The influence of drugs with a synergistic
mixture of phencides on the intensity of
invasion, morphological blood parameters and
leukocyte formula in chicken eimeriosis mixed
with colibacillosis
Feruza
Ibragimova
1*
,
Davletbay
Ibragimov
1
,
Asadullo
Daminov
1
, and
Gulmurod
Mamadullaev
2
1
Samarkand State University of Veterinary Medicine, Livestock and biotechnologies, Samarkand,
Uzbekistan
2
Veterinary Research Institute, Samarkand, Uzbekistan
Abstract.
In this work, we studied the synergistic preparations phencid
and phencid premix prepared by us in doses of 288 mg/kg and 1000 mg/kg
to the feed, which reduce the intensity of invasion by almost 4-5 times and
their use at the above doses does not negatively affect the morphological
parameters and the leukocyte formula of the blood of birds. Considering
the relevance, these drugs can be used for the prevention and treatment of
these diseases. For this purpose, birds were experimentally infected
simultaneously with pathogens of Eimeria and Escherichiosis. As a result,
it was found that the tested drugs reduce the intensity of invasion by 4-5
times and do not have a negative effect on the morphological composition
of the blood and the leukocyte formula.
1 Introduction
In the Republic of Uzbekistan, chicken breeding is one of the leading branches of livestock
breeding, and they are mainly raised in joint-stock companies, limited liability companies,
peasant farmers and private auxiliary farms. especially with colibacteriosis, most of the
chickens die, and the rest lag behind in growth and development, causing great economic
damage to farms. multiplying in the mucous membranes in a schizogonal way, it destroys
the integrity of the epithelial cells and opens the way for intestinal microorganisms to enter
the blood. At this time, inflammatory processes increase and prevent the absorption of
substances necessary for the div. For the prevention and treatment of these diseases,
coccidiostats and antibiotics are used separately [1-3, 5]. However, pathogens quickly get
used to the drugs used. Taking into account the above problems, an antibiotic, vitamin
phencid synergistic mixture and its phencid premix types were created on the basis of
ferulene isolated from the plant Ferula assofoetida, which grows in the mountainous and
*
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© The Authors, published by EDP Sciences. This is an open access article distributed under the terms of the Creative
Commons Attribution License 4.0 (https://creativecommons.org/licenses/by/4.0/).
desert regions of the Republic of Uzbekistan. Its effectiveness against colibacteriosis was
tested in laboratory experiments. We set ourselves the goal of studying the effect of the
drugs used in this experiment on the intensity of invasion and on the morphological
parameters of blood and leukocyte formula during a period of coexistence with Eimerioz
colibacterosis.
2 Materials and methods
For laboratory experiments, one-day-old chicks belonging to the "Ross 308" cross were
brought from the chicken farm and placed in a small chicken house in a common gala on
the beds, and at the age of fourteen days, five groups of 20 heads were formed. In
particular: the first is a relatively clean control group, they were fed with clean food without
drugs until the end of the experiment, the second is a control group infected with pathogens
and not treated, which were previously titrated sporulated oocysts with a killing dose of
O'D50-70 ( 0.5 ml of E. servulina 125 000, E. maxima 8000, E. tenella 25 000) in 1 mm
3
suspension was injected into the scrotum of each chick through a syringe probe, and E. coli
culture was injected into the abdomen by 0.3 ml. was sent to space. The number of oocysts
in the litter on the 5th, 7th, 10th, 15th and 20th days of the experiment according to the
GOST 25383-82 (ST SEB 2547-80) "Metody laboratornoy diagnostiki coccidiosis" manual.
is multiplied. Chicks in the third experimental group were given 288 mg/kg of the drug
with synergistic mixture of Phencid for 10-12 days immediately after infection. The
composition of the Fensid synergistic mixture drug is as follows: Ferulen 30mg/kg,
Levomycetin 188mg/kg and Phtalozol 70mg/kg were given with a total of 288mg/kg of
food. The chickens of the fourth experimental group were infected with eimeria and
colibacteriosis cultures and were given 1000 mg/kg of the drug in the form of a premix of
fencid for 10-12 days. Its composition is ferulene 30mg/kg, levomycetin 188mg/kg and
phthalozol 70mg/kg, vitamin U(S-methylmethionine) 0.003mg/kg, vitamin K 0.4mg/kg, the
rest is made up of santhoxin and wheat bran and chicks of group 5 were given Spektril-S
with 1 ml/2 l of water for 5 days, Spektrazuril-T with 1 ml/1 l of water for 2 days with
water throughout the day according to the instructions. During the experiment, blood was
taken from the underwing vein and the amount of hemoglobin in it was determined by the
hemoglobin-cyanide method (with acetoncyanhydrin) on the KFK-2 device according to
the instructions of [11] of et.all. The number of erythrocytes, leukocytes and thrombocytes
in 1 mm3 of blood was determined by counting with the method of [4] on the Goryayev
counting grid, stained with Romanov Giemza and methylviolet dyes. To determine the
leukocyte formula in the blood smear, it was stained by the Poppenheim method and
counted by the Filipchenko three-field method. The obtained numbers were subjected to
statistical processing according to the method of [10], and the degree of accuracy was
determined from the Student table. P<0.05.
The reproduction of oocysts is of great importance in the specific activity of the
synergistic mixture drugs used in the treatment of eimeriosis and colibacteriosis of chickens
and in the formation of immunity formed in the div against eimeriosis.
3 Results
Considering the above problems, it is of practical and scientific interest to determine the
spheres of influence of synergistic preparations of mixtures on the reproductive properties
of oocysts. To implement the task, on days 5-7-10-15 and 20 of the experiment, droppings
samples were taken from each group and the number of oocysts in 1 gram of droppings was
determined. The results of laboratory experiments are presented in Table 1.
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Table 1.
The influence of phencid and phencid premix on the intensity of invasion.
S/n
Group name
The drug
name
Dose (mg/kg
food)
Number
of
chicks
Save
flexible
rate (%)
Infestation intensity (1000
oocysts in 1 g of feces)
Inspection days
5
7
10 15 20
1
A relatively clean
control
-
-
20
100.0
-
-
-
-
-
2
Comparative
infected control
-
-
20
15.0
1.314 1.820 272 53 8
3
Experience
Phencid
288
20
100.0
263
524
61
8
1
4
Experience
Phencid
premix
1000
20
100.0
250
498
55
7
1
5
Experience
Spectril-S
Spectra-
zuril-T
5 days with
1ml/2l water 2
days with 1 ml/l
of water
20
100.0
301
544
69 57 2
During the experimental period, group 2 and group 5 chicks shed permanent oocysts
with their droppings. In particular, on the fiveth day of the experiment, chicks in the
untreated control group were exposed to 1,314 oocysts per 1 g of feces. The number of
oocysts in 1 gram of droppings was 263,000 when the chickens in the third experimental
group received the fencid synergistic mixture drug at 288 mg/kg of feed for 10 days, and
250,000 when the chicks in the fourth experimental group received the phencid premix
drug at 1000 mg/kg of feed for 10 days. and when chickens in the fifth experimental group
were given Spektril-S 1 ml/2l for 5 days, Spektra zuril-T 1 ml/l water for 2 days, the
number of oocysts in 1 g of litter was 301 thousand.
On the seventh day of the experiment, the chicks of the second group with 1 gram of
litter averaged 1 million 820,000, the chicks of the third experimental group with 1 gram of
litter 524 thousand, the chicks of the fourth experimental group with 1 gram of litter, and
the chicks of the fifth experimental group with 1 gram of litter 544 isolated a thousand
oocysts.
By the tenth day of the experiment, chicks of the second group released an average of
272,000 oocysts with 1 gram of feces, chicks of the third experimental group released
61,000 oocysts with 1 gram of feces, chicks of the fourth and fifth experimental groups
released 55-69 thousand pieces of oocysts with 1 gram of feces. 15-20 days of the
experiment by the end of the day, 53-8 thousand chicks of the second group, 8-1, 7-1
thousand and 57-2 thousand chicks of the 3rd, 4th and 5th experimental groups were
separated from oocysts with 1 g of feces. As a result of the obtained laboratory
experiments, chickens in the second comparative infected and untreated control group shed
1314-1820 thousand oocysts in 1 gram of feces on the 5th-7th day of the experiment.
Chicks in the remaining experimental group also secreted maximum oocysts with 1 g of
droppings. The phencid and phencid premix preparations used in the experiment released
several times less oocysts compared to the number of oocysts in the droppings of chicks of
the second group. This protects it from the damage of the external environment, even if
only partially.
Indicators indicating the general functional state of the div include indicators of
general hematological and serum proteins in the blood of chickens.
On days 5-7-10-15-20 of the experiment, blood was taken from the chickens from the
subwing vein, the amount of hemoglobin, the number of erythrocytes, leukocytes and
platelets in 1 mm3 of blood was determined, as well as the results obtained. when a blood
smear was prepared and the leukocyte formula was determined on these days are given in
tables 2-10.
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Table 2.
The influence of phencid and phencid-premix on the number of erythrocytes (10
12
/l) with a
mixed course of eimeria and colibacillosis in chickens M±m.
No.
Groups
Inspection days
5
7
10
15
20
1
Pure control group
2.61±0.14
2.55±0.14
2.67±0.09
2.70±0.08
2.68±0.07
2
Infected experimental group
1.88±0.16
х
1.32±0.06
ххх
2.50±0.17
2.71±0.09
2.71±0.06
3
The experimental group
phencid
2.54±0.09
2.56±0.07
2.63±0.09
2.69±0.20
2.63±0.28
4
The experimental group
phencid premix
2.59±0.11
2.58±0.09
2.64±0.12
2.72±0.15
2.67±0.05
5
Experimental group Spectril-
S Spectrazuril-T
2.60±0.15
2.57±0.05
2.58±0.15
2.71±0.19
2.69±0.14
Note: x-p<0.05; xxx-p<0.01
Table 3.
Effect of phencid and phencid premix on the amount of hemoglobin in the blood (g/l) with a
mixed course of eimeria and colibacillosis in chickens M±m.
No.
Groups
Inspection days
5
7
10
15
20
1
Pure control group
89.6±0.6
90.0±0.8
93.5±0.7
96.3±0.7
95.4±0.9
2
Infected experimental group
72.2±2.5
ххх
61.0±1.8
хххх
82.4±1.2
92.5±1.7
96.0±1.1
3
The experimental group phencid
89.0±0.2
87.5±0.9
88.9±0.9
95.5±0.3
95.8±0.7
4
The experimental group phencid
premix
87.4±0.9
89.8±0.92
90.0±0.8
97.0±0.5
96.8±0.6
5
Experimental group Spectril – S
Spectrazuril - T
88.5±0.8
89.0±1.0
89.4±0.7
95.7±1.5
94.3±1.5
Note:
xxx
-p<0.011;
хххх
-p<0.001
Table 4.
Effect of phencid and phencid premix on the number of leukocytes in the blood (10
9/l
) with a
mixed course of eimeria and colibacillosis in chickens M±m.
No.
Groups
Inspection days
5
7
10
15
20
1
Pure control group
26.4±0.41
27.0±0.51
27.0±0.73
26.7±0.50 27.1±0.49
2
Infected experimental group
27.1±1.48
29.4±1.64
х
38.4±2.0
ххх
32.2±1.52
х
27.4±1.27
3
The experimental group phencid
27.0±0.67
26.6±0.37
26.8±0.66
26.8±0.37 27.0±0.71
4
The experimental group phencid
premix
26.6±0.51
26.5±0.44
26.7±0.92
26.9±0.61 27.5±0.51
5
Experimental group Spectril-S
Spectrazuril-T
26.2±0.45
26.7±0.55
26.9±1.00
27.0±0.72 27.8±0.49
Note:
x
-p<0,05;
ххх
-p<0,01
Table 5.
The influence of phencid and phencid premix on the number of platelets (10
9/l
) with a mixed
course of eimeria and colibacillosis in chickens M±m.
No.
Groups
Inspection days
5
7
10
15
20
1
Pure control group
36.9±0.11 36.0±0.35 36.5±0.43 35.7±0.35 34.3±1.02
2
Infected experimental group
27.3±1.36
х
38.9±0.71
49.1±0.52
х
41.1±1.01
36.3±1.20
3
The experimental group phencid
36.7±0.51 35.0±2.03 36.9±0.63 36.1±1.09 36.0±1.02
4
The experimental group phencid premix 36.4±0.58 35.3±0.89 36.4±0.94 36.0±0.66 35.9±1.07
5
Experimental group Spectril-S
Spectrazuril-T
36.3±0.15 35.7±0.12 35.8±0.45 35.6±0.45 34.7±0.98
Note:
x
-p<0.05
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Table 6.
The influence of phencid and phencid premix on the number of basophils in the blood (%)
with a mixed course of eimeria and colibacillosis in chickens M±m.
No.
Groups
Inspection days
5
7
10
15
20
1
Pure control group
2.2±0.24 2.4±0.21 2.2±0.23 2.2±0.20 2.2±0.20
2
Infected experimental group
2.0±0.31
2.2±0.37
2.2±0.37
2.2±0.20
2.2±0.20
3
The experimental group phencid
2.0±0.31 2.2±0.37 2.2±0.37 2.2±0.20 2.2±0.20
4
The experimental group phencid premix
2.0±0.31 2.2±0.37 2.2±0.20 2.2±0.20 2.2±0.20
5
Experimental group Spectril-S Spectrazuril-T
2.2±0.24 2.2±0.21 2.2±0.23 2.2±0.21 2.2±0.23
Note:
x
-p<0.05
Table 7.
Effect of phencid and phencid premix on the number of eosinophils in the blood (%) in
mixed cases of eimeria and colibacillosis in chickens M±m.
No.
Groups
Inspection days
5
7
10
15
20
1
Pure control group
3.2±0.21
3.0±0.31
3.4±0.24 3.4±0.24 3.4±0.24
2
Infected experimental group
4.2±0.73
х
4.6±0.68
хх
3.2±0.20
3.2±0.21
3.4±0.24
3
The experimental group phencid
3.0±0.31
3.2±0.20
3.4±0.24 3.2±0.20 3.2±0.20
4
The experimental group phencid premix
3.0±0.31
3.2±0.37
3.2±0.20 3.2±0.20 3.4±0.24
5
Experimental group Spectril-S
Spectrazuril-T
3.2±0.21
3.2±0.20
3.2±0.24 3.2±0.21 3.4±0.20
Note:
x
-p<0.05;
хх
-p< p<0.02
Table 8.
The influence of phencid and phencid premix on the number of pseudoeosinophils in the
blood (%) in a mixed course of chicken eimeria and colibacillosis M±m.
No.
Groups
Inspection days
5
7
10
15
20
1
Pure control group
28.8±0.88
28.2±0.57
27.8±0.49
26.8±1.76 27.6±1.80
2
Infected experimental group
44.4±1.03
х
46.8±2.49
хх
39.2±1.24
х
30.8±1.52
27.8±1.88
3
The experimental group phencid
28.6±0.33
28.4±0.45
28.0±1.73
27.6±1.40 27.4±0.92
4
The experimental group phencid
premix
27.4±1.43
28.2±1.65
26.8±1.56
26.6±0.71 27.2±1.02
5
Experimental group Spectril-S
Spectrazuril-T
27.6±0.35
28.5±0.52
27.2±0.71
27.1±0.35 27.8±1.22
Note:
x
-p<0.05
Table 9.
The influence of phencid and phencid premix on the number of lymphocytes in the blood
(%) with a mixed course of eimeria and colibacillosis in chickens M±m.
No.
Groups
Inspection days
5
7
10
15
20
1
Pure control group
58.0±0.94
56.8±2.99 57.0±2.06 56.6±1.20 56.2±1.15
2
Infected experimental group
49.2±2.15
х
39.2±1.62
хх
42.0±2.60
х
48.8±1.98
56.8±1.06
3
The experimental group phencid
58.4±0.67
57.0±2.58 57.6±2.29 57.4±1.43 57.4±1.20
4
The experimental group phencid
premix
57.2±1.11
57.2±2.17 58.4±1.99 56.2±2.03 57.6±0.81
5
Experimental group Spectril-S
Spectrazuril-T
57.6±0.87
57.3±2.71 57.6±2.55 56.9±1.49 57.5±0.92
Note:
x
-p<0.05;
хх
-p< 0.02
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Table 10.
The influence of phencid and phencid premix on the number of monocytes in the blood (%)
with a mixed course of eimeria and colibacillosis in chickens M±m.
No.
Groups
Inspection days
5
7
10
15
20
1
Pure control group
4.6±0.51
4.4±0.51 4.4±1.20 4.6±0.81
4.6±0.60
2
Infected experimental group
6.6±0.92
х
6.6±1.21
х
7.2±0.58
х
5.4±0.51
4.8±0.37
3
The experimental group phencid
4.6±0.51
4.6±0.51 4.4±0.67 4.6±0.81
4.4±0.40
4
The experimental group phencid premix
4.6±0.51
4.4±0.81 4.4±0.51 4.0±0.70
4.6±0.51
5
Experimental group Spectril-S
Spectrazuril-T
4.6±0.55
4.4±0.27 4.4±0.37 4.4±0.45
4.6±0.49
Note:
x
-p<0.05
The main changes in blood composition were observed in the blood parameters and
leukocyte formula of chickens of the control group that were not exposed to secondary
infection. However, the blood parameters of chickens from the experimental groups did not
differ from the blood parameters and leukocyte formula of chickens from the pure control
group.
Thus, on the 5th day of the experiment, the number of erythrocytes in the blood of
chickens of the second group decreased by 18%, the hemoglobin content by 19.5%, and the
number of platelets by 22.1% compared to the blood values of chickens of the second
group. pure control group, and the number of leukocytes from the blood parameters of
chickens of the first group did not matter
The quantity of eosinophils in the leukocyte formula increased by 31.2%,
pseudoeosinophils by 54.2%, monocytes by 43.4%, and the number of lymphocytes
decreased by 15.2% P<0.05. By the end of the experiment, the number of basophils did not
differ from the blood parameters of chickens of the first group P < 0.05.
By the 7th day of the experiment, the number of erythrocytes in the blood of chickens of
the second group decreased by 48.3%, the amount of hemoglobin decreased by 42.3%, but
the number of leukocytes increased by 9.0% and the number of platelets increased. by 8.0%
Eosinophils in the leukocyte formula increased by 53.3% P<0.02, pseudoeosinophils by
66.0% P<0.02, the number of monocytes by 50.0% P<0.05%, and the number of
lymphocytes increased by 31.0 % P<0.02<0.02 decreased.
By the 10th day of the experiment, the number of erythrocytes and the amount of
hemoglobin in the blood of chickens of the 2nd group did not differ from the blood
parameters of chickens of the 1st group. But the number of leukocytes decreased by 42.2%
P<0.01 and platelets by 33.5% P<0.05. The number of eosinophils in the leukocyte formula
did not differ almost significantly from the blood parameters of chickens of the first group.
But pseudoeosinophils increased by 41.0% P<0.05, monocytes by 64.0% P<0.05%, and the
number of lymphocytes decreased by 26.4% P<0.05.
By the 15th day of the experiment, the number of leukocytes in the blood increased by
20.5% P<0.05, and the number of platelets increased by 15.1% compared to the blood
values of the first group of chickens.
Among the types of leukocytes in the leukocyte formula, pseudoeosinophils increased
by 15.0%, monocytes by 17.4% compared to the blood indicators of the first group of
chickens, and the number of lymphocytes decreased by 16.0%.
By the 20th day of the experiment, the morphological blood parameters and the number
of types of leukocytes in the leukocyte formula did not differ from the blood parameters of
chickens of the first comparative pure control group. P<0.05.
Judging by the data obtained as a result of laboratory experiments, with the combined
disease of colibacillosis in chickens, changes in the morphological parameters of the blood
and leukocyte formula were observed in the blood of chickens from the comparative control
group, infected with pathogens and not subjected to treatment. But given that coccidiostats,
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phencid drugs and phencid premixes used in the treatment of diseases do not have a
negative effect on morphological blood parameters and leukocyte formula, these drugs can
be used for the treatment and prevention of diseases in future industrial practice.
4 Discussion
Eimeria pathogens multiply intensively in the mucous membranes of the intestinal system
of chickens, exit with feces and damage the external environment. Drugs used against these
pathogens reduce the intensity of invasion several times.
In our laboratory experiments, it was observed that the intensity of the main invasion
was high in the control group infected with pathogens, that is, in the control group not
exposed to treatment, therefore, on days 5-7 of the experiment, 1 million 320 0000-1
million 820 000 oocysts were isolated from 1 g of litter. When chickens of the experimental
groups were infected with pathogens of Eimeria and colibacillosis and received 288 mg/kg
of the phencidal synergistic mixture and 1000 mg/kg of the phencidal premix with food for
10-12 days, the intensity of the invasion decreased by 4-5 times. The results obtained [6]
are consistent with the information given in [7,8,9,13].
When studying the morphological parameters of blood and the effect of the premix
Phencid on the leukocyte formula, which was used in the experiment to create an
associative course of diseases of eimeria and colibacillosis in chickens, until the end of the
20-day experiment, the morphological parameters of the blood and the numbers obtained
from the leukocyte formula were significantly different from the indicators blood of
chickens from the pure control group. But in chickens of the control group that did not
receive treatment, morphological blood parameters and changes in the leukocyte formula
were observed. Results obtained during the experiment [12] et.others corresponds to the
information
5 Conclusion
Synergistic mixtures of phencid preparations and its premixed form reduce the intensity of
invasion by almost 4-5 times.
Phencid and phencid premix in the treatment of eimeriosis and colibacillosis do not
adversely affect the morphological parameters of the blood of chickens, as well as the
leukocyte blood count.
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