Research Article
Open Access
T Lymphocytes Response to Caecal Coccidiosis in Broilers Infected with Exo and Endogenous Stages of Eimeria Tenella
Kaze Paul Davou1*, Idris Lawal2, Ajanusi Joseph2, Saidu Lawal3, Gloria Karaye1, Maryam Patrobas1, Biallah Markus1, Francis Sa Ayinzat4, Dung Christopher5, Kuduk Karaye6
1Department of Veterinary Parasitology and Entomology, Faculty of Veterinary Medicine, University of Jos, Nigeria
2Department of Veterinary Parasitology and Entomology, Faculty of Veterinary Medicine, Ahmadu Bello University, Zaria, Nigeria
3Department of Veterinary Medicine, Faculty of Veterinary Medicine, Ahmadu Bello University, Zaria, Nigeria
4Department of Theriogenology and Production, Faculty of Veterinary Medicine, University of Jos, Nigeria
5Department of Veterinary Biochemistry and Physiology, Faculty of Veterinary Medicine, University of Jos, Nigeria
6Central Diagnostic Laboratory, National Veterinary Research Institute, Vom, Nigeria
2Department of Veterinary Parasitology and Entomology, Faculty of Veterinary Medicine, Ahmadu Bello University, Zaria, Nigeria
3Department of Veterinary Medicine, Faculty of Veterinary Medicine, Ahmadu Bello University, Zaria, Nigeria
4Department of Theriogenology and Production, Faculty of Veterinary Medicine, University of Jos, Nigeria
5Department of Veterinary Biochemistry and Physiology, Faculty of Veterinary Medicine, University of Jos, Nigeria
6Central Diagnostic Laboratory, National Veterinary Research Institute, Vom, Nigeria
*Corresponding author: Department of Veterinary Parasitology and Entomology, Faculty of Veterinary Medicine, University of Jos, Nigeria, E-mail:
@
Received: November 09, 2017; Accepted: January 19, 2018; Published: January 22, 2018
Citation: Davo KP, Lawal I, Joseph A, et al. (2018) T Lymphocytes Response to Caecal Coccidiosis in Broilers Infected with Exo and Endogenous Stages of Eimeria Tenella. SOJ Immunol 6(1): 1-6. DOI: http://dx.doi.org/10.15226/2372-0948/6/1/00166
Summary
The present study reveals the proliferation of cytokines in treated and non- treated broilers consisting of IFN- γ, IL-1, IL-2, IL-4, IL-6, TNF and TGF. The CD4 count in the treated and non- treated broilers orally administered with various developmental stages of the parasite reached a peak on day 10 at primary; secondary and day 24 at tertiary infections. There was significant difference in the CD4 cell count between groups of the infected broiler chickens (p < 0.05). The current study observed a relationship between the different developmental stages of the parasite and lymphocytes response. Broiler chickens infected with sporulated oocyst (sporozoites) and merozoites treated and non-treated gave high CD4 T- lymphocyte numbers than the other groups throughout the experimental periods.
Introduction
Eimeria tenella is the most prevalent and pathogenic coccidian resulting in morbidity and mortality, causing serious economic losses to the poultry industry worldwide. Immunocompetence is a key factor in evolutionary process given that parasitism and disease are strong forces promoting genetic variability, May and Anderson, et al [1]. A decrease in the immune response when experimentally exposed to parasites has been interpreted as a depletion of resources devoted to immune stimulation [2]. However, an increase in the immune response after parasite infection is considered to be due to the stimulation of host immunological activity [3,4]. In birds, the most abundant and studied T- lymphocyte subsets are CD4, CD5 and CD8 as they played a role in active defense against infections [5,6]. CD4 subset is implicated in the secretion of active substances such as cytokines, interferon and interleukin-6 and commonly used to evaluate the health status of infected birds with infections [7]. This study aim is to determine the T-lymphocyte response in broilers infected with exo and endogenous stages of Eimeria tenella.
Materials and Methods
Materials and Methods
The experimental settings was at the PETCA building, Anguldi, 5 kilometers from the National Veterinary Research Institute, Vom, Jos Plateau State, Nigeria, where the laboratory work was carried out. The Jos Plateau lies on the pre-cambian from the cambian to jurasic northern Nigeria crystalline complex in central Nigeria. Its average elevation is about 1,250 m above mean sea level. The state is bounded on the north and west by Kaduna plains (on the average of 600 m above mean sea level) and on the south by Benue plains (on the average of 700 m above mean sea level), [8]. Geographically, the Jos Plateau is located between latitude 08°24’N and longitude 008°32’ and 010°38’ east. The land surface of Jos Plateau consists of plains, hills, depressions and todes of various forms, shapes and sizes. It is a major tourist centre in Nigeria with agriculture as the main occupation of the people. The high altitude confers on the Plateau lower temperature than those encountered elsewhere in Nigeria except the Obudu and Mambilla Plateau. The dry season is determined by the north easterly tropical continental air masses known as harmattan (from October – April) and the wet season is the most tropical maritime air masses from May – September. The average annual rainfall is about 1,100 mm and is evenly distributed. Another element of climate is temperature December and January experience temperatures of below 150C. During February and March, the temperature rises again about 250C. Most of the human activities are mining and agriculture involving rearing of chickens in both the rural and urban areas
for subsistency and income [8].
Experimental Birds
Four hundred (400) day-old broilers (marshal breed) were
purchased from ECWA farms, Jos, brooded and used for the study.
The birds were randomly distributed into six different groups of
40 each, in a clean wire cage (n = 40). At two weeks old, each
group was again subdivided into two, treated and non treated,
of twenty broilers (n = 20) each. The birds were kept in a clean
building, and the legs banded or labelled under strict biosecurity
measures. Feed (Broiler starter, Grand cereals and oil mills, PLC,
Zawan, Jos-Plateau, Nigeria) and water were provided adlibitum
The birds were vaccinated with Newcastle disease vaccine (La-
Sota) at day 21 and Gomboro disease vaccines at days 14 and 28.
Experimental infection of broilers with infective
materials and monitoring
The experimental birds, except the control were orally
given primary and secondary challenge infections with the
various developmental stages of Eimeria tenella, respectively at
week 2 and 3 ½ while at week 5 of age, all birds were infected
the sporulated oocyst of the parasite (Table 1). Each group was
subdivided into Treated (n = 20) and Non- Treated (n = 20). In
each infected group, birds in one of the subdivisions were treated
with amprolium 250 WSPR Holland was administered in drinking
water at a concentration of 250 mg/1 (0.025%) for a period of
5 days as prescribed by the Manufacturer at the appearance of
visible clinical signs.
To obtain serum, blood samples were collected from the experimental birds using the method described by Talebi and Mulcahy [9]. Briefly, 1ml of blood sample was obtained from the wing vein of each bird using 20 gauge needle (Becton Dickson co., Plymouth, UK) into a 2ml vacutainer. Samples were obtained on days 2, 4, 6, 8, and 10 after primary and secondary infections, and on days 5, 7, 11, 14, 17, 20 and 24 after tertiary infection (Rose and Hasketh, et al). The blood which had been allowed to clot for 1 hour at room temperature, was left over night at 40C and then centrifuge at 800g for 5 minutes. The serum samples were thereafter heated at 560C for 30 minutes to inactivate the compliment before storage at -200C. All sera were analyzed with the developed ELISA Triplicate
To obtain serum, blood samples were collected from the experimental birds using the method described by Talebi and Mulcahy [9]. Briefly, 1ml of blood sample was obtained from the wing vein of each bird using 20 gauge needle (Becton Dickson co., Plymouth, UK) into a 2ml vacutainer. Samples were obtained on days 2, 4, 6, 8, and 10 after primary and secondary infections, and on days 5, 7, 11, 14, 17, 20 and 24 after tertiary infection (Rose and Hasketh, et al). The blood which had been allowed to clot for 1 hour at room temperature, was left over night at 40C and then centrifuge at 800g for 5 minutes. The serum samples were thereafter heated at 560C for 30 minutes to inactivate the compliment before storage at -200C. All sera were analyzed with the developed ELISA Triplicate
Table 1: Experimental infection of broilers with developmental stages of Eimeria Tenella
Group |
Treatment and No. of birds |
Infection type/ Age of bird |
||
I° /wk 2 |
2° /wk 3 ½ |
3° /wk 2 challenge with virulent E. tenella |
||
I |
T(n=20) |
105 USO |
105 USO |
105 SO |
NT(n=20) |
105 USO |
105 USO |
105 SO |
|
II |
T(n=20) |
105 SO |
105 SO |
105 SO |
NT(n=20) |
105 SO |
105 SO |
105 SO |
|
III |
T(n=20) |
105 SCZ |
105 SCZ |
105 SO |
NT(n=20) |
105 SCZ |
105 SCZ |
105 SO |
|
Iv |
T(n=20) |
105 MRZ |
105 MRZ |
105 SO |
NT(n=20) |
105 MRZ |
105 MRZ |
105 SO |
|
V |
T(n=20) |
105 GMT |
105 GMT |
105 SO |
NT(n=20) |
105 GMT |
105 GMT |
105 SO |
|
VI |
|
0 |
0 |
0 |
USO: Unsporulated oocyst; 1°: primary infection; SO: Sporulated oocyst; 2°: Secondary infection; SCZ: Schizoites; 3°: Tertiary infection;
WK:Week; T: Treated; NT: Non treated.
Determination of Immunity Conferred on Birds By
Eimeria tenella Developmental Stages
This was done by measurement of immune bodies using
Lymphoproliferation assay or Non-radioactive assay and Flow
cytometric analysis
Lymphocyte Proliferation Studies
The lymphocyte proliferation assay is widely used to evaluate
cell-mediated immunity in normal and disease states in chickens
[10,11]. The spleen collected from each of the sacrified broilers
(four per group) at the end of each infection were crushed by
pressing on fine mesh Petri dishes containing PBS and glass
beads. The suspension was then passed through nylon cell
strainer (70 μm; Becton, Dickson, Lincoln Park, NJ). The filterate
was centrifuged at 250 g for 10 minutes at 4°C and the sediment
containing the spleen cells was collected for the study. One
hundred microlitre of splenic cell suspension containing 5 × 105
cells was placed in each of 96-well sterile culture plate (Corning,
NY) containing 100 μl of complete Rose Park Memorial Institute
(RPMI) media containing various concentrations (0.1, 1.0, 10
or 20 μg/ml).of concanavalin A (Con-A, Sigma, MO). The plates
were incubated for 48 hours at 37°C in a 5% CO2, 95% humidity
incubator [12].
Colorimetric Analysis
After 48 hours of culture of the splenic cells of the orally
infected broiler chickens, Resazurin or Alamar Blue TM
(Accumed International, Westlake, OH, from Biosource/Tago
Immunochemicals, Camars, CA) was added at 20 μl/well, and
absorbance value were read at wavelengths of 570 nm (reduced
state) and using an optical density Colorimeter Plate Reader
(Molecular Devices, Menlo Park, CA) 24 h after the addition of
Alamar Blue. Purple colour was observed on the proliferated
lymphocytes [13].
Flow Cytometric Analysis
Whole blood (20 μl) from each group of treated and non
treated broilers orally infected with the parasite stages was added
into a test tube, cells were stained with fluorescein isothiocyanate
(FITC)-conjugated anti-mouse CD90.2 (Thy1.2), isotype anti-rat
IgG2a, κ control antibodies; and kept in the dark for 15 minutes at
room temperature. These stained cells were analyzed using CD4/
CD4% PARTEC CYFLOW-Cyflow counter 2010, USA [13].
Data Analyses
The data were subjected to statistical analysis using Analysis
of variance (ANOVA) by SPSS version 20. Data was expressed as
(mean ± standard error of the mean (mean ± S.E.M). Values of p <
0.05 was considered significant.
Results
Lymphoproliferation studies in the experimental
broiler chickens
Lymphoproliferation assay showed the proliferation of
protective cytokines in the spleen of broilers infected with the
various developmental stages of the parasite. Groups I (infected
with unsporulated oocyst) and III (infected with schizonts) did
elicit lymphocytes proliferation (IFN- γ, IL-1, IL-2, IL-4, IL-6
and TNF) at primary and secondary infections respectively.
Groups II and IV revealed the proliferation of Interferon (IFN- γ),
Interleukins ( IL-1, IL-2, IL-4, IL-6), Turmor necrotic factor (TNF),
Transforming growth factor (TGF), respectively at primarysecondary-
tertiary infections, while Group V (infected with
gametocytes) showed IL-1, IL-2 and IL-4 at primary and secondary
infections. However, groups I, III and V showed proliferation of
IFN- γ, IL-1, IL-2, IL-4, IL-6, TNF and TGF respectively at tertiary
infection. The control showed no lymphocytes proliferation.
In the study, the number of CD4 count increased post infection in treated and non-treated broilers orally administered with various developmental stages of the parasite, reaching a peak at day 10 ((groups I – 198.0 x 103 μl, 165.3 x 103 μl; 200.0 x 103 μl, 156 x 103 μl and 196.7 x 103 μl, 173.3 x 103 μl; II – 199.0 x 103 μl, 186.0 x 103 μl; 197.0 x 103 μl, 192.7 x 103 μl and 200.0 x 103 μl, 194 x 103 μl; III – 198 x 103 μl, 153.3 x 103 μl; 200.0 x 103 μl, 160.0 x 103 μl and 188.7 x 103 μl, 166.7 x 103 μl; IV – 193.3 x 103 μl, 183 x 103 μl; 198.7 x 103 μl, 183.3 x 103 μl and 190 x 103 μl, 188.0 x 103 μl; V – 200.0 x 103 μl, 198.0 x 103 μl; 187.3 x 103 μl, 174 x 103 μl and 188.7 x 103 μl, 175.3 x 103 μl respectively) of primary and secondary infections and day 24 of tertiary infection (Tables 2.1, 2.2 and 2.3). There was significant difference in the CD4 cell count among different groups of the experimental broilers ( p < 0.05 ). CD4 levels were higher in the treated than the non treated broilers at primary infection (Figure 1.1). The levels of CD4 cells increases rapidly in the non treated birds at secondary infection, showing a non significant difference in the CD4 levels in all the groups treated and non treated (Figure 1.2). Groups II and IV of the non treated birds had higher CD4 levels than groups I, III and V at both secondary and tertiary infections (Figures 1.2 and 1.3). There was significant difference in CD4 subset between groups of the study birds (p < 0.05). The current study observed a relationship between the different developmental stages of the parasite and immune responses ( humoral and lymphocytes responses). Broilers infected with sporulated oocysts (sporozoites) and merzoites yielded high CD4 T-lymphocyte numbers than the other groups, throughout the experiment periods (Tables 2.1, 2.2 and 2.3).
In the study, the number of CD4 count increased post infection in treated and non-treated broilers orally administered with various developmental stages of the parasite, reaching a peak at day 10 ((groups I – 198.0 x 103 μl, 165.3 x 103 μl; 200.0 x 103 μl, 156 x 103 μl and 196.7 x 103 μl, 173.3 x 103 μl; II – 199.0 x 103 μl, 186.0 x 103 μl; 197.0 x 103 μl, 192.7 x 103 μl and 200.0 x 103 μl, 194 x 103 μl; III – 198 x 103 μl, 153.3 x 103 μl; 200.0 x 103 μl, 160.0 x 103 μl and 188.7 x 103 μl, 166.7 x 103 μl; IV – 193.3 x 103 μl, 183 x 103 μl; 198.7 x 103 μl, 183.3 x 103 μl and 190 x 103 μl, 188.0 x 103 μl; V – 200.0 x 103 μl, 198.0 x 103 μl; 187.3 x 103 μl, 174 x 103 μl and 188.7 x 103 μl, 175.3 x 103 μl respectively) of primary and secondary infections and day 24 of tertiary infection (Tables 2.1, 2.2 and 2.3). There was significant difference in the CD4 cell count among different groups of the experimental broilers ( p < 0.05 ). CD4 levels were higher in the treated than the non treated broilers at primary infection (Figure 1.1). The levels of CD4 cells increases rapidly in the non treated birds at secondary infection, showing a non significant difference in the CD4 levels in all the groups treated and non treated (Figure 1.2). Groups II and IV of the non treated birds had higher CD4 levels than groups I, III and V at both secondary and tertiary infections (Figures 1.2 and 1.3). There was significant difference in CD4 subset between groups of the study birds (p < 0.05). The current study observed a relationship between the different developmental stages of the parasite and immune responses ( humoral and lymphocytes responses). Broilers infected with sporulated oocysts (sporozoites) and merzoites yielded high CD4 T-lymphocyte numbers than the other groups, throughout the experiment periods (Tables 2.1, 2.2 and 2.3).
Figure 1.1: CD4 Levels in the plasma of broilers treated and non treated
infected with the developmental stages of the parasite at primary
infection
Figure 1.2: CD4 levels in the plasma of broilers treated and non treated
infected with the developmental stages of the parasite at secondary
infection.
Figure 1.3: CD4 levels in the plasma of broilers treated and non treated
infected with the developmental stages of the parasite at tertiary infection
Discussion
The cytokines generally reported in this study include IFN-
γ, IL-1, IL-2, IL-4, IL-6, TNF and TGF confirmed the findings of
Oldham and Gadde [14,15]. The study also showed that infection
of broilers with various developmental stages of Eimeria tenella
elicited both cellular. Homologous and heterologous challenges
of the birds with the various developmental stages of the
parasite at primary, secondary and tertiary infection levels
stimulated the secretion and proliferation of lymphocytes. This
is in concordance with the findings of Chapman, who reported
that the primary infection with Eimeria tenella oocyst induced
complete protection against homologous challenges [16]. The
findings from the study revealed the immunogenicity of the
developmental stages of the Eimeria tenella and this is consistent
with the reports of Molloy and Chow, who stated that the surface
antigens (SAGs) of the different Eimeria tenella stages were
Table 2.1: CD4 Lymphocytes subset counts (cells /103 μl) in experimental broilers orally infected with E. tenella developmental stages at primary infection treated and non treated
Group and |
Time (days) |
||||
2 |
4 |
6 |
8 |
10 |
|
I |
133.3 ± 58.6 |
158.3 ± 61.1 |
143.3 ± 57.7 |
133.3 ± 61.1 |
163.3 ± 57.7 |
II |
120.0 ± 65.6 |
180.0 ± 58.9 |
139.3 ± 57.7 |
131.3 ± 60.8 |
186.0 ± 85.4 |
III |
143.3 ± 84.7 |
147.3 ± 84.7 |
140.3 ± 85.8 |
133.3 ± 85.5 |
153.3 ± 76.4 |
IV |
126.7 ± 55.1 |
166.0 ± 50.7 |
110.7 ± 53.5 |
146.0 ± 63.9 |
183.7 ± 90.2 |
V |
144.0 ± 95.5 |
170.0 ± 85.4 |
146.7 ± 76.4 |
153.3 ± 68.1 |
180.0 ± 85.4 |
VI |
134.7 ± 102.5 |
198.0 ± 86.6 |
195.0 ± 86.6 |
198.0 ± 86.6 |
198.0 ± 86.6 |
Treated |
|
|
|
|
|
I |
198.7 ± 91.4 |
200.0 ± 117.9 |
160.0 ± 52.9 |
195.3 ± 95.1 |
199.0 ± 95.4 |
II |
146.0 ± 87.0 |
200.0 ± 86.8 |
162.7 ± 83.6 |
168.0 ± 95.4 |
199.0 ± 95.4 |
III |
150.0 ± 90.4 |
200.0 ± 86.6 |
160 ± 96.0 |
166.7 ± 83.3 |
198.0 ± 86.6 |
IV |
163.0 ± 88.9 |
198.7 ± 94.3 |
171.0 ± 86.6 |
177.0 ± 88.9 |
193.3 ± 90.7 |
V |
135.0 ± 91.7 |
200.0 ± 91.7 |
155.3 ± 96.5 |
169.7 ± 99.9 |
200.0 ± 86.6 |
No significant difference was observed across the days for both treated and non treated (p > 0.05). However, comparison of treated and non treated
showed significant difference (p < 0.05) for all the days.
Table 2.2: CD4 Lymphocytes subset counts (cells /103 μl) in experimental broilers orally infected with E. tenella developmental stages at primary infection treated and non treated
Non treated |
days |
||||
2 |
4 |
6 |
8 |
10 |
|
I |
140.0 ± 60.8 |
144.0 ± 62.9 |
140.0 ± 40.0 |
154.0 ± 64.1 |
156.7 ± 70.1 |
II |
183.3 ± 82.5 |
185.3 ± 86.0 |
187.3 ± 90.7 |
189.3 ± 95.0 |
192.7 ± 84.7 |
III |
143.3 ± 62.5 |
145.3 ± 63.6 |
146.7 ± 64.3 |
170.0 ± 50.7 |
160.0 ± 69.3 |
IV |
180.0 ± 85.4 |
168.0 ± 76.9 |
178.7 ± 81.0 |
177.3 ± 86.7 |
183.3 ± 85.8 |
V |
166.7 ± 76.4 |
170.7 ± 77.1 |
172.0 ± 76.9 |
166.0 ± 73.5 |
174.0 ± 78.1 |
VI |
183.3 ± 104.1 |
200.0 ± 86.6 |
200.0 ± 86.6 |
200.0 ± 86.6 |
200.0 ± 86.6 |
Treated |
|
|
|
|
|
I |
200.0 ± 86.6 |
196.0 ± 99.1 |
190.0 ± 96.4 |
199.3 ± 95.3 |
200.0 ± 86.6 |
II |
196.7 ± 95.2 |
188.7 ± 90.3 |
194.0 ± 91.2 |
197.3 ± 95.0 |
197.0 ± 96.0 |
III |
200.0 ± 86.6 |
192.7 ± 90.7 |
196.0 ± 92.6 |
200.0 ± 86.6 |
200.0 ± 86.6 |
IV |
193.3 ± 95.0 |
195.3 ± 95.1 |
196.0 ± 95.2 |
196.0 ± 91.1 |
198.7 ± 94.5 |
V |
180.0 ± 85.4 |
184.0 ± 85.9 |
186.0 ± 83.1 |
182.0 ± 81.6 |
187.3 ± 81.8 |
No significant difference was observed across the days for both treated and non treated groups (p > 0.05). However, comparison of treated and
non treated showed significant difference (p < 0.05) for all the days.
Table 2.3: CD4 Lymphocytes subset counts (cells /103 μl) in experimental broilers orally infected with E. tenella developmental stages at tertiary
infection treated and non treated
Group and |
Time (days) |
||||||
5 |
7 |
11 |
14 |
17 |
20 |
24 |
|
I |
133.3 ± 57.7 |
140.0 ± 60.8 |
140.0 ± 69.3 |
156.7 ± 67.0 |
160.7 ± 70.4 |
169.3 ± 76.8 |
173.3 ± 77.7 |
II |
193.3 ± 90.7 |
194.7 ± 95.6 |
194.7 ± 77.1 |
186.0 ± 88.5 |
194.0 ± 87.8 |
191.3 ± 93.6 |
194.3 ± 85.6 |
III |
165.3 ± 56.6 |
166.0 ± 28.6 |
134.0 ± 58.0 |
168.7 ± 46.3 |
138.7 ± 46.3 |
151.3 ± 66.9 |
166.7 ± 76.4 |
IV |
186.7 ± 90.2 |
180.7 ± 85.9 |
189.3 ± 82.4 |
174.3 ± 79.3 |
171.0 ± 78.6 |
174.0 ± 81.3 |
188.0 ± 90.3 |
V |
140.0 ± 60.8 |
146.7 ± 64.5 |
148.0 ± 64.5 |
171.3 ± 79.1 |
191.0 ± 41.4 |
174.0 ± 77.8 |
175.3 ± 81.1 |
VI |
200.0 ± 86.6 |
200.0 ± 86.6 |
200.0 ± 86.6 |
200.0 ± 86.6 |
200.0 ± 86.6 |
200.0 ± 86.6 |
200.0 ± 86.6 |
Treated |
|
|
|
|
|
|
|
I |
194.0 ± 95.5 |
195.3 ± 86.2 |
196.7 ± 95.2 |
198.0 ± 95.3 |
197.3 ± 98.0 |
192.7 ± 90.7 |
196.7 ± 89.5 |
II |
196.3 ± 87.4 |
199.3 ± 95.3 |
214.0 ± 68.2 |
196.0 ± 84.9 |
198.7 ± 90.8 |
197.3 ± 98.0 |
200.0 ± 86.6 |
III |
186.7 ± 81.4 |
178.7 ± 94.2 |
190.7 ± 88.9 |
194.0 ± 87.8 |
166.7 ± 76.4 |
165.3 ± 71.8 |
188.7 ± 90.3 |
IV |
194.0 ± 87.8 |
195.3 ± 95.1 |
198.0 ± 97.5 |
170.7 ± 77.1 |
180.7 ± 85.9 |
186.7 ± 81.4 |
190.7 ± 97.0 |
V |
182.0 ± 83.1 |
180.7 ± 85.9 |
179.3 ± 79.2 |
206.0 ± 45.9 |
184.7 ± 86.3 |
154.0 ± 66.9 |
188.7 ± 81.7 |
No significant difference was observed across the days for both treated and non treated (p > 0.05). A comparison of treated and non treated showed
significant difference (p < 0.05).
capable of initiating humoral and cytokines responses in birds
[17,18]. The study reported a strong immune responses (humoral
and cell mediated) in broilers infected with sporulated oocyst
(sporozoite) and merozoites. This is similar to finding of Rose
and Hesketh, Lillehoj, et al [18,19,20,21]. The strong immune
response can be attributed to the relative invasive features of
the parasite in nature due to proteins released from micronemes
which are important for host binding and invasion. The rhoptry
proteins secreted during invasion to form the parasitephorous
value within which the parasite resides. Also, the possession
of glicosulphosphatidylinositol (G.P.I) –linked surface antigens
(SAGs) may mediate binding to the host [22]. However, this study
is inconsistent with work of Onaga and Nakamiura [23].
The present study revealed an increase in the number of CD4 cells at day 10 post primary infection in both treated and non treated broilers as shown by Lillehoj [24]. Our study also recorded high numbers of CD4 cells after secondary infection of the birds at day 10 as against day 6 recorded by Lillehoj [24]. This difference may be due to differences in the age of the broilers, the strain of the parasite used or the genetic background of the birds [25]. The present study showed the expression of CD4 cells subset by blood lymphocytes of broilers infected with the various developmental stages of Eimeria tenella. This may indicate the induction of adaptive immune response to the infection [26]. The CD4 cell count expresses the numerical reactions of the broilers to oral administration of the different developmental stages of Eimeria tenella, revealing the stimulation of the immune system. This agrees with the reports of Hong and Lemus[27,28]. The study also revealed that the number of CD4 cells were significantly higher in the treated, than non-treated broilers. This is consistent to the finding of Hong, et al., who reported higher number of CD4 lymphocytes in infected birds treated than the non-treated ones [27].
The present study also demonstrated that the CD4 lymphocytes count increased at the different periods of infection with the various developmental stages of the parasite in both the treated and non- treated broilers. This is similar to the study of Bassey et al., who observed that the CD4 changes follow the phases of the parasite cycle for the Eimeria species considered [29]. There was also no significant difference in the CD4 cells count in both the treated and non-treated birds at secondary and tertiary infections. This is in accordance with the reports of Bassey et al. in infections of birds with Eimeria tenella [29]. In summary, this work add to our understanding of the ability of the various developmental stages of Eimeria tenella to induce immune responses in the chicken.
The present study revealed an increase in the number of CD4 cells at day 10 post primary infection in both treated and non treated broilers as shown by Lillehoj [24]. Our study also recorded high numbers of CD4 cells after secondary infection of the birds at day 10 as against day 6 recorded by Lillehoj [24]. This difference may be due to differences in the age of the broilers, the strain of the parasite used or the genetic background of the birds [25]. The present study showed the expression of CD4 cells subset by blood lymphocytes of broilers infected with the various developmental stages of Eimeria tenella. This may indicate the induction of adaptive immune response to the infection [26]. The CD4 cell count expresses the numerical reactions of the broilers to oral administration of the different developmental stages of Eimeria tenella, revealing the stimulation of the immune system. This agrees with the reports of Hong and Lemus[27,28]. The study also revealed that the number of CD4 cells were significantly higher in the treated, than non-treated broilers. This is consistent to the finding of Hong, et al., who reported higher number of CD4 lymphocytes in infected birds treated than the non-treated ones [27].
The present study also demonstrated that the CD4 lymphocytes count increased at the different periods of infection with the various developmental stages of the parasite in both the treated and non- treated broilers. This is similar to the study of Bassey et al., who observed that the CD4 changes follow the phases of the parasite cycle for the Eimeria species considered [29]. There was also no significant difference in the CD4 cells count in both the treated and non-treated birds at secondary and tertiary infections. This is in accordance with the reports of Bassey et al. in infections of birds with Eimeria tenella [29]. In summary, this work add to our understanding of the ability of the various developmental stages of Eimeria tenella to induce immune responses in the chicken.
Conclusion
The following can be concluded from the results obtained:
1. An immune response against ceacal coccidiosis could be established by immunization with Eimeria tenella-specific sporulated oocyst (sporozoites) and merozoites as well as other stages in birds of less than four weeks old
2. The prominent cytokines detected in the infected broilers were IFN- γ , IL-2, IL-4, IL-6, TNF and TGF, while the immunoglobulins are IgG or IgY.
3. Circulating CD4 lymphocytes subset count increased with the duration of infection.
1. An immune response against ceacal coccidiosis could be established by immunization with Eimeria tenella-specific sporulated oocyst (sporozoites) and merozoites as well as other stages in birds of less than four weeks old
2. The prominent cytokines detected in the infected broilers were IFN- γ , IL-2, IL-4, IL-6, TNF and TGF, while the immunoglobulins are IgG or IgY.
3. Circulating CD4 lymphocytes subset count increased with the duration of infection.
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