CHAPTER THREE
MATERIALS AND METHODS
3.1 Experimental
Animals:
The animals used for this experiment
were 90 day old Anak 2000 chicks, which
were obtained from a reputable dealer in Abakaliki.
3.2
Experimental Site
The experiment was carried out at Poultry
Unit of the Teaching and Research farm, Department of Animal Science, Ebonyi State
University Abakaliki, Nigeria. The
analyses were done at Department of Medical Labouratory Technology and Research Institute,
Ebonyi State University Abakaliki, Nigeria.
3.3
Experimental Diet.
Papaya leaves were harvested from
around the compounds and villages around Abakaliki town. They were dried at
room temperature then crushed to a finer state. The diet was formulated, using
locally sourced materials that were cheap and affordable. Table 1 shows the
percentage composition of the diet.
Table 1: The percentage composition of the
experimental starter diets (%).
|
|
TRT 1
|
TRT 2
|
TRT 3
|
TRT 4
|
TRT 5
|
|
Maize
|
50
|
50
|
50
|
50
|
50
|
|
Wheat offals
|
6
|
8
|
6
|
5
|
8
|
|
SBM
|
10
|
10
|
10
|
10
|
10
|
|
GNC
|
19
|
16
|
14
|
10
|
18
|
|
PKC
|
6
|
4
|
3
|
3
|
7
|
|
PLM
|
0
|
5
|
10
|
15
|
0
|
|
FISH
|
3
|
3
|
3
|
3
|
3
|
|
LIMESTONE
|
3
|
0
|
0
|
0
|
0
|
|
BONE MEAL
|
2
|
3
|
3
|
3
|
3
|
|
PRE MIX
|
0.25
|
0.25
|
0.25
|
0.25
|
0.25
|
|
SALT
|
0.25
|
0.25
|
0.25
|
0.25
|
0.25
|
|
LYSINE
|
0.25
|
0.25
|
0.25
|
0.25
|
0.25
|
|
METHIONIME
|
0.25
|
0.25
|
0.25
|
0.25
|
0.25
|
|
ORGACID
|
-
|
-
|
-
|
-
|
22g
|
|
TOTAL
|
100
|
100
|
100
|
100
|
100
|
|
ME(kg)
|
12.2
|
12.53
|
12.79
|
12.97
|
12.2
|
|
% CPV
|
22.12
|
22.15
|
22.2
|
22
|
21.7
|
|
% CF
|
4.2
|
4.3
|
4.2
|
4.2
|
4.4
|
TABLE 2: FINISHER MASH
|
TRT 1
|
TRT
1
|
TRT
2
|
TRT
3
|
TRT
4
|
TRT
5
|
|
|
Maize
|
52
|
52
|
52
|
52
|
52
|
|
|
Wheat
Offals
|
8
|
8
|
8
|
5
|
8
|
|
|
SBM
|
6
|
6
|
6
|
6
|
6
|
|
|
GNC
|
15
|
14
|
12
|
12
|
15
|
|
|
PKC
|
12
|
8
|
5
|
3
|
12
|
|
|
PLM
|
0
|
5
|
10
|
15
|
0
|
|
|
Fish
|
3
|
3
|
3
|
3
|
3
|
|
|
PLM
|
0
|
0
|
0
|
0
|
0
|
|
|
Limestone
|
0
|
0
|
0
|
0
|
0
|
|
|
Bone
meal
|
3
|
3
|
3
|
3
|
3
|
|
|
Premix
|
0.25
|
0.25
|
0.25
|
0.25
|
0.25
|
|
|
Salt
|
0.25
|
0.25
|
0.25
|
0.25
|
0.25
|
|
|
Lysine
|
0.25
|
0.25
|
0.25
|
0.25
|
0.25
|
|
|
Methionine
|
0.25
|
0.25
|
0.25
|
0.25
|
0.25
|
|
|
Orgacid
@ 1.5 kg/ton
|
|
|
45g
|
|
||
|
Total
|
100
|
100
|
100
|
100
|
100
|
|
|
ME
(kJ)
|
12.25
|
12.60
|
12.65
|
13.26
|
12.31
|
|
|
%
CPV
|
19.69
|
20.35
|
18.20
|
21.15
|
19.72
|
|
|
%
CF
|
4.16
|
4.12
|
3.32
|
3.75
|
4.25
|
|
3.
Experimental Design
The birds were randomly allocated to
five experimental diets namely; T1(control), T2(5% PLM), T3(10% PLM), T4(15%
PLM), and T5( 22g or orgacid).
3.4 Management
of the Animals.
The poultry unit was cleared of
cobwebs, swept and disinfected using diazintol (Diazinon dimphlate) and formaldehyde.
The pen was allowed to rest for 2 days. On arrival of the birds they were
randomly assigned to their replicates according to treatments used.
The cleaning of the house was repeated at
regular intervals till the end of the experiment. Water and feed troughs were
also cleaned on daily basis.
Table 3: The proximate analysis of the diets
Starter diets
|
|
%CP
|
%CF
|
ME(kg)
|
EE
|
|
T1
|
22.29
|
4.25
|
12.16
|
3.64
|
|
T2
|
22.32
|
4.31
|
12.51
|
3.72
|
|
T3
|
22.37
|
4.36
|
12.62
|
3.79
|
|
T4
|
22.20
|
4.21
|
12.98
|
3.88
|
|
T5
|
22.31
|
4.25
|
12.16
|
3.64
|
Table 4 Finisher diets
|
|
%CP
|
%CF
|
ME(kg)
|
EE
|
|
T1
|
19.69
|
4.16
|
12.25
|
3.67
|
|
T2
|
20.35
|
4.12
|
12.60
|
3.79
|
|
T3
|
18.20
|
3.32
|
12.65
|
3.85
|
|
T4
|
21.15
|
3.75
|
13.26
|
3.90
|
|
T5
|
19.72
|
4.25
|
12.31
|
4.25
|
3.6
Sample Collection
Faecal droppings were collected on
the Day 1 of the trial and weekly
thereafter till the last week of the experiment.
At the 8th
week 15 birds, one per replicate were slaughtered and the small intestines and
caecum taken for microbial count. The small intestine and caecum were
preserved on ice in a cold flask till it
reached the laboratory.
3.7 Materials Used in Culture
The media used were Eosinmethylene
blueagar (EMB) and Salmonella – Shigella agar (SSA). Eosin methylene blue Agar
(EMB) is used for the primary isolation of, and for differentiating between
enterobacteria. The media contains (W/V), peptone (1%), lactose and sucrose
(each at 0.5%), or lactose only at (1%), K2HPO4 (0.2%),
eosin Y (0.04%), methylene blue (0.0065%), agar (91.5%).
Salmonella –Shigella Agar (SSA)
contains Agar (13.5 g/l), lactose (10.0g/l), bite salts (8.5g/l), Na2S2O3
(8.5g/l), sodium citrate (8.5g/l), beef extract (5.0g/l), pancreatic digest of
casein (2.5g/l), peptic digest of animal tissue (2.5g/l).
All media were prepared according to
manufacturer’s instructions and were poured into a sterile culture plate and
allowed to gel.
Faecal (weekly)
droppings and the intestines and caecum from each representative repricate
(bird) were used.
3.8 Stages of the culture process.
3.8.1. Macroscopy:
This is the physical examination of the stool sample to ascertain is colour and
consistency.
Colour – whitish
/blackish
Consistency –
formed.
3.8.2. Preparation of Media
The media were
prepared according to the manufacturer’s instructions.
3.8.4 Inoculation
The last diluent was inoculated using 0.1
ml from the last test tube and inoculated using the media for the experiment
and the content was incubated for 24 hours.
3.8.5 Colony Count
The colonies were counted and
recorded, using light microscope to view the stained slides and data recorded
for each replicates.
3.9 Intestine and Caecum Content Culture.
0.5g each of digest from the small
intestinal or caecum was collected and
underwent the same procedure as in the
faecal culture. The results were recorded also.
3.10
Data Analysis.
The weekly faecal samples and
digesta from small intestinal and caecum
were subjected to One-Way Analysis of Variance (ANOVA). Where
differences existed,their means were separated using Fishers Least Significant
Difference (F-LSD) (One tail test) (Snedecor and cochran, 1980).
The linear
additive model for CRD experiment (Obi, 2002) is as follows:
Xij =
μ + ∑I + ∑ij
Where
Xij = individual observation
μ = population or
overall mean
∑I = treatment effect
∑ij = experimental error of
residual
CHAPTER FOUR
RESULT
4.1 THE MEAN COUNT (cfu/g) of Escherichia coli of BROILER DROPPINGS
The Result of
The Mean Count (cfu/g) of Escherichia
coli of Broiler Droppings as
presented in table 5.
TABLE
5: Mean Count (cfu/g) 0f Escherichia coli of broiler
droppings.
|
Escherichia Coli
|
T1
|
T2
|
T3
|
T4
|
T5
|
|
Week 1
|
6.4 x 105
|
7.2x 105
|
5.3 x 104
|
3.1 x 104
|
3.1 x 104
|
|
Week 2
|
4.2 x 104
|
3.7 x 104
|
6.4 x 104
|
5.6 x 104
|
1.6 x 104
|
|
Week 3
|
1.0 x 105
|
6.7 x 104
|
8.8 x 104
|
5.7 x 104
|
3.7 x 104
|
|
Week 4
|
9.3 x 105
|
3.2 x 104
|
6.7 x 104
|
4.4 x 104
|
3.1 x 104
|
|
Week 5
|
8.3 x 105
|
6.7 x 104
|
7.8 x 104
|
8.8 x 104
|
1.6 x 104
|
|
Week 6
|
1.0 x 105
|
6.2 x 104
|
4.1 x 104
|
4.1 x 104
|
1.0 x 104
|
|
Week 8
|
1.1 x 105
|
5.2 x 104
|
2.5 x 104
|
2.5 x 104
|
9.3 x 103
|
|
Mean
|
1.5
x 105
|
1.3
x 105
|
5.2
x 104
|
4.3
x 104
|
1.9
x 104
|
Mean count
SEM*
4.9 x
104
4.7 x 109 cfu/g. There were no
significant different (p>0.05) among the treatment
The
population of Escherichia coli is
highest on T2 (7.2 x 105 cfu/g) and least on the T4 (3.1 X 104cfu/g)
and T5 ( 3.1 x 104cfu/g) at week 1. Week 8 had the highest
population on T1 (1.5 x 105cfu/g) and least on T5 ( 1.9 x 104cfu/g).
4.2 THE
MEAN COUNT (cfu/g) of Salmonella
enteritidis of BROILER DROPPINGS.
The Result of The Mean Count (cfu/g) of Salmonella enteritidis of Broiler Droppings.
TABLE
6: Mean Count (cfu/g) of Salmonella enteritidis
of Broiler
Droppings.
|
Salmonella enteritidis
|
T1
|
T2
|
T3
|
T4
|
T5
|
|
Week 1
|
6.4 x 105
|
3.4x 104
|
2.9 x 104
|
4.4 x 104
|
2.9 x 104
|
|
Week 2
|
7.0 x 104
|
1.9 x 104
|
3.7 x 104
|
6.3 x 104
|
6.5 x 104
|
|
Week 3
|
2.7 x 104
|
2.5 x 104
|
3.1 x 104
|
2.0 x 104
|
6.8 x 104
|
|
Week 4
|
3.6 x 104
|
1.0 x 104
|
2.9 x 104
|
1.2 x 104
|
4.3 x 104
|
|
Week 5
|
6.4 x 104
|
2.2 x 104
|
1.5 x 104
|
2.5 x 104
|
1.2 x 104
|
|
Week 6
|
7.2 x 104
|
2.0 x 104
|
1.6 x 104
|
1.8 x 104
|
9.3 x 103
|
|
Week 8
|
7.8 x 104
|
1.5 x 104
|
9.3 x 103
|
1.3 x 104
|
1.0 x 104
|
|
Mean
|
1.1
x 105
|
1.8
x 104
|
2.1
x 104
|
2.4
x 104
|
2.1
x 104
|
Mean count
SEM*
2.4 x
104
6.7 x 108 cfulg. There were no
significant different (p>0.05).
The
population of Salmonella enteritidis is highest on T1 ( 6.4 X 105cfu/g) and
least on T3 and T4 ( 2.9 X 104cfu/g) at week 1.
4.3
THE MEAN COUNT (cfu/g) OF Shigella of BROILERS
DROPPINGS.
The Result of The mean count (cfu/g) of
Shigella of Broiler Droppings.
TABLE
7: Mean Count (cfu/g) of Shigella
of Broilers Droppings.
|
Shigella
|
T1
|
T2
|
T3
|
T4
|
T5
|
|
Week 1
|
8.3 x 104
|
3.1x 104
|
6.2 x 104
|
3.1 x 104
|
6.7 x 104
|
|
Week 2
|
1.3 x 105
|
8.8 x 104
|
9.9 x 104
|
7.2 x 104
|
6.2 x 104
|
|
Week 3
|
2.0 x 104
|
3.1 x 104
|
2.0 x 104
|
4.1 x 104
|
2.0 x 104
|
|
Week 4
|
3.6 x 104
|
3.1 x 104
|
2.0 x 104
|
3.1 x 104
|
2.0 x 104
|
|
Week 5
|
3.8 x 104
|
1.7 x 104
|
3.1 x 104
|
6.2 x 104
|
4.1 x 104
|
|
Week 6
|
9.3 x 104
|
1.7 x 104
|
2.0 x 104
|
3.1 x 104
|
2.4 x 103
|
|
Week 8
|
9.9 x 104
|
1.5 x 104
|
1.3 x 104
|
2.0 x 104
|
1.2 x 104
|
|
Mean
|
6.2
x 104
|
2.9
x 104
|
3.3
x 104
|
3.6
x 104
|
3.1
x 104
|
Mean count
SEM*
2.4 x
104
7.3 x 108 cfulg. There were no
significant (p>0.05) different among the means.
The population of Shigella is highest on T1 (
8.3 X 104cfu/g) and least on T2 ( 3.1 X 104cfu/g)
4.4
THE MEAN COUNT (cfu/g) of Lactobacillus spp of BROILER
DROPPINGS.
The Result of The Mean of COUNT (cfu/g) of Lactobacillus spp of Broilers Droppings.
TABLE
8: Mean Count (cfu/g) of Lactobacillus
spp of broilers
droppings.
|
lactobacillus spp
|
T1
|
T2
|
T3
|
T4
|
T5
|
|
Week 1
|
6.7 x 104
|
7.8x 104
|
1.0 x 105
|
9.3 x 104
|
1.0 x 105
|
|
Week 2
|
8.2 x 105
|
1.3 x 105
|
1.2 x 105
|
1.2 x 105
|
1.3 x 105
|
|
Week 3
|
8.2 x 104
|
2.1 x 105
|
2.1 x 105
|
2.6 x 105
|
1.3 x 105
|
|
Week 4
|
8.6 x 104
|
1.6 x 105
|
2.1 x 105
|
2.6 x 105
|
2.5 x 105
|
|
Week 5
|
7.9 x 104
|
2.1 x 105
|
2.5 x 105
|
2.5 x 105
|
2.6 x 105
|
|
Week 6
|
8.9 x 104
|
2.0 x 105
|
2.1 x 105
|
2.5 x 105
|
2.0 x 104
|
|
Week 8
|
9.3 x 104
|
1.9 x 105
|
2.3 x 105
|
1.6 x 105
|
1.6 x 105
|
|
Mean
|
7.2
x 104
|
1.5
x 105
|
1.7
x 105
|
1.7
x 105
|
1.3
x 105
|
Mean count
SEM*
8.6 x
104
8.7 x 108 cfulg. There were no
significant (p>0.05) different at all treatments.
The mean count (cfu/g) of Lactobacillus spp of broilers droppings
were found to be no significant at (p>0.05) for all treatments considered.
The mean count (cfu/g) of Lactobacillus
spp of broilers droppings showed the highest count at T3 (10% PLM) and T4
(15% PLM) respectively. While the least mean count of Lactobacillus spp showed at T1(control).
4.5
THE MEAN COUNT (cfu/g0 of Staphylococcus
aureus of BROILER DROPPINGS.
The
Result of The Mean Count (cfu/g) of Staphylococcus
aureus Broiler Droppings.
TABLE
9: Mean Count (cfu/g) of Staphylococous
aureus of broilers
droppings.
|
Staphylococous aureus
|
T1
|
T2
|
T3
|
T4
|
T5
|
|
Week 1
|
5.2 x 104
|
4.9 x 104
|
1.2 x 105
|
5.0 x 104
|
8.3 x 104
|
|
Week 2
|
6.2 x 104
|
4.2 x 104
|
3.0 x 104
|
3.2 x 104
|
7.4 x 104
|
|
Week 3
|
7.4 x 104
|
1.0 x 104
|
1.3 x 105
|
1.6 x 105
|
1.2 x 105
|
|
Week 4
|
7.8 x 104
|
1.6 x 105
|
1.3 x 105
|
1.8 x 105
|
2.1 x 105
|
|
Week 5
|
8.4 x 104
|
1.6 x 105
|
2.1 x 105
|
1.7 x 105
|
1.7 x 105
|
|
Week 6
|
9.2 x 104
|
1.6 x 105
|
1.8 x 105
|
1.8 x 105
|
1.6 x 104
|
|
Week 8
|
8.8 x 104
|
1.6 x 105
|
2.4 x 105
|
1.4 x 105
|
1.9 x 105
|
|
Mean
|
6.6
x 104
|
9.3
x 104
|
1.3
x 105
|
1.1
x 105
|
1.3
x 105
|
Mean count
SEM*
6.6 x
104
6.3 x 109 cfulg. There were
significant different (p>0.05) at the means.
The mean count (cfu/g) of Staphylococcus aureus of broilers
droppings was found to be significant different at (p>0.05) for the means
considered. The mean count showed the highest value at T3(10% PLM) and T5 (22g
of orgacid) of Staphylococcus aureus of broiler droppings. The least mean count was
showed at T1(control) and T2 (5% PLM).
4.6
THE MEAN COUNT (cfu/g) of Lactobacillus
spp and THE ENTEROPATHOGENS in THE DIGESTA of BROILERS CAECUM and SMALL
INTESTINE at WEEK 8 of AGE.
The
Result of Lactobacillus spp and The Enteropathogens Mean Counts in The
Digesta of Broilers Caecum and Small Intestine at Week 8 of Age.
TABLE
10: Mean Count (cfu/g) of Lactobacillus
spp and the enteropathogens in the
digesta of broilers caecum and small intestine at week 8 of age.
|
Caecum
microbes
|
T1
|
T2
|
T3
|
T4
|
T5
|
|
Esotherichia coli
|
1.1 x 105
|
5.2 x 104
|
2.0 x 104
|
4.6 x 104
|
4.1 x 104
|
|
Salmmomella enteristidis
|
4.1 x 104
|
4.6 x 104
|
3.1 x 104
|
3.6 x 104
|
4.1 x 104
|
|
Shigella
|
7.8 x 104
|
3.6 x 104
|
2.5 x 104
|
3.1 x 104
|
3.1 x 104
|
|
Staphylococcus
aureus
|
8.8 x 104
|
1.8 x 105
|
2.0 x 105
|
1.6 x 105
|
1.5 x 105
|
|
Lactobacillus
spp
|
8.8 x 104
|
2.0 x 105
|
2.1 x 105
|
2.1 x 105
|
1.9 x 105
|
|
SEM
|
5.7 x 104
|
9.2 x 105
|
9.8 x 104
|
8.8 x 104
|
7.9 x 105
|
|
Small intestine microbes
|
|
|
|
|
|
|
Escherichia
coli
|
7.8 x 104
|
3.6 x 104
|
3.8 x 104
|
2.3 x 104
|
3.1 x 104
|
|
Salmonella
enteritidis
|
3.6 x 104
|
3.6 x 104
|
1.5 x 104
|
2.5 x 104
|
2.5 x 104
|
|
Shigella
|
5.7 x 104
|
2.5 x 104
|
2.5 x 104
|
2.0 x 104
|
2.5 x 104
|
|
Staphylococcus
aureus
|
3.6 x 104
|
7.2 x 104
|
9.3 x 104
|
1.0 x 105
|
6.2 x 104
|
|
Lactobacillus
spp.
|
4.6 x 104
|
9.9 x 104
|
2.1 x 105
|
1.4 x 105
|
1.1 x 105
|
There is significant different
(p<0.05)
The mean count of Lactobacillus spp and the enteropathogens in the digesta of
broilers caecum showed the highest mean count at T3( 10% PLM) and T4 (15% PLM)
respectively. While the least mean count of Lactobacillus
showed at T1 (control). The enteropathogens in the digesta showed that at
caecum the highest value were recorded at T3(10%PLM) showed by Staphylococcus aureus. While the least
count of enteropathogens were recorded at T3 (10% PLM) showed by Escherichia
coli.
The mean count of Lactobacillus spp and enteropathogens in the digesta of broilers
small intestine showed the highest mean count at T3(10% PLM). While the least
mean count of Lactobacillus spp showed at T1(control).
The enteropathogens in the digesta showed that at small intestine the highest
value were recorded at T4(15% PLM) showed by Staphlococcus aureus . While the least count of enteropathogens in
the small intestine were recorded at T3(10% PLM) showed by Salmonella enteritidis followed
by T4(15% PLM) showed by Shigella.
CHAPTER FIVE
5.0
DISCUSSION
In table 5, Escherichia coli in broilers droppings
were found to be no significant different (p>0.05). The mean count (cfu/g)
of Escherichia coli of broilers
droppings showed the highest mean value at T1(control)( 1.5x 105cfu/g)
followed by T2 (5% PLM) (1.3 x 105cfu/g). While those on T5(orgacid)
showed the least mean count (1.9 x 104cfu/g). This could be as
result of the effect of papaya leaf meal (PLM) which may be resistant to Escherichia coli. This is in line with
Mohanery and Mahzorieh (2005) stated a decrease in Escheichia coli population in the intestines of broiler chicken
with malic acid.
The mean count
(cfu/g) of Salmonella enteritidis of
broiler droppings was found to be no
significant (p>0.05) different. The result showed the highest mean count of Salmonella enteritidis at T1(control)
(1.1 x 105cfu/g) followed by T4 (15% PLM) (2.4 x 104+cfu/g).
Showed the least mean count at T2 (5% PLM) (1.8 x 104cfu/g) of Salmonella enteritidis in broiler
droppings. Which is agreement with (Krishna et al., 2008) which states that herbal containing papaya leaf meal (PLM) or
roots as one of the constituents has antibacterial activity against Salmonella tyhii, Salmonella enteritidis, etc.
The mean count (cfu/g)
of Shigella in broiler droppings were
found to be no significant (p>0.05) different. The result showed that at T1(control)
( 6.2 x 104cfu/g) has the highest value followed by T4(15% PLM) (3.6
x 104cfu/g) of Shigella in broiler droppings. While the
least count was seen at T2 (5% PLM)(2.9 X 104cfu/g) of Shigella in broiler droppings. This
agreed with (Nwofia et al., 2002) which said that purified extracts from
matured papaya leaf also produces very significant antibacterial activity on Staphlococcus aureus, Bacillus cereus,
Escherichia coli and Shigella
flexneri.
In table 8, the
result reviewed the mean count (cfu/g) of Lactobacillus
spp in broiler droppings. It showed that at T3(10% PLM) (1.7 X 105cfu/g) and T4(15% PLM) (1.7 X 105cfu/g)
had the highest mean count of Lactobacillus
spp in broiler droppings. While the least mean count was seen at
T1(control)( 7.2 x 104cfu/g) of Lactobacillus
spp in broiler droppings. This agreed with
Amit-Romach et al., (2004) that the predominant population of Lactobacillus spp as a result of
probiotic effect . This showed thath there were increase in Lactobacillus species more at T3(10%
PLM) and (T4 (15% PLM) as a result of the effect of papaya leaf meal (PLM) fed
to the birds.
The result showed on table 9 reviewed that
the mean count (cfu/g) of Staphylococcus
aureus of broilers droppings were found to be significant different
(p<0.05). The mean count showed the highest value of T3(10% PLM)(1.3 X 105cfu/g)
and T5(orgacid)(1.3 x 105cfu/g) respectively of staphylococcus aureus
in broilers droppings. The least mean count showed at T1(control) (6.6 x 104cfu/g)
and T2(5% PLM) (9.3 X 104cfu/g). This is also in agreement with
Nwofia et al, (2002) purified extracts from matured papaya leaf also produces
very significant antibacterial activity on Staphylococcus
aureus, Bacillus cereus, Escherichia coli.
The mean count of Lactobacillus
spp and the enteropathogens in the digesta of broilers caecum showed the
highest mean count at T3(10% PLM) (2.1 x 105cfu/g) and T4 (15% PLM)
(2.1 x 105cfu/g)
respectively. While the least mean count of Lactobacillus
spp in broiler droppings was seen at
T1(control) (8.8 x 104cfu/g). The enteropathogens in the digesta
showed that at caecum the highest value was recorded at T3(10% PLM) (2.0 x 105cfu/g)
showed by Staphylococcus aurues.
While the least count of enteropathogens was showed by Escherichia coli at T3(10%
PLM) (2.0 x104cfu/g). These result corroborate with soumaya et al, (2011) and Barnes et
al, (1972) that in the caecum the effect of lactic acid increased thus
decreasing the pathogenic baterial in the caecum.
The mean count of Lactobacillus spp and the
enteropathogens in the digesta of broilers small intestine showed that there
was an increased at T3(10% PLM) (2.1 x 105cfu/g). Showing a least
mean count of Lactobacillus spp in the digesta of broilers at T1( control)
(2.0 x 104cfu/g). The enteropathogens in the highest value were
recorded at T4 (15% PLM) ( 1.0 x 105cfu/g) showed by Staphylococcus aureus. While the least
mean count recorded at T3(10% PLM) (1.5 x 104cfu/g) showed by Salmonella enteritidis followed by T4(15% PLM) (2.0 x 104cfu/g)
showed by Shigella. This corroborated
with Brzokal et al,(
2007) microbiology analyses of the small intestine digesta showed that
lactic acid bacteria increased the intestinal counts of Lactobacillus spp compared
with control group and with antibiotic group.
5.1
Conclusion
The
research showed that in the digesta of broilers the mean count of Lactobacillus
spp increased significantly (P < 0.05) with both T3 and T4 having the
highest mean count of 1.7 x 105 cfu/g.The
population of Lactobacillus spp was observed to increase in T4(15% PLM) at
week 4 having a mean count of 1.7 x 104 cfu/g; and that of T5(
orgacid) at week 5 having a mean count of 2.6 x 105 .
Therefore
from all indication T3 (10% PLM) had the best lactobacillus spp count.
It is also observed that T3 and T4 had the lowest mean counts of enteropathogens
examined. Therefore if at T3 (10% PLM) and T4 (15% PLM) that it does not have
any physiological implication on the broilers then in summary, T3 (10% papaya
leaf meal) and T4 (15% papaya leaf meal (PLM) is the best level of inclusion.
5.
2 Recommendation
It
is therefore recommended that at T3 (10% PLM) and T4 (15% PLM) has the best microbicidal property. It can be
use since the lowest mean count of bacteria was found in them and also the
population of lactobacillus spp increased significantly in the same inclusion
levels.
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Serial Dilution
Procedure:
·
Five
test tube of equal measurement was set on a test tube rack.
·
5
mls of normal saline solution was added in the first test tube, that is, the
stock and 4 mls of normal saline was added in the subsequent four test tubes.
·
0.5g
of the sample and latter digesta from
measured with automatic measuring device.
·
The 5 mls of the normal saline was added to the
sample and was thoroughly mixed.
·
1
ml of the solution from the stock was transferred to the next test tube that
contain 4 mls of normal saline solution.
·
The
2nd test tube was shaken thoroughly 2 mins
and then 1 ml was collected from
it and added to the 3rd test tube.
·
The mixture in the 3rd test tube was
thoroughly shaken for 2 mins then 1ml was collected from it and placed in the 4th
test tube. The process was repeated also in the fourth and fifth test tubes.