Dept. of Medicine,
Fac. Vet. Med. , Zagazig University, Bertha Branch.
IMMUNE RESPONSE TO FOOT AND MOUTH DISEASE VACCINE
IN GUINEA PIGS AND CALVES EXPERIMENTALLY
INFESTED WITH TRYPANOSOMA EVANSI
(With 2 Tables)
By
A.A. EL-SAWALRY' N.H. ABD EL-SAMEA; F.K. HANODA;
NAi;ED HETWALLY and SOHEIR BANOUB
(Received at 25/5/1994)
SUMMARY
The immune response to Foot and Mouth disease vaccine was
investigated using serum neutralization test on 25 guinea
pigs,
and
12
calf
sera
experimentally
infected
with
Trypanosoma evansi. In guinea pigs, there was a significant
decrease in mean FMD neutralizing antibody titers in those
animals
inoculated
with
T.evansi
and
FMD
vaccine
simultaneously as well as those inoculated with T.evansi one
week before vaccination. Furthermore a highly significant
decrease was observed for guinea pigs inoculated with
trypanosomes two weeks before vaccination, compared to
uninfected control guinea pigs which received only the
vaccine. Regarding the immune response for calves, there was a
significant and a highly significant decrease in mean serum
neutralizing
antibody
titers
for
animals
injected
simultaneously
and
two
weeks
before
FMD
vaccination
respectively. It can be concluded that T.evansi infection
induced an immunosuppressive effect in animals vaccinated with
FMD vaccine. So it is recommended that animals must be
examined
and
be
free
from
T.evansi
infection
before
vaccination with FMD vaccine.
INTRODUCTION
Trypanosomiasis caused by Trypanosoma evansi is considered as one
of the most important disease affecting domestic animals camels, cattle,
buffaloes, sheep, goats, pigs and mules in Africa and Asia, usually show
a chronic disease, but in horses, deer and dogs the disease is more
acute and rapidly fatal (LUN et al., 1992). The pathogenicity of certain
infectious diseases
6th Scl. Cong., 20-22 Nov. 199C, Fac. Vet. Ned., Assiut, Egypt.
304
c
may arise as a result of a suppression in the host's immune
system by some unrelated disease entity (KREIER and BAKER,
1992).
Infection
with
many
parasitic
protozoa
causes
immunosuppresion. It has been demonstrated that Babesia
microti infection can suppress the humoral response of mice to
sheep red blood cells (RUEBUSH and HANSON, 1980). Also in
mice Babesia hylomysa has been shown to depress the immune
response to Trichuries muris (PHILIPS and WAKELIN, 1976).
There
have
been
few
reports
of
piroplasm
induced
immunosuppression in domestic animals. CALLOW and STEWART
(1978) demonstrated that Babesia bovis infestation in cattle,
causes immunosuppresion against its tick vector Boophiiis
microplus, and WAGNER et al., (1975) demonstrated suppression
of the rinderpest neutralizing antibody response in cattle
severely ill with East Coast Fever (Theileria
parava
infection). Furthermore SHARPE and LANGLEY (1983), showed
that cattle infested with Theileria annulata response weakly
to FMD vaccine compared to uninfected control cattle. It has
been shown that goats infested with Trypanosoma congolense,
had a significant, but not complete suppression of antibody
response to Brucella melitensis which was most marked with the
onset of parasitaemia (GRIFFIN et al., 1980).
The ability of African salivarlan trypanosomes, to induce
immunosuppression in their hosts is well documented following
studies on laboratory animals (VICKERMAN and BARRY, 1982 and
ROELANTS and PINDER, 1984). Vaccination against either
Louping ill virus or lymphocytic choriomeningitis virus, of
groups of trypanosome infested mice, showed that T. cell
responses are only diminished, and major immunosuppressive
effect
of
trypanosomes
is
brought
about
by
B-cell
unresponsiveness (REID et al., 1979). HOLMES et al. (1974)
recognized that the secondary immune response to polyvalent
clostridial vaccine, was lower in cattle infested with
T.congolense than uninfected control animals. Furthermore
RURANGIRWA et al. (1978) demonstrated that cattle in Kenya
artificially infested with T.vivax and T.congolense, were
suppressed in their response to the antigenic stimulus of
Mycopiasma mycoids, the causative agent of contagious bovine
pleuro-pneumonia (CBPP), these workers suggested that the
involution of the thymus in trypanosome infestations may be a
contributory factor.
This
work
was
planned
to
study
the influence
of
experimental infestation with Trypanosoma evansi (for guinea
pigs and calves) on the immune response following vaccination
with FMD vaccine.
6th Sc!. Cong., 20-22 Nov. 1994, Fac. Vet. Ned., Assiut, Egypt.
305
MATERIAL and METHODS
Animals
1- Guinea pigs:
Twenty five,
apparently
healthy
adult
albino
male
guinea
pigs
of
approximately 500 grams were used. The animals were free from blood parasites
by blood examination before use, and divided into five groups each of S
animals.
6
Group 1. was inoculated intraperitoneally with lx10 virulent field strain
of T.evansi according to BHATTACHORYYA and SINHA (1974).
Group 2. was inoculated simultaneously with 1x106 virulent field strain of
T.evansi and with 1 ml of foot and mouth disease vaccine subcutaneously.
Grup 3. was inoculated with the same number of trypanosomes one week before
vaccination with (RID) vaccine as in group 2.
Group 4. was inoculated with the same number of trypanosomes two weeks
before vaccination with (FMD)vaccine. Group 5. was vaccinated with (FMD)
vaccine only.
2. Calves:
Twelve mixed breed calves of one year old, were obtained from the farm of
Faculty of Veterinary Medicine, Suez Canal University. These calves were
proved to be free from external, and internal parasites including blood
parasites by parasitological examination, Furthermore calves proved to be
free from neutralizing antibodies againest FMD virus. These animals were
divided into three groups each of four calves as follows:
Group 1. was inoculated simultaneously, with 5x107 virulent field strain of
T.evansi subcutaneously according to SING)! and GAUR (1983) and 2cc of FMD
vaccine subcutaneously.
Group 2. was inoculated with the same number of T.evansi two weeks before
vaccination with FMD vaccine as in group 1. Group 3. was vaccinated with
FMD vaccine only.
Foot and Mouth disease vaccine:
Tissue culture formaline inactivated vaccine, against FMD virus type 01
was prepared at Veterinary Serum and Vaccine Research Institute, Abbassia,
Cairo, Egypt as described by NOUSSA et al. (1974). The dose was 1, 2 ml for
each guinea pigs and calf respectively to be given subcutaneously.
Trypanosomes:
Trypanosma evansi was isolated from infested camel at Balbies, and were
kept in mice until animal inoculation, and the parasitaemia were performed using
haemocytometer.
6th Scf. Cong., 20-22 Nov. 1994, Fac. Vet. Mcd., Assiut, Egypt.
306
Serum samples:
Blood samples were collected from medial canthes of the eyes of
guinea pigs, weekly for 4 weeks post vaccination, and from the
jugular vein of calves before vaccination with FMD vaccine and weekly
post vacciation for four weeks.
Serum neutralization test (SNT):
Serum neutralization test was performed in a BWC cell cultures
according to the technique described by HABEL (1969).
RRESULTS
Table I. Mean serum neutralizing FMD antibody titers for
guinea pigs expressed in Log10 TCID56'ml
Times post Group I Group 2
vaccination ?dean±SD. .\tean±SD.
Group 3
bfean±SD
Group 4
\iean±SD
Group 5
(control)
',lean±SD.
First week 00•
Second Week 00
Third Week 00
Fourth week 00
0604±0 119+t 0.656±0100* 0 53±0 1050 0 769±0 155
1058±0.157
1.03130 150 1.125±0 106 1 363±0 192
1.200±0.138 1.175 ±112 L150±0.087 1.525±0 126
1375±0 152
1 299±0 201
1
.236}0 178 I Ja 5±0 218
Log10 TCID50 mean Logarithm 10 tissue culture infective dose 50
• Negative serum neutralizing antibody titer
*0. test) 0 01 <P<0.05
•(t. test) P<0 01
The results for guinea pig showed a significant decrease in
neutralizing antibody titers in Group 2 and 3 (both Groups inoculated
with T.evansi simultaneously with vaccine and one week before
vaccination respectively), compared to Group 5, and also a highly
significant decrease in Group 4 which received FMD vaccine two weeks
after experimental inoculation of T.evarlsi compared to control Group
5. Also in the control Group the highest titre of FMD neutralizing
antibody titers was shown during the 4 weeks post-vaccination.
Table 2. Mean serum neutralizing antibody titers for calf
sera expressed in Logt TCIDSOimt
Time post
vaccination
Group I
Sfean±SD.
Group 2
First week
1.150±0.091 *
%lean±SD.
1
2750 065
Second Week
Third Week
1 250±0 127
1 350±0 120
0 887±0 125
•
1 725±0 119
1.250±0 100
Fourth week
1425±0.065
1 350±0 108
1.650±0.130
%Iean±SD
Group 3
(control)
1.337±0085
1 SOO±0 129
*(t test) 0.01
<P<0 05 •(t. test)
P<0.01
i
6th Set. Cong., 20-22 Mov. 1994, Fac. Vet. Med., Assiut, Egypt.
307
The results demonstrated that there was a significant decrease in
mean neutralizing antibody titers in group 1 compared with that shown
group 3. Furthermore there was a highly significant decrease in Group
2.
DISCUSSION
Trypanosomiasis caused by T.evansi infestation is a common and
economically important infection among domestic animals in Africa and
Asia, since the disease Is usually chronic in nature (LUN et al., 1992).
The protective immunity in vaccinated cattle is generally considered to
be closely related to the serum neutralizing antibodies present at the
time of exposure to infection and there is no published evidence that
cell mediated immunity plays a role in protection induced by FMD
vaccination (PAT, 1989).
1)
It is evident from the results obtained in the present study (Table
that; all guinea pigs vaccinated with FMD vaccine induced
neutralizing antibody titers in their sera, as measured by serum
neutralization test. Guinea pigs received trypanosomes simultaneously
with FMD vaccine (Group 2) as well as those animals injected with
T.evansi one week before vaccination (Group 3), showed a significant
decrease in FMD neutralizing antibody titers in their sera compared
with those animals of group 5 which received only FMD vaccine.
Furthermore a highly significant decrease was observed in Group 4
inoculated with trypanosomes two weeks before vaccination.
The nature and extent of immunosuppression in laboratory animals
infested with trypanosomes has been investigated and reviewed by
MANSFIELD (1978) and URQUHART (1980) and a number of theories have been
proofed to explain its occurrence such as: B cell mitogen preempting
response to antigens; action of suppressor T-cells or macrophages; loss
of suppressor cell function with plasma cell hyperplasia, increased
catabolism of immunoglobulin; production of free fatty acids and
effects of immune-complexes on phagocytic function.
Our results for calves (Table 2) indicated that those calves
receiving trypanosomes simultaneously with FMD vaccine (Group 1)
revealed a significnt decrease in FMD neutralizing antibody titers in
their sera compared with those control animals which received only FMD
vaccine (Group 3). Those animals receiving T.evansi two weeks before
vaccination (group 2) showed a highly significant decrease in mean
neutralizing antibody titers. These results supported the results
observed in guinea pigs.
These observations agreed with one of HOLMES et al. (1974) whho
recognized that the secondary immune response to polyvalent clostridial
vaccine was lower in cattle infested with a closely related organism
T.congolense than in uninfested
6th Sc f. Cong., 20-22 Nov. 2994, Fat. Vet. Med., Ass fut, Egypt.
308
control animals. Furthermore our results run parallel with these observed by
RURANGIRWA et al. (1978) and GRIFFIN et al. (1980).
Concerning the protective serum neutralizing antibody titers BAA'GELSDORFF
(1989) reported that the serum neutralizing antibody titer protective level of
FMD vaccine ranged between 0.9 and 1.5. It is evident from Table 2 that serum
neutralizing antibody titers in Group 2 was non protective by the first week
post-vaccination. Furthermore LUCAM et al. (1969) demonstrated that serum
neutralizing antibody titer < 1.5 Logio was protective for only 75% of cattle
against severe FMJ virus infection. From our results in Table 2 animals in group
I and 2 showed serum neutralizing antibody titer < 1.5 Logio in their sera, so
25% of these animals were liable to FMD virus infection.
In conclusion Trypanosoma evansi induced on immunosuppressive effect for
animals vaccinated with FMD vaccine; so it is recommended to treat the animals
with trypanocidal drugs before vaccination to obtain protective antibody titers
post-FMD vaccination.
REFERENCES
Bangelsdorff, H.J. (1989): Potency test of Foot and Mouth Disease vaccine.
Correlation between response to challenge and corresponding neutralizing
antibody titers of vaccinated cattle. Berl. Munrch. Tierarztl Wschr, 102:
193-198.
Bhattachoryya, U.K and Sinha, P.K. (1974): Behavior of experimental infection of
Trypanosoma evansi in guina pigs. Indian Journal Animal Health 13(1): 4751.
Callow, L.L. and Stewart, N.P. (1978): Immunosuppression by Babesia bovis
against its vector Boophilus microplus. Nature, London, 27k: 818-819.
Griffin, L.; Waghela, S. and Allonby, E.W. (1980)v The immunosuppressive effects
of experimental T.congolense infections in goats. Veterinary Parasitology,
7: 11-18.
Babel, K.
(1969): Virus Neutrlization Test.
Fundamental
techniques in Virology. Academic Press, NY.
Holmes, P.H.; Nammo, E. and Thomason, A. (1974): Immunosuppression in bovine
Trypanosomiasis. Veterinary Record, 95: 86-92.
Kreier, J.P. and Baker, J.R. (1992): Parasitic Protozoa. Academic Press, NY.
Lucam, F.; Fedida, N. and Dannacher, C. (1969): Measure de L. immunite antiaphteuse post-vaccination du beof par epreuve sur le cobbaye. Rev. Med.
Vet., 115: 225-245.
Lun, Z.R.; Al l Ingham, R.; Brun, R. and Lanham, S.H. (1992): The isoenzyme
characteristics of Trypanosoma evansi and Trypanosoma equiperdum isolated
from domestic stocks in
6th Sci. Cong., 20-22 Nov. 1994, Fac. Vet. Med., Assiut, Egypt.
----------------------------------------------------------------309
China. Annals of Tropical Medicine and Parasitology, Vol. 86, No.
4: 333-340.
Mansfield,
J.M.
(1978):
Immunobiology
of
African
trypanosomiasis. Cellular immunology, 39: 204-211.
Moussa, A.A.M.; Stouraitis, P.; Ibrahim, H.H.; Daoud, A. and Hussein, K.
(1974): Foot and Mouth Disease (FN.D) vaccine production in baby
hamster kidney cells (BHK2I clonel3) in suspension in Egypt. Bull.
Off. Int. Epiz., 81: 1043-1054.
Pat, T.W.F. (1989): Control of Foot and Mouth Disease in cattle by
vaccination. Vet. Bulletin, 27(2): 1-9.
Philips, R.S. and Wakelin, D. (1976): Trichuris muris effect of concurrent
infections with rodent piroplasms on immune expulsion from mice.
Experimental Parasitology, 39: 95-100
Reid, H.W.; Hdmes, P.H. and Skinner, H.H. (1979): Immunosuppression in
experimental trypanosomiasis effects of Trypanosoma brucei on
immunization
against
Louping-ill
virus
and
lymphocytic
choriomeningitis virus. Journal of Comparative Pathology, 89: 581586.
Roelants,
C.E.
and
Finder,
M.
(1984):
Immunobiology
of
African
Trypanosomiasis. Contemp. Top. Immunobiol., 12: 225-274.
Ruebush, M.J. and Hanson, W.L. (1980): Thymus dependence of resistance to
infection with Babesia microti of human origin in mice. American
Journal of Tropical Medicine and Hygiene, 29: 507-515.
Rurangirwa, F.R.; Tabel, H.; Losos, G.; Masiga, W.N. and Mwambu, P. (1978):
Immunosuppressive effect of Trypanosoma congolense and Trypanosoma
vivax on the secondary immune response of cattle to Mycoplasma
mycoides sub sp. mycoides. Research in Veterinary Science., 25:
395-403.
Sharpe, R.T. and Langley, A.M. (1983): The effect of Theileria annulata
infecion on the immune response of cattle to Foot and Mouth
disease. British Veterinary Journal., 193,5: 378-385.
Singh, D. and Gaur, S.N.S. (2983): Clinical and blood cellular changes
associated with Trypanosoma evansi infection in buffalo calves.
Indian Journal of Animal Science, 53,5: 498-502.
Urquhart, G.M. (1980): The effect of trypanosomiasis on the immunological
apparatus. Transactions of the Royal Society of Tropical Medicine
and Hygiene, 67: 528-535.
Vickerman, K. and Barry, J.D. (1982): African trypanosomiasis. In:
Immunology of parasitic Infections. (S. Cohen and J.S. Warren,
eds.), pp. 204-260. Blackwell Scientific Publications, Oxford.
Wagner, G.G.; Jessett, D.M.;
Brown, C.G.D. and Radley, D.E.
(1975): Diminished antibody response to rinderpest vaccination in
cattle undergoing experimental East Coast fever. Research in
Veterinary Science, 19: 209-211.
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