Kochs postulate in reproduction

of broiler coccidiosis by coinfection with eight most common

Eimeria spp.: a model for future
evaluation of new biologics

By:

Danielle B.
Ayyash

Purpose:
Establish a model of Kochs postulate for
reproducing coccidiosis in broilers
Using this model in future evaluation of
new controlling biologics

Experimental Design :
Table 1. Description of the different treatments of broilers
Groups of
Broilers

Number of Broilers

Broiler age at
Challenge (d)

Broiler age at
Sacrification (d)

10

14

20

10

21

27

10

28

34

10

35

41

10

20

10

27

10

34

10

41

Challenged

Control

Eimeria inocula
The esophageal Eimeria-challenge was with a
total of 1.76x105 oocysts/bird

E. acervulina, E. mivati, E. maxima, E.

necatrix, E. hagani, E. praecox, E. tenella,
and E. brunetti

Materials and Methods

Evaluation of the Kochs postulate during the 6 days incubation
period following an Eimeria challenge at assigned dates of 14,
21, 28 & 35 d of age
Protocols for Anti-coccidial Susceptibility Testing (AST)
1. The mean percent of weight increase
2. The mean feed conversion ratio
3. The mean percent mortality,

Materials and Methods

4. The mean score of lesions
The scoring of the lesions was accomplished by
a trained researcher, scoring 0 1, 2, and 3
(-ve, mild , moderate, severe )
5. The mean oocyst counts per gram of intestine
- 1cm of 4 different parts of intestine was rinsed in a saline
solution to remove fecal material
- Weighed
- homogenized and put in a 35% NaCl solution for the
floatation of oocytes.
- floated oocytes were counted in a McMaster chamber

Results & Discussion

Table 2. Percentage of weight increase in the four control and the four
Eimeria-challenged groups after 6 days of assigned challenges
Group1 of
broilers

Age at
assigned
challenge
(days)

Percentage of weight increase at 6 days

period following each assigned challenge
Controls
Challenged
SEM2

1
2
3
4
Mean

14
21
28
35

96.3
69.0
41.2
28.8
58.8a

88.2
54.5
31.7
20.2
48.6a

Each of the four control and four challenged groups had 10 birds
SEM is Standard Error of Means
a
Means in a row followed by the same alphabet superscript are not
significantly different (P>0.05)
1
2

10.0

Table 3. Feed conversion in the four control and the four Eimeriachallenged groups at 6 days-incubation period following the assigned
challenges
Group1 of
Age at
Feed conversion during 6 days
broilers
assigned
following challenge
challenge
Controls
Challenged
SEM2
(days)

1
2
3
4
Mean

14
21
28
35

1.1
1.7
2.1
2.1
1.7a

3.0
3.3
2.5
3.7
3.1b

0.3

Each of the four control and four challenged groups had 10 birds
SEM is Standard Error of Means
a,b
Means in a row followed by different alphabet superscripts are significantly different
(P<0.05)
1
2

Table 4. Frequency of mortality in the four control and the four Eimeriachallenged groups at 6 days period following the assigned challenges
Group1of
Age at assigned Frequency of mortality (%) in controls
broilers
challenge
and Eimeria-challenged birds at 6 days
(days)
period
following
the
assigned
challenges
Controls
Challenged SEM2
1
14
1 (10%)a
4 (40%)a
2
21
1 (10%)a
2 (20%)a
3
28
0 (0%)a
0 (0%)a
4
35
0 (0%)a
0 (0%)a
1
a
Each of the four control and the four Eimeria
challenged
groups
hada 10 0.5
birds.
Mean
0.5 (5%)
1.5
(15%)

SEM is Standard Error of Means

a
Frequencies in a row followed by the same superscript are not significantly different
(P>0.05)
2

Table 5. Mean score of lesions in different parts of the intestine in the four
control and the four Eimeria-challenged groups at the end of the 6 daysincubation period following the assigned challenges
Group1of
broilers

Age at assigned Intestinal part Mean score of lesions at 6 days period following
challenge
each challenge
(days)
Controls
Challenged
SEM2

14

21

28

35

Duodenum
Jejunum
Ileum
Cecum
Duodenum
Jejunum
Ileum
Cecum
Duodenum
Jejunum
Ileum
Cecum
Duodenum
Jejunum
Ileum
Cecum

0.0a
0.0a
0.0a
0.0a
0.2a
0.0a
0.1a
0.2a
0.6a
0.3a
0.0a
0.0a
0.25a
0.0a
0.0a
0.0a

1.5b
1.1b
1.6b
1.1b
2.0b
1.7b
1.6b
1.6b
1.4a
1.6b
0.6b
0.9b
0.5a
0.38a
0.38a
0.13a

0.25
0.18
0.25
0.21
0.27
0.25
0.26
0.25
0.25
0.25
0.15
0.22
0.18
0.14
0.14
0.06

Table 6. Mean oocysts count per gram of intestinal part in the four control
and the four Eimeria-challenged groups at the end of 6 days period
Group
of
broilers

Age at
assigned
challenge
(days)

Intestine
part

Mean oocyst count per gram of

intestinal organ at end of 6 days period
following the assigned challenges
Controls Challenged
SEM

14

21

28

35

Duodenum
Jejunum
Ileum
Cecum
Duodenum
Jejunum
Ileum
Cecum
Duodenum
Jejunum
Ileum
Cecum
Duodenum
Jejunum
Ileum

0a
0a
0a
0a
0a
0a
0a
0a
0a
0a
0a
0a
0a
0a
0a
a

42254b
67839b
112843b
96666b
33715b
36636b
41896b
48041b
6578b
12321b
19029b
7309b
15344a

6506b
2912a

9744
16177
35342
26708
8584
9343
10173
14776
1438
3075
4566
1978
6956
1569
921

In Conclusion
The Kochs Postulate for reproducing coccidiosis in broilers,
by the 8 most common Eimeria spp
- Significant statistical differences in
most measured parameters between
controls and challenged birds
- A trend of reduction in weight gain
and in augmentation of mortality in
challenged birds compared to controls.
Allow for future development of new immuno-modulators
and coccidiostats

Thank you
for your
Attention !