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Efficacy of toltrazuril as a metaphylactic and therapeutic treatment of

coccidiosis in first–year grazing calves

Article  in  Parasitology Research · November 2005

DOI: 10.1007/s00436-005-1456-x · Source: PubMed

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Parasitol Res (2005) 97:S127–S133
DOI 10.1007/s00436-005-1456-x

Efficacy of toltrazuril as a metaphylactic and

therapeutic treatment of coccidiosis in first-year
grazing calves

C. Epe1 (✉), G. von Samson-Himmelstjerna1, N. Wirtherle1, V. von der Heyden1, C. Welz1,

J. Beening3, I. Radeloff2, K. Hellmann2, T. Schnieder1, K. Krieger4
1Institute
of Parasitology, Hannover School of Veterinary Medicine, Buenteweg 17, 30559 Hannover, Germany
2Klifovet
AG, Geyerspergerstrasse 27, 80689 München, Germany
3Animal Clinic Grosstierpraxis Ahrens-Nordholt, Hillrichs, Kanal 8, 26847 Detern, Germany
4Bayer HealthCare AG, Animal Health, R&D Parasiticides, 51368 Leverkusen, Germany

(✉) e-mail: [email protected]

Abstract the toltrazuril- compared to the placebo-treated ani-

mals. Body weight in the toltrazuril-treated animals
A multicentric, placebo-controlled, randomised, blind- significantly exceeded that of the placebo-treated ani-
ed and blocked field study was conducted to evaluate mals at the end of the observation period. Mean differ-
the efficacy and safety of toltrazuril (Baycox®, Bayer ence in body weight was higher in the metaphylactic
AG, Leverkusen, Germany) in the treatment of coc- (+7.3 kg) compared to the therapeutic treatment group
cidiosis in first-year grazing calves naturally infected (+3.4 kg). No adverse reactions were observed. The
with Eimeria spp. Three-hundred and thirty-one calves results indicate that toltrazuril is highly efficacious and
were enrolled in the study and allocated to one of two safe in the metaphylactic and therapeutic treatment of
treatments at a ratio of 1:1. One hundred and sixty- coccidiosis caused by E. alabamensis in first-year graz-
seven animals were treated once orally with 15 mg/kg ing calves.
toltrazuril, and 164 animals served as placebo-treated
controls. Two treatment regimes were compared, a
metaphylactic (treatment on the day, or 1 day after,
turn out) and a therapeutic treatment (4 or 7 days after Introduction
turn out). During an observation period of 14 days
after treatment the animals were clinically examined Coccidia of the genus Eimeria may cause watery to
for diarrhoea and faecal samples were regularly bloody diarrhoea in many vertebrates. In cattle, several
assessed for Eimeria oocysts. Other possible causes of Eimeria species have been shown to cause clinical dis-
diarrhoea were excluded on the basis of microbiologi- ease associated with diarrhoea, high morbidity and
cal and virological examination. Animals were pre- even mortality. In the field, usually infections with
dominantly infected with Eimeria alabamensis. Num- more than one Eimeria species are found, but also
ber of days with diarrhoea in animals treated with monoinfections may occur (Gräfner et al. 1982; Svens-
toltrazuril was significantly lower compared to the son et al. 1993). In various European investigations E.
placebo-treated group (therapeutic treatment: alabamensis was identified as the dominating species
P=0.0024; metaphylactic treatment: P<0.0001). Fur- in grazing cattle with clinical coccidiosis. Other species
thermore, the number of animals with diarrhoea dur- frequently found are E. bovis, E. auburnensis, E. zuernii,
ing the observation period for a minimum of at least 3 E. ellipsoidalis and E. bukidnonensis (Svensson et al.
days, the number of animals positive for Eimeria 1993, 1994; Svensson 1997). During clinical outbreaks
oocysts, and the number of animals with both diar- of eimeriosis in grazing cattle in the northern part of
rhoea for a period of at least 3 days and positive for the former German Democratic Republic (GDR)
Eimeria oocysts, were significantly lower (P<0.01) in (Gräfner et al. 1982), E. alabamensis was the dominat-
>>
S127
 ing species in ten out of 18 herds examined, and in Two different treatment regimes were used:
three herds a monoinfection with E. alabamensis was
diagnosed. Infections with E. bovis dominated in four 1. Metaphylactic treatment: treatment of calves at the
herds, while in one herd a mixed infection with the time of supposed infection (day of turn out or 1 day
three pathogenic species E. alabamensis, E. bovis and E. after turn out to pasture).
zuernii was found. This reflects the typical situation in
northern Germany and, as far as is known, in central 2. Therapeutic treatment: treatment of calves after the
and northern Europe (Svensson et al. 1994; Samson- expected onset of clinical symptoms of coccidiosis 4–7
Himmelstjerna et al., in review). Typically E. alabamen- days after turn out to pasture.
sis infections show a prepatent period ranging between
6 and 10 days. Clinical disease usually starts at 3–7 For each treatment regime animals were allocated to
days post-infection with symptoms including watery either a toltrazuril group or a placebo group (water/
diarrhoea, poor appetite, depression, abdominal pain milk application) at a 1:1 ratio, according to pre-estab-
and reduced growth rate leading to a significant reduc- lished per centre (farm) randomisation lists that were
tion in weight gain or, at a low rate, even mortality available to the dispenser only (Table 1).
(Hooshmand-Rad et al. 1994; Svensson et al. 1994).
The primary evaluation criterion for the efficacy of the
The objective of this study was to evaluate the clinical treatment was the number of days animals showed
and parasitological efficacy as well as the safety of clinical signs of coccidiosis (diarrhoea) as measured by
toltrazuril in the metaphylactic and therapeutic treat- faecal scores. The tested hypothesis was that both
ment of first-year grazing calves under field condi- metaphylactic and therapeutic treatment with
tions, using a single oral dose of 15 mg toltrazuril/kg toltrazuril lead to a significant reduction in clinical
bodyweight. Efficacy and safety parameters were com- signs of coccidiosis as compared to the respective con-
pared between animals receiving toltrazuril and place- trol group.
bo-treated animals.
Secondary evaluation criteria were the number of
Eimeria oocysts shed, the percentage of calves with
diarrhoea for at least a 3-day period within the obser-
Materials and methods vation period and being positive for Eimeria oocysts,
and weight gain during the observation period of 14
Design of the study days.

The study was designed as a multicentric, placebo-con-

trolled, randomised, blinded and blocked field study
conducted on 14 dairy farms located in north-west Study animals
Germany which had a history of clinical coccidiosis in
calves after first turn out to pasture. It was performed Three-hundred and thirty-one calves – all but one
in accordance with the European guidelines for the (Spotted) Holstein Fresians – were enrolled in the
conduct of clinical trials as described in the directives study and allocated to one of two treatment regimes. In
2001/82/EC, EC 92/18, and VICH GL9 (GCP) 15 June total, 53 male (25 in treatment, 28 in placebo groups)
2000, Good clinical practice, and the Guideline on sta- and 278 female (139 in treatment and 139 in placebo
tistical principles for veterinary clinical trials groups) calves with an age range of 2–15 months par-
(EMEA/CVMP/816/00-Final). Randomisation was ticipated in the study. Body weight at the start of the
based on the weight of the animals on the day of turn study ranged from 61 kg to 357.5 kg with no significant
out to pasture. difference between treatment and placebo group
means. None of the animals was kept on pasture before
the beginning of the trial. The number of animals
enrolled per farm ranged between eight and 40. All
herds were kept on pastures close to the farms which

S128
 upper limit the number of
100,000 opg was used for cal-
culation. Another faecal sam-
ple was collected before treat-
ment for bacteriological and
virological examination to
confirm the absence of Salmo-
nella spp. and/or coronavirus.
This was repeated for each
animal when it had diarrhoea
for the first time. The microbi-
ological examinations were
done according to the stan-
dard methods of the Institute
of Microbiology (Veterinary
School Hannover) for the iso-
lation and identification of
had been routinely used in the years before but had not Salmonella spp. and, if applicable, Clostridium perfrin-
been grazed in the current year. gens.Virological examinations were performed accord-
ing to the test description of the coronavirus Elisa test
kit (Bio-X Coronavirus Elisa kit; Bio-X diagnostics,
Marche-en-Famenne, Belgium).
Clinical examination

A physical examination was performed by the examin- Treatment allocation and treatment
ing veterinarian on the treatment day, followed by daily
clinical observation for a 14-day observation period Half of the animals on each pasture were treated with
with an obligatory evaluation of faecal consistency. toltrazuril and the remaining half were allocated to the
placebo-treated control group according to the ran-
domisation procedure. Where possible, the pasture was
split into two equal parts. Immediately following appli-
Body weight cation animals were separated with regard to treat-
ment. Toltrazuril was applied as a commercial oral sus-
All animals were weighed on the treatment day and at pension containing 50 mg active per ml (toltrazuril;
the end of the observation period of 14 days. BayerHealthCare, Leverkusen, Germany) at a dose rate
of 15 mg/kg body weight. The toltrazuril dose was cal-
culated based on the individual animal’s body weight
recorded on the same day.Animals allocated to the con-
Faecal samples trol group received the same dose volume, of an oral
placebo, as the toltrazuril-treated animals.
Faecal samples were collected from each animal before
turn out, on the day of treatment and at regular inter-
vals after treatment and examined for pathogenic
Eimeria oocysts (Eimeria alabamensis, E. bovis, E. Animal selection and identification
zuernii), using a modified McMaster technique.
Oocysts were identified to species level and counted, All animals were reared on commercial farms and
i.e. up to 2,000 oocysts per species and sample [100,000 identified by individual ear tags. Only healthy animals
oocytes per gram of faeces (opg) calculated]; the sur- that had not previously been treated with anticoccidi-
plus was disregarded and for all counts exceeding this als or corticosteroids were enrolled. >>
S129
 Animal management and housing Results
Animals were kept on pastures providing between 18 Calves treated with toltrazuril or the placebo were
m2 and 105 m2/individual animal. The use of addition- comparable with regard to age, gender, weight and
al medication and/or therapies, which could directly physical examination results prior to the onset of the
affect the performance of the treatment product or the study (data not shown).
control group, i.e. sulphonamides, was not permitted.
All animals had permanent access to water and on 12 of Diarrhoea was first observed 4 days after turn out. The
the 14 farms they received feed concentrates and on highest percentage of animals showing diarrhoea dur-
two farms maize silage or milk was fed additionally. ing the observation period was 90%, recorded for the
placebo-treated animals of the metaphylactic treat-
ment group. The toltrazuril-treated animals had signif-
icantly fewer days with diarrhoea than the placebo-
Statistical analysis treated calves, for both the therapeutic (P=0.0024) and
metaphylactic (P<0.0001) treatment regimes (Fig. 1).
All variables measured for animals during the study The percentage of animals with diarrhoea was consis-
were stratified by treatment group and compared tently lower throughout the observation period in
descriptively. For continuous variables, the following toltrazuril compared to placebo-treated animals for
descriptive statistics were calculated where applicable: both treatment regimes (Figs. 2, 3). During the observa-
sample size, arithmetic and geometric means, SD, tion period only E. alabamensis and E. bovis oocysts
median, minimum, maximum, 25% percentile, and were recorded. A marked increase in oocyst numbers
75% percentile. For animal categorical or binary vari- was observed in both placebo-treated groups and in
ables, absolute and relative frequencies were displayed. the group receiving toltrazuril as a therapeutic treat-
To assess treatment group comparability before treat- ment, with a maximum opg count at day 11 after turn
ment various animal characteristics were compared on out in both placebo-treated groups and on days 9–11 in
the treatment day (baseline). For the comparison of the the group treated therapeutically with toltrazuril. In
number of days where the calves in the different groups contrast, oocyst numbers remained low in the group
had clinical signs of coccidiosis, i.e. diarrhoea, an receiving toltrazuril as a metaphylactic treatment
analysis of covariance (ANCOVA) was carried out (Figs. 2, 3). The vast majority of oocysts belonged to E.
using age, sex, and body weight as covariates. All other alabamensis. The daily opg counts for E. bovis did not
comparisons were carried out by employing a logistic exceed 482 in the placebo-treated animals and 374 in
regression model, using covariates age, sex and base- the toltrazuril-treated animals.
line body weight. The comparison of oocytes per gram
of faeces between treatments was carried out using a The percentage of animals with diarrhoea for a period
repeated measures model for binary variables. Weight of at least 3 days and opg-positive for Eimeria spp. dur-
gain was analysed using an ANCOVA, using covariates ing the observation period was 54.7% and 58.0% for
age, sex and baseline body weight. For all statistical the therapeutic and metaphylactic treatment, respec-
tests a nominal significance level of 5% (a=0.05) was tively, in animals receiving placebo, and 31.3% and
used. Although multiple outcome variables were used 11.6%, respectively, for animals receiving toltrazuril. In
to evaluate efficacy, no explicit multiple comparison both the metaphylactic and the therapeutic treatment
correction of the P-values was used. regimes the number of animals with diarrhoea, the
number of animals with diarrhoea and positive for opg
The occurrence of adverse events was reported counts, and the number of animals with diarrhoea for
descriptively. at least a 3-day period, was significantly lower in the
toltrazuril treatment group compared to the placebo-
treated group (all P<0.01).

Body weight gain in toltrazuril-treated animals signifi-

cantly (P<0.01) exceeded that in placebo-treated ani-

S130
 4 8
3.5
days with diarrhoea

3
7
2.5 6
(mean)

2
1.5 5
1
4

kg
+ 7.3 kg
0.5
0 3
therapeutic treatment metaphylactic treatment
2
+ 3.4 kg
toltrazuril placebo
1

Fig. 1. Mean duration (days) of diarrhoea per group 0

therapeutic treatment metaphylactic treatment

21000 60 Fig. 4. Weight gain during observation period of toltrazuril-

vs. placebo-treated calves after therapeutic and metaphylactic
oocyst shedding (opg)

18000 50
treatment
% animals with
15000
40
12000 diarrhoea
30
9000
20
6000
3000 10

0 0
Clostridium perfringens based on clinical signs.
1 2 3 4 5 6 7 8 9 10 11 12 13 14
days after turnout
toltrazuril placebo No suspected adverse drug reaction, local or systemic,
was reported for any animal treated with toltrazuril.
Fig. 2. Shedding of Eimeria oocysts (columns) and percentage
of calves with diarrhoea (lines) after metaphylactic treatment
with toltrazuril vs. placebo

Discussion
12000 60
In central and northern Europe coccidiosis in grazing
oocyst shedding

10000 50
% animals with

herds during the grazing season has previously been

diarrhoea

8000 40
(opg)

6000 30 described (Gräfner 1989; Svensson et al. 1994; Snoep

4000 20
and Potters 2004). For northern Germany, reports exist
2000 10
0 0
which describe clinical outbreaks as well as the epi-
7 8 9 10 11 12 13 14 15 16 17 18 demiological situation in grazing cattle on a multi-herd
days after turnout basis (Gräfner 1989). The animals usually showed diar-
placebo toltrazuril rhoea and weight loss during the first weeks following
Fig. 3. Shedding of Eimeria oocysts (columns) and percentage
turn out and occasionally mortality was reported. The
of calves with diarrhoea (lines) after therapeutic treatment pattern of E. alabamensis infections in first-year graz-
with toltrazuril vs. placebo ing calves is characterized by significant increases in
oocyst counts starting at approximately 7–9 days after
turn out (Snoep and Potters 2004). Infections are con-
sidered to be mainly caused by oocysts which persisted
mals. Metaphylactic treatment with toltrazuril resulted on the pasture since the previous grazing season
in a higher body weight gain during the observation (Gräfner et al. 1982; Svensson et al. 1993, 1994). The
period (+7.3 kg) compared to placebo-treated animals findings of the present study are consistent with this
than therapeutic treatment (+3.4 kg) (Fig. 4). view. In all herds and in nearly all of the calves E. alaba-
mensis oocysts were found and in approximately 80%
During the observation period no animal showed an of the animals clinical coccidiosis was diagnosed. The
infection with either Salmonella spp. or coronavirus pathogenic Eimeria species encountered in the present
and no animal was suspected to be infected with study were E. alabamensis and E. bovis, which were also >>
S131
 the most prevalent in an earlier study in the GDR when fed to calves in milk starter.Although the applica-
(Gräfner et al. 1982). Although infection with this tion and efficacy of this compound is well described
pathogen is usually considered to be acquired during (Hoblet et al. 1989; Sinks et al. 1992; Waggoner et al.
the first few days of grazing (Gräfner et al. 1982; Svens- 1994; McMeniman and Elliott 1995; Quigley et al. 1997),
son et al. 1993), it is also known that calves may be a major constraint is the labour-intensive step of mix-
infected before turn out, usually leading to the excre- ing the pure compound into the starter. Even though it
tion of low numbers of oocysts. It has been shown that is available as a commercial product (Bovatec, Hoff-
E. alabamensis oocysts can be transmitted by contami- man-La-Roche), farmers have had to be persuaded to
nated hay (Svensson 1997). On several of the farms use it; however, once applied, the compliance rate
included in this study, anecdotal reports of calves reported was very good (unpublished observations).
which died with signs of clinical coccidiosis during the Further coccidiostatics such as decoquinate (Rhone-
first 2 months following turn out were given. In con- Poulenc) or Monensin, known to be effective against
trast to E. bovis which can cause significant lesions in clinical coccidiosis in prophylactic use (Fitzgerald and
the large intestine with morbidity and mortality Mansfield 1984; Fitzgerald and Mansfield 1989a, 1989b;
(Daugschies et al. 1997; Taylor and Catchpole 1994), E. Foreyt et al., 1986; Stromberg et al. 1986; Waggoner et
alabamensis is known to cause clinical cases with lower al. 1994) are no longer registered for use in livestock
morbidity (Hooshmand-Rad et al. 1994). Nevertheless, animals in the European Union.
such natural infections usually contribute to reduced
weight gain during the first months of grazing and may Immunisation has also been presented as a possible
occasionally require treatment to avoid a fatal outcome option for prevention in preliminary investigations.
of the disease. However, this has not yet become commercially avail-
able.
To date, in many countries the prophylactic and cura-
tive options for the treatment of calves infected with The present study showed that treatment with
Eimeria spp. are unsatisfactory. For example, in Ger- toltrazuril (Baycox), applied according to a therapeutic
many, currently only sulphonamides are registered for or metaphylactic treatment regime, was safe and high-
the treatment of bovine coccidiosis. However, repeated ly efficacious against infections with Eimeria spp. in
applications of these drugs for several days only result first-year grazing calves. Treatment with toltrazuril sig-
in the reduction of clinical symptoms but are unable to nificantly decreased the number of days animals had
prevent these effectively. Experience with sulphon- diarrhoea. Furthermore, the number of animals with
amide treatment against bovine coccidiosis has been diarrhoea and/or the number of animals positive for
described worldwide. A sulphonamide-bolus-based Eimeria spp. was significantly lower in animals treated
treatment using a baquiloprim/sulphadimidine bolus with toltrazuril compared to the placebo-treated ani-
has been studied in Sweden and found to have signifi- mals, and treatment with toltrazuril resulted in a sig-
cant effects on reduced weight loss and oocyst output nificantly higher body weight compared to placebo-
in treated calves (Svensson 1998). Nevertheless, it is not treated animals.
marketed as a registered product and may be used in
niche markets only due to the handling expenses. Other The results obtained are in accordance with efficacy
combinations of molecules tested for enhanced effica- data for toltrazuril from an experimental E. bovis infec-
cy, such as sulphamonomethoxine/ormetoprim (Has- tion model with toltrazuril (Mundt et al. 2003) and
bullah et al. 1996), are reported to be effective as well. from a natural coccidiosis outbreak in calves treated
However, a multitude of reports concerning the unsat- with toltrazuril (Bohrmann 1991). However, possible
isfactory efficacy of these combinations in the field differences in formulation have to be considered since
reflect the decreased compliance of users for this class the off-label use of this compound in the 1990s did not
of compounds. employ the formulation used in the present study. Nev-
ertheless, in both publications a significant reduction
Among other coccidiostatics used for prophylactic in oocyst shedding, as well as an improvement of clini-
treatment, lasalocid has been known for many years to cal symptoms, were observed. Furthermore, in the
be effective in the prevention of clinical coccidiosis, experimental infection model against E. bovis the com-

S132
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