively. Ciprofloxacin is expected
to contribute to antibacterial effects
because it is an active drug.
The dose of 5 mg/kg used here,
either orally or IM, would be expected
to produce antibacterial effects for
most gram-negative bacteria with an
MIC ≤0.5 µg/ml after a single dose
(using a suggested Cmax:MIC ratio of 8
to 10). The frequency of this dose can-
not be determined without further
study, but the long half-life implies that
long dosing intervals could be used. In
these red-eared sliders, the AUC:MIC
ratio was above 100 to 125 for bacte-
ria with MIC ≤0.5 µg/ml for approxi-
mately 48 hours after an IM dose and
for 72 hours after an oral dose. Accord-
ing to some guidelines, these intervals
may be adequate for a bacterial cure.
Dosing Strategies
Selection of fluoroquinolone doses
for mammals has focused on adminis-
tration of a sufficient amount to attain
either an optimum Cmax:MIC ratio or
AUC:MIC ratio. The magnitude of
these ratios has been debated. In mam-
mals, a Cmax:MIC ratio of at least 8 to
10 or an AUC:MIC ratio of at least 125
has been suggested.11 However,
because these ratios were obtained
from studies in animals that were
immunocompromised or in critically ill
humans, they may be higher than that
needed in immunocompetent animals.
Also, the AUC:MIC ratio used in mam-
mals is based on an AUC for a 24-hour
interval. Because enrofloxacin is admin-
istered in many reptiles every 48 to 72
hours or longer, we do not know if the
AUC should include a longer interval.
Despite these uncertainties, we need a
starting point. Using average MIC val-
ues for sensitive E. coli and P. aerugi-
nosa, we can begin to make some dos-
ing estimates, which have been
discussed earlier in the paper. However,
we should not determine doses on
pharmacokinetics alone. The next step
should be to test the efficacy of these
dosing estimates in clinical studies.
Routes of Administration
Studies reported here showed that
enrofloxacin is well absorbed from IM
injection in reptiles. The IM route of
administration is commonly used and
would be appropriate for enrofloxacin
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as long as it does not induce tissue
injury at the injection site. In studies
reported in this paper, the small animal
formulation of enrofloxacin (Baytril®)
was used (22.7 mg/ml). It is not known
if formulations of enrofloxacin that
contain other vehicles, or that are pre-
pared in a more concentrated solution,
will be absorbed to the same extent, or
produce tissue injury that is different
from the formulation used in this study.
For other drugs administered IM in
mammals, it has been observed that a
more concentrated formulation inject-
ed IM has considerably altered the
absorption characteristics of the drug.
The optimum site for IM injections
has not been compared for fluoro-
quinolones in reptiles. Reptiles have a
renal portal system (RPS) that can poten-
tially affect drug clearance. The RPS
shunts blood from an injection in the
rear legs to the kidneys before it enters
the circulation systemically. This may
decrease systemic availability of some
drugs. An RPS effect on the clearance of
enrofloxacin has not been investigated.
In a study with gentamicin,12 we
showed that the RPS had no influence
on disposition of gentamicin injected in
the rear legs of turtles. Presumably, the
RPS would have an effect only on drugs
for which there is a high first-pass effect
and rapid clearance by the kidneys. This
is doubtful for enrofloxacin because it
exhibited such a long half-life in the
studies reported here.
The oral route prolonged the
absorption in the species studied, as
shown by a long time to Tmax or a long
terminal half-life. Gastrointestinal tran-
sit time and oral absorption of anti-
biotics can vary among reptiles,
depending on ambient temperature
and feeding habits. Other investigators
have questioned reliance on the oral
route of administration in sick reptiles
114 Third International Veterinary Symposium on Fluoroquinolones Proceedings
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Temperature 3.
has been shown
to have an 4.
effect on the
pharmacokinetics 5.
of some drugs
in reptiles.
6.
because of the unpredictability of oral
absorption.3 Although the studies
reported here were performed on
healthy animals only, the data suggest 7.
that the oral route of administration for
enrofloxacin could be considered for
some conditions in reptiles.
Temperature has been shown to
have an effect on the pharmacokinetics 8.
of some drugs in reptiles. In these stud-
ies, effect of changing temperature
was not examined because all animals
were kept at a constant temperature. It
9.
is usually recommended that reptiles be
treated at the high end of the optimum
temperature range because adequate
drug clearance is ensured at high tem-
10.
peratures and bacterial infections
appear to respond better when animals
are kept at higher temperatures.2
11.
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