Veterinary Parasitology 129 (2005) 179–186

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Responses of chickens vaccinated with a live attenuated

multi-valent ionophore-tolerant Eimeria vaccine
G.Q. Li *, S. Kanu, S.M. Xiao, F.Y. Xiang
Department of Veterinary Parasitology, College of Veterinary Medicine, South China Agricultural University,
Guangzhou 510642, PR China
Received 11 June 2004; received in revised form 25 August 2004; accepted 7 September 2004

Abstract

Coccidiosis, caused by Eimeria species, is a serious economic disease of chickens (Gallus gallus) and the search for vaccines
to control the disease is intensifying especially with the increasing threat of drug resistance. A live attenuated multi-valent
ionophore-tolerant Eimeria vaccine has been developed that contains three ionophore-resistant Eimeria species, E. tenella, E.
maxima and E. acervulina. The attenuated lines were derived from virulent field strains resistant to monensin ionophore by
selection for early development in chicks. The vaccine was administered by gavage and through drinking water to broiler
chickens, Chinese Yellow strain, reared in wire cages. Vaccinated medicated birds performed better than vaccinated
unmedicated and medicated unvaccinated groups. The final mean weights of vaccinated medicated birds were significantly
higher (P < 0.05), and a better vaccine protection index, using both vaccinating methods, was achieved. Results indicated that
concomitant use of ionophores and vaccines could be a useful adjunct to planned immunization in the control of coccidiosis.
# 2005 Elsevier B.V. All rights reserved.

Keywords: Coccidiosis; Chicken; Ionophore-tolerant; Eimeria spp.; Vaccinated medicated; Vaccine protection index

1. Introduction Coccidiosis is an economically important parasitic

Coccidiosis is a disease of fowl caused by an
obligate microscopic protozoan parasite, which
belongs to the genus Eimeria (phylum Apicomplexa).
The disease is characterized by loss in weight
gain, increased feed conversion ratio, loss of skin
pigment and decreased egg production (Ruff, 1991).
* Corresponding author. Tel.: +86 20 85280241;
fax: +86 20 85282693.
E-mail address: gqli@scau.edu.cn (G.Q. Li).
disease of poultry resulting in losses exceeding between
US$ 1.5 and 2 billion annually to the poultry industry
worldwide (Stevens, 1998). In the UK alone, the total
cost of coccidiosis infections has been estimated to be at
least £42 millions per annum, of which 74% is due to
sub-clinical effects on weight gain and feed conversions
and 24% is the cost of prophylaxis and therapy of
commercial birds (Williams, 1998). As the world’s
poultry industry continues to grow, so do concerns about
coccidiosis which remains one of the most commonly
reported diseases of chickens. Coccidiosis is easily

0304-4017/$ – see front matter # 2005 Elsevier B.V. All rights reserved.
doi:10.1016/j.vetpar.2004.09.034
180 G.Q. Li et al. / Veterinary Parasitology 129 (2005) 179–186
transmitted by direct or indirect contact with droppings
of infected birds especially in intensively managed
poultry hence causing an inevitable choice of protecting
the birds against the disease.
Although coccidiosis can be controlled by pro-
phylactic chemotherapy, use of broad spectrum
anticoccidial agents and good management practices
and sanitary procedures, the emergency of rapid drug
resistance coccidia coupled with difficulties and high
costs of developing new drugs for effective treatment
and prevention has lead to a pressing need to search for
new approaches such as immunological, biotechno-
logical and genetic methods as a more promising
alternatives (Grag et al., 1999).
A variety of anticoccidial drugs have been
developed in the past with the ionophorous antibiotics
being the most extensively used groups, but resistance
has developed to almost all of them (Chapman, 1997),
and there are concerns about drug residues in poultry
products and strong desires of consumers to ban drugs
from animal feeds. The development of coccidial
resistance to drugs is probably the greatest single
factor causing demise of the effectiveness of drugs.
There is, therefore, a pressing need to move away
from chemotherapeutic control of coccidiosis towards
vaccination. Many anticoccidial vaccines, including
live virulent vaccines (e.g. CoccivacTM and Immu-
coxTM) and live attenuated vaccines (e.g. ParacoxTM
and LivacoxTM) are currently in use. The recent uptake
of attenuated coccidiosis vaccines worldwide has
given the poultry industry greater confidence that
vaccines may be used without any subsequent disease
problems that require attendant medication. The major
obstacle of these live vaccines (with either virulent or
attenuated parasites) is that they are usually adminis-
tered to chickens in the first week of age resulting in
low-level infection and depends on litter for the
recycling of vaccine parasites. During this period, use
of therapeutics or feed additives that interfere with
Eimeria development is not permitted, thus increasing
the risk of contracting the coccidiosis at an early (1–3
weeks) age and decreases as immunity has developed
(Vermeulen et al., 2001). Furthermore, Chapman
(1999) noted that acquisition of immunity in
medicated birds requires that the drug in question
permits some parasite development in the host and that
oocysts be present in the environment in sufficient
numbers to stimulate an immune response.
Recently, a new development has been made in the
use of live Eimeria strains, which are relatively
tolerant to ionophores (Danforth, 2000). The advan-
tage of these vaccines is that they allow the use of
ionophores during the first 3–4 weeks when the
immunity of the birds is not complete. This limits the
increase of infection pressure due to expanding field
strains during the development of immunity, hence
reducing the overall risk of contracting coccidiosis
(Vermeulen et al., 2001).
In this experiment, we investigate the immunolo-
gical responses of broiler chickens vaccinated with a
live attenuated ionophore-tolerant multivalent
Eimeria precocious line for protection against avian
coccidiosis.
2. Materials and methods
2.1. Chickens
Chinese Yellow disease-free chickens were pur-
chased from the South China Agricultural University
poultry farm, and used in the present experiment. They
were initially kept in a coccidian-free house and
transferred to wire cages for the experiments.
2.2. Coccidia vaccine
The vaccine is composed of a live attenuated
ionophore-tolerant stabilized suspension of multi-
valent sporulated oocysts of E. tenella, E. maxima and
E. acervulina (Li et al., 2004). The parasites were
isolated from chicken feces in a farm in Nanhai city,
Guangdong province, Southern China, by established
methods (Long, 1972). The three Eimeria species
isolated were then tested for drug sensitivity. Drug
resistant oocysts were sporulated and blended
aseptically into a suspension of 0.1% Carboxymethyl
cellulose sodium, which ensures the homogeneity of
oocysts and maintenance as suspension in drinking
water.
2.3. Challenge inocula
The material used to challenge the vaccinated
chickens were virulent strains of the same Eimeria
species in the vaccine. Each virulent strain had
 G.Q. Li et al. / Veterinary Parasitology 129 (2005) 179–186
previously been titrated using 55 susceptible Chinese
Yellow chickens in the pathogenecity test. The number
of sporulated oocysts needed in each inoculum, was
given by gavage, to produce a statistically significant
(P  0.05) reduction in weight gain during 1 week
after infection.
2.4. Vaccinations
Two vaccination trials were conducted using two
vaccination methods namely, by gavage and through
drinking water.
2.4.1. Vaccination by gavage
This test was carried out on 300 broiler chickens of
the Chinese strain (150 of each sex). Birds were
divided into five groups, with three replication of 20
birds (10 of each sex) kept on disinfected wire cages at
a stocking rate of 1 ft2 per bird. The first group was
vaccinated by gavage when the birds were 3 days of
age and a booster dose repeated a week later and fed
with rations containing 5% Monensin ionophore
vaccinated and medicated (VM). The second group
was vaccinated by similar method with a similar dose
as in group one but was not given feed containing any
therapeutic drug vaccinated unmedicated (VUM). The
third group was not vaccinated but was fed with
rations similar to those given to group 1 medicated
unvaccinated (MUV). The fourth and fifth groups
were maintained as positive and negative controls,
respectively (unvaccinated unmedicated challenged
control (UVUM-C), and unvaccinated unmedicated
non-challenge control (UVUM-NC)). All groups,
except group 5, were challenged 3 weeks later with
the virulent strains of the three Eimeria species
contained in the vaccinal material. Lesion scores were
determined 6 days after challenge according to the
method of Johnson and Reid (1970). In addition to
average weight gains, feed conversion ratio (FCR),
and oocysts per gram of feces (OPG) were also
determined and lesion score protection index (LSPI)
calculated according to the method of Danforth
(1998).
2.4.2. Vaccination by drinking water
A total of 360 Chinese Yellow chickens (180 of each
sex) were randomly divided, by equal weights, into five
groups of 12 birds each with three replications. Three
181
best vaccine batches (VB1, VB2 and VB3), determined
in the gavage vaccination trials were suspended in 0.1%
Carboxymethyl cellulose sodium solution for even
distribution of oocysts. The vaccine combinations of
three Eimeria species, E. tenella:E. maxima:E. acer-
vulina were 5000:2500:5000 in VB1, 2500:1250:2500
in VB2 and 1250:750:1250 in VB3 and the challenged
dose was 50,000:100,000:500,000. The vaccinal mate-
rial was then administered to groups 1–3 at 3 days of
age and a week later and fed with rations containing
100 mg/kg monensin anticoccidial drug. Groups 4 and 5
were maintained as unvaccinated unmedicated chal-
lenge and unchallenged control, groups, respectively.
All groups, except group 5, were later challenged 3
weeks later with a predetermined dose of the virulent
parent strain of the Eimeria species contained in the
vaccine. Lesion scores were again determined 6 days
post-challenge according to Johnson and Reid (1970)
and oocysts production per gram of feces determined to
calculate vaccine protection index (Rose and Mockett,
1983). Average live weight gains and FCR were also
determined at the end of the experiment.
2.5. Statistical analysis
The statistical analysis of weight gain, including
the various doses of vaccine combination used, was
based on Duncan’s multiple range test of analysis of
variance (ANOVA) using the SAS system (Version
8.1) statistical package. The factors included in the
ANOVA model are the mean weight gains and the
treatment comprising of VM, VUM, MUV, UVUM-
C and UVUM-NC in vaccination by gavage and
vaccine batch, VB1, VB2, VB3, UVUM-C and
UVUM-NC in vaccination by drinking water. The
group and animal effects are random, meaning that
they are a representative sample of all groups and
animals. By statistically modeling in this way, the
conclusions can be extended beyond the immediate
experiments to future batches and animals. The
factors sex and treatment are fixed effect factors. For
each Eimeria species, the mean weight gains per
batch of chickens in the five treatment groups (VM,
VUM, MUV, UVUM-C and UVUM-NC) were
compared. The statistical significance necessary to
demonstrate that the vaccinated birds were immune
is that the mean weight gain of the unvaccinated
unmedicated non-challenge groups need to be
182 G.Q. Li et al. / Veterinary Parasitology 129 (2005) 179–186

statistically significantly (P  0.05) lower than that
of the VM groups. Protective immunity of the
vaccinated birds was established when the mean
weight gain of the VM groups was statistically
significantly (P  0.05) greater than that of the
UVUM-C groups. In addition to the primary criterion
of weight gain, FCRs and lesion scores were
compared numerically when the same trend were
sought and used as a supportive secondary criterion
to weight gains.
3. Results
3.1. Vaccination by gavage
Table 1 show the summary of chickens vaccinated
by gavage with live attenuated ionophore-tolerant
multi-valent vaccine and/or medicated with antic-
occidial ionophore (monensin), and is illustrated in
Fig. 1. The vaccine protection index (VPI) was highest
for VM birds with indices of 86.93% and 88.07% for
male and female birds, respectively. There was no
significant differences between VUM and MUV
groups. The males of the VUM group had VPI
slightly higher (78%) than the females (76%).
However, the female birds of the MUV group had
higher VPI (72.61%) than the male birds (70%).
Considering the average oocysts produced per bird
after challenge, VM birds produced less oocysts 3
weeks after challenge. The VUM birds produced 3%
more oocysts than the MUV group, while the MUV
group produced about 1% more oocysts than the VUM
group. The plausible explanation is that VM birds
were well protected by deterring the establishment of
the parent oocysts, which were expelled much earlier.
As a result, few oocysts were able to establish
themselves without any adverse consequences on the
bird’s performances. Compared to the UVUM
challenge control group, VM produced 88% oocysts,
while VUM and MUV produced 85% and 86%,
respectively, less oocysts post-challenge at the end of
the first week. There was, however, a dramatic fall in
oocyst production in the subsequent 2 weeks that

Table 1
Summary of performance of Chinese Yellow broiler chickens immunized by gavage with live ionophore-tolerant multivalent Eimeria vaccine
and/or medicated with anticoccidial drugs
Group Treatment Sex FCR Lesion score Total oocyst production Post-challenge mean VPI (%)
(Av/(bird day)) weight gain (g)
UI MI C 7a 14b 21c 7d 14e 21f 28g Mean
1 VM M 1.98 0.2 0.2 0.4 36600 20000 6600 216.39 287.64 463.81 612.59 395.11A 86.93
F 2.26 0.3 0.3 0.3 33300 25800 8300 207.92 281.08 459.24 611.11 389.84A 88.07
2 VUM M 1.96 0.6 0.5 0.6 105100 47500 9900 216.25 258.06 441.19 497.62 353.28B 78.00
F 2.42 0.6 0.7 0.7 105000 51600 10900 212.64 254.31 419.76 459.28 336.50C 76.00
3 MUV M 1.92 0.8 0.7 0.8 142500 70000 16600 217.50 259.17 408.09 451.43 334.05C 70.00
F 2.48 0.8 0.8 0.8 134100 66600 27300 213.75 255.69 410.00 434.52 328.49C 72.61
4 UVUM-C M 4.66 3.0 3.0 3.0 268300 134100 68300 216.53 229.72 366.90 395.18 302.08D N/A
F 4.97 3.0 3.0 2.8 281800 185800 78300 215.14 224.31 388.81 398.81 306.77D N/A
5 UVUM-NC M 1.85 0.0 0.0 0.0 – – – 219.58 261.25 468.57 543.57 373.24B N/A
F 1.83 0.0 0.0 0.0 – – – 217.50 260.42 466.43 535.71 370.02B N/A
VM: vaccinated and medicated; VUM: vaccinated unmedicated; MUV: medicated unvaccinated; UVUM-C: unvaccinated unmedicated
challenged control; UVUM-NC: unvaccinated unmedicated non-challenged control; VPI: vaccine protection index; UI: upper intestine;
MI: middle intestine; C: caeca; N/A: not applicable. Means with the same letters (A–D) are not significantly different at a = 0.05 (DMRT).
a
Total oocyst produced (7 days) after challenge.
b
Oocysts produced (14 days) after challenge.
c
Oocysts produced (21 days) after challenge.
d
Mean weight gain (7 days post-challenged).
e
Mean weight gain (14 days post-challenged).
f
Mean weight gain (21days post-challenged).
g
Mean weight gain (28 days post-challenged).
 G.Q. Li et al. / Veterinary Parasitology 129 (2005) 179–186 183

followed. The FCR for VM groups was 1.98 for male

birds and 2.26 for females (n = 10). This was 7.23%
higher for VM male birds and 6.10% for VUM and
3.75% for MUV compared to that of the UVUM
female challenge control groups.

3.2. Vaccination by drinking water

Overall performance of chickens medicated and

Fig. 1. Post-challenge mean weights of chickens vaccinated by
gavage with ionophore-tolerant multi-valent Eimeria vaccine and/
or medicated with ionophore (monensin).
vaccinated with different vaccine batches (doses) by
drinking water is summarized in Table 2 and
illustrated in Fig. 2. After a week post-challenge,
there were no significant differences between unvac-
cinated unmedicated unchallenged control group and
birds vaccinated with vaccine batch 2. The perfor-
mances of birds vaccinated with vaccine batches 1
and 3 were significantly (P < 0.0001) less than the
unvaccinated unmedicated challenged control group,
but no apparent differences were observed between
sexes in each treatment group.

Table 2
Summary of performance of Chinese Yellow broilers chickens vaccinated by drinking water with live ionophore-tolerant multi-valent Eimeria
vaccine and/or medicated with anticoccidial drug (monensin)
Vaccine Treatment Sex FCR Lesion score Total oocyst production Post-challenged mean VPI (%)
batch (Av/(bird day)) weight gains (g)
UI MI C 7a 12b 17c 7d 14e 21f 28g Mean
1 VB1 M 3.96 1.2 2.5 1.5 44167 22500 27583 239.68 333.67 379.17 401.33 338.46B 62.00
F 4.06 1.5 1.0 1.6 39000 24167 8667 229.17 337.50 383.00 402.17 337.96B 60.00
2 VB2 M 1.96 0.9 1.5 0.8 19000 10333 3167 243.11 409.00 451.33 519.50 405.74A 80.30
F 2.22 0.6 0.7 1.0 18333 12167 4333 239.89 400.17 434.00 525.00 399.77A 77.00
3 VB3 M 3.92 0.8 0.7 1.7 31667 15167 10500 204.89 329.50 398.33 436.83 342.39B 57.00
F 4.48 0.8 0.8 1.6 35000 17250 9500 207.89 344.17 404.17 438.51 348.69B 59.00
– UVUM-C M 4.66 3.0 3.0 3.9 301000 164000 91500 241.17 368.00 388.50 400.75 349.61B N/A
F 4.97 3.0 3.0 3.9 304667 165000 91917 245.67 369.75 390.75 405.50 352.92B N/A
– UVUM-NC M 1.85 0.0 0.0 0.0 – – – 258.67 411.00 456.00 489.50 403.79A N/A
F 1.83 0.0 0.0 0.0 – – – 260.17 411.50 455.00 488.50 403.79A N/A
VB1: vaccine batch 1; VB2: vaccine batch 2; VB3: vaccine batch 3; UVUM-C: unvaccinated unmedicated challenged control; UVUM-NC:
unvaccinated unmedicated non-challenged control; VPI: vaccine protection index; UI: upper intestine; MI: middle intestine; C: caeca; N/A: not
applicable. Means with the same letters (A–D) are not significantly different at a = 0.05 (DMRT).
a
Total oocyst produced (7 days) after challenge.
b
Oocysts produced (12 days) after challenge.
c
Oocysts produced (17 days) after challenge.
d
Mean weight gain (7 days post-challenged).
e
Mean weight gain (14 days post-challenged).
f
Mean weight gain (21days post-challenged).
g
Mean weight gain (28 days post-challenged).
184 G.Q. Li et al. / Veterinary Parasitology 129 (2005) 179–186
Fig. 2. Post-challenge mean weights of chickens vaccinated by drinking water with ionophore-tolerant multi-valent Eimeria vaccine and/or
medicated with ionophore (monensin).
However, the unvaccinated unmedicated unchal-
lenged control group still outweighed 411.00 g and
411.50 g for males and females, respectively, com-
pared with birds vaccinated with vaccine batch 2 with
mean weights of 409 g and 400.17 g for males and
females, respectively. Comparing the three vaccine
batches, birds vaccinated with vaccine batch 2 had a
better performance with FCRs of 1.96 and 2.22, and a
vaccine protection index of 80.3% and 77%, for males
and females, respectively. This was followed by birds
vaccinated with vaccine batch 1 with FCRs of 3.96 and
4.06 and vaccine protection index of 63% and 60% for
males and females, respectively. In summary, vaccine
batch 2 with an Eimeria combination of 2500 oocysts
of E. tenella, 1250 oocysts of E. maxima and 2500
oocysts of E. acervulina provided the best protection
index (80.30%) for birds, hence considered superior to
the other vaccine combinations. It can therefore be
asserted, based on results obtained, that the best
vaccination dose of the vaccine for oral administration
by drinking water is 2500:1250:2500 oocysts of E.
tenella, E. maxima and E. acervulina, respectively.
4. Discussion
Previous laboratory and experimental studies
(Norton and Joyner, 1986; Shirley and Millard,
1986; Shirley, 1989) have demonstrated that a
multivalent vaccine containing precocious lines of
coccidia induces in chickens a strong immunity to
challenge with virulent homologous or heterologous
strains. Drug resistance has been documented against
almost all anticoccidial drugs used so far. Resistance
to polyether ionophorous anticoccidial has become a
major problem and there is growing evidence for a
significant shift in the effectiveness of these drugs. In
raising broiler chickens with a relative short life span
(40–45 days), the problem of drug residue has forced
farmers to comply with certain withdrawal periods for
the safety of consumers. In this research, a live
multivalent ionophore-tolerant anticoccidial vaccine
was developed and tested to evaluate the efficacy of
the vaccine during the growing period of broilers
(Chinese Yellow chickens strains) reared on wire
cages. The present study suggests that the concomitant
use of anticoccidial ionophores and live attenuated
ionophore-tolerant anticoccidial vaccines has the
potential to improve live weight gains and minimise
the spread of virulent field strain when the bird’s
immunity is not well developed, and subsequently
allow the gradual development of the bird’s local
immunity at a later date of the bird’s life span. This
dual approach strategy (chemotherapy and immuno-
prophylaxis) will certainly encourage the use of live
ionophore-tolerant anticoccidial in intensively broiler
production industries. It is on this basis that this
research was designed with the intent of addressing
drug resistant problems and providing adequate
protection against avian coccidiosis.
Comparison between the performances of broilers
either vaccinated or treated with anticoccidial drugs
 G.Q. Li et al. / Veterinary Parasitology 129 (2005) 179–186
reveal few consistent differences. These observations
tend to confirm the expectation that there is unlikely to
be any consistent improvement of bird’s weight gains
or FCRs by anticoccidial vaccines, unless drug
resistance has been a problem on a farm. However,
for some yet unexplained reasons, vaccinated birds
tend to have lower overall mortality than drug-treated
ones (Williams, 2002). Although live anticoccidial
vaccines may not offer any basic advantages in
performances when compared with drugs, there are
other benefits. A major advantage of vaccines is that
they cannot induce coccidial drug resistance. Indeed,
if resistant coccidia already exist on a farm, their
adverse effects may be ameliorated by the use of either
non-attenuated or attenuated vaccines. With respect to
studies on the performance of live anticoccidial
vaccines in broilers, there is no reason to expect that
a vaccine will perform better, under identical
conditions, than any non-ionophore anticoccidial
drug, unless the resident coccidia are drug resistant
(Williams, 2002). On the other hand, the performance
of vaccinated birds should be no worse than drug-
treated birds if the anticoccidial drug has no additional
therapeutic properties. However, in comparison with
ionophorous anticoccidial drugs, it is possible that
vaccinated birds might have poorer feed conversion
ratios (FCRs). This is possible because, even in
disease free chickens, ionophores used at recom-
mended concentration tend to improve FCRs (Wheel-
house et al., 1985).
The performance of birds vaccinated by gavage or
through drinking water and given monensin antic-
occidial drug and/or medicated unvaccinated or
vaccinated unmedicated were compared using the
following primary criteria: protection from clinical
coccidiosis, weekly bird weights, mean bird weights
after challenge, percentages of bird losses (culled or
found dead), total number of oocysts in feces, lesion
score protection index and feed conversion ratio.
Many scientists involved in coccidiosis research
have often questioned the criteria that is best to assess
chickens performance and appropriately evaluate drug
resistance. All criteria in this experiment have proved
to be of value under certain circumstances. Weight
gains measured the effects of coccidiosis upon growth
of the birds, lesion score estimated the pathology
caused by infection, and FCR measures the amount of
feed intake and conversion into animal tissues. Weight
185
gain has proven to be the most useful criterion for
evaluating the efficacy of anticoccidial drugs during
the acute phase of infection, and these may be
compared directly with unmedicated infected and
unmedicated uninfected controls. In this research
work, weight gains were similarly compared with
unvaccinated unmedicated challenged and unchal-
lenged controls. Lesion scores were also used but this
procedure is inherently subjective since it requires
different regions of the intestine, depending on the
Eimeria species involved, to ascertain pathology of the
parasite. It should also be remembered that, unlike
other criteria, lesion scores do not increase linearly
with number of oocysts inoculated (Chapman, 1994).
In this experiment, lesion scores were very much
erratic especially for species such as E. maxima and E.
acervulina. Lesion scores were, therefore, used as a
secondary criterion and as supportive evidence to
weight gains. A criticism of the use of lesion scores for
assessing coccidial infections is that under some
circumstances lesion scores do not correlate with
weight gain and those lesions may be present even
though weight gain is not depressed (Long and
Johnson, 1988). An interesting aspect of this broiler
vaccine is the ‘‘reaction’’ (the presence of coccidial
lesions and slowing of growth rate) that occurred with
some vaccine doses during the 2 weeks following
vaccination. However, the birds later exhibited a
compensatory weight gain that brought them up to
almost the same weight as unvaccinated unmedicated
unchallenged control groups by 5–6 weeks of age.
Moreover, because the birds may have acquired some
immunity, the reaction to challenge with homologous
virulent strains was minimal compared to unvacci-
nated unmedicated challenged controls. The live
anticoccidial vaccine developed and tested in this
experiment proved to be efficacious and provided
satisfactory protection for the chickens. The vaccine,
if administered by gavage, requires a less dosage of
attenuated oocysts but a slightly higher dosage is
needed when administered through drinking water.
Acknowledgements
This research was partially financed by the Council
of State Education Committee of the People’s
Republic of China and by Natural Science Fund of
186 G.Q. Li et al. / Veterinary Parasitology 129 (2005) 179–186

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