COMPARISON OF THE EFFECTS OF HERBAL COMPOUNDS AND CHEMICAL DRUGS FOR CONTROL OF COCCIDIOSIS IN BROILER CHICKENS
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Bulgarian Journal of Veterinary Medicine, 2020 ONLINE FIRST
ISSN 1311-1477; DOI: 10.15547/bjvm.2020-0005
Original article
COMPARISON OF THE EFFECTS OF HERBAL COMPOUNDS
AND CHEMICAL DRUGS FOR CONTROL OF COCCIDIOSIS
IN BROILER CHICKENS
S. S. MOUSAVINASAB1, M. H. BOZORGMEHRIFARD2,
S. M. M. KIAEI2, S. RAHBARI3 & S. CHARKHKAR2
1
DVSc Resident in Hygiene and Diseases of Poultry; Department of Clinical
Sciences; 2Department of Clinical Sciences; 3Department of Pathobiology,
Faculty of Veterinary Medicine, Science and Research Branch, Islamic Azad
University, Tehran, Iran
Summary
Mousavinasab, S. S., M. H. Bozorgmehrifard, S. M. M. Kiaei, S. Rahbari & S. Charkhkar,
2020. Comparison of the effects of herbal compounds and chemical drugs for control of
coccidiosis in broiler chickens. Bulg. J. Vet. Med. (online first).
Coccidiosis is the most important intestinal parasitic disease of broiler chickens in poultry industry.
Because of the increasing resistance to anticoccidial agents and presence of their residues in meat and
eggs, it is necessary to find safe and new anticoccidial compounds. This study was conducted to com-
pare the effects of two herbal compounds, including Artemisia sieberi and Curcuma longa, and their
mix with a chemical anticoccidial drug on broilers’ performance during a mixed coccidian challenge.
A total of 216, one-day-old Ross 308 broilers were randomly divided into six groups. Different herbal
extracts and one chemical anticoccidial agent were used in each group. Five groups were infected
with a mixture of Eimeria sporulated oocysts at the age of 21 days with crop gavage. Body weight
and feed intake were measured then feed conversion ratio was calculated on a weekly basis. Mortality
was recorded when occurred throughout the experimental period. Oocysts excretions and lesion
scores were investigated weekly up to three weeks after infection. Eimeria-challenged birds had a
reduction in growth parameters compared to the uninfected birds (PComparison of the effects of herbal compounds and chemical drugs for control of coccidiosis in …
et al., 2010; Gilbert et al., 2011; Pop et products have proved to be less toxic and
al., 2019). Due to Eimeria multiplication thought to be ideal feed additives in food
in the intestinal tract, the parasite causes animal production (Puvača et al., 2018).
tissue damage. This can disturb the feed- Because of an increasing demand for
ing, digestive processes and nutrient ab- natural herbal food products, a lot of
sorption, leading to signs of dehydration, herbal extracts are studied for the treat-
blood loss, poor skin pigmentation and ment of poultry coccidiosis to find new
increased susceptibility to other diseases. alternatives of the traditional anticoccidial
Clinical signs include diarrhoea or soft, drugs (Masood et al., 2013)
mucoid faeces, poor growth, impaired Artemisinin and its derivatives can be
feed conversion ratio and increased mor- antimalarial agents. Artemisinin was iso-
tality (Hafez, 2008). lated for the first time in China in 1972
Eimeria has a varying pathogenicity, from the leaves of Artemisia annua. Seve-
with some invading deep in the intestinal ral studies indicated that it had impressive
mucosa, causing wide-spread damage and activities, with high efficacy against multi-
distinct gross lesions and others being less drug-resistant forms of malaria parasites
destructive but still having a significant with negligible toxicity and side-effects
effect on productive performance (Morris (White, 1994). Artemisinin (Wiedosari &
& Gasser, 2006). To prevent coccidiosis Wardhana, 2017) and Artemisia sieberi
and minimise related economic losses, extract (Arab et al., 2006; Cheeke, 2009)
anticoccidial drugs such as polyether have anticoccidial effects. The ability of
ionophores, sulfanamides and chemical artemisinin to bind to membrane choles-
drugs are applied via feed prophylacti- terol of protozoan cells allows modifying
cally (De Gussem, 2007). Due to their membrane function and structure and ac-
regular use, Eimeria strains have become count for their antiprotozoal activity
drug-resistant (Peek & Landman, 2003); (McAllister et al., 2001).
Moreover, there is strong evidence that Curcuma longa, commonly known as
residues of some coccidiostats may be turmeric, is a medicinal plant widely cul-
present in the meat and egg and so the tivated and used in the tropical regions.
consumer is not adequately protected Traditionally, it has been used to treat
(Olejnik et al., 2009). Therefore, alterna- various diseases (Abbas et al., 2012).
tive strategies for coccidiosis control are Plant extracts have been found to have
needed besides the application of coccidi- antioxidative (Puvača et al., 2018) and
osis vaccines that are expensive. In this imunomodulatory effects (Antony et al.,
regard, herbal compounds seem to have 1999). The antioxidant effect of a herbal
some potential in controlling coccidiosis product is directly linked with its anticoc-
(Abbas et al., 2012). Herbal remedies cidial effect (Alhotan & Abudabos, 2019).
have been used in medicine for as long as Curcumin (diferuloylmethane) is a natural
human history. They have recently gained polyphenolic compound abundant in the
increasing popularity especially because rhizome of the perennial herb turmeric
of the declining effectiveness of synthetic that is found in Curcuma longa roots in
compounds in addition to concerns of concentrations ranging from 1% to 5%
consumers about drug effects (Kim et al., (Conney et al., 1991). Curcumin is known
2013). Compared with synthetic antibio- to augment antioxidant status especially
tics or inorganic chemicals, plant-derived through superoxide dismutase (SOD)
2 BJVM, ××, No ×S. S. Mousavinasab, M. H. Bozorgmehrifard, S. M. M. Kiaei, S. Rahbari & S. Charkhkar
which could be due to the increased ex- with internationally recognised guidelines
pression of SOD gene in the chickens fed for animal welfare and the principles and
turmeric. Antioxidant enzymes, such as specific guidelines presented in the Guide
catalase (CAT) within the peroxisomes for the Care and Use of Laboratory Ani-
and cytosolic glutathione peroxidase mals (Anonymous, 2011). A total of 216
(GPx), are involved in the conversion of one-day-old Ross 308 broiler chicks after
hydrogen peroxide, a powerful and poten- weighing were divided into six experi-
tially harmful oxidising agent into water mental groups with three replicates for
and molecular oxygen (Kostadinović et each group (12 chicks in each replicate).
al., 2015; Puvača et al., 2018). Abbas et The chicks were reared in a building with
al. (2010) also had reported the anticoc- floor pens on with built-up wood shavings
cidial activity of Curcuma longa. as bedding and whole house lighting. The
Kim et al. (2013) evaluated the effects lighting programme during the experimen-
of dietary supplementation with an or- tal period was 23L:1D. The groups that
ganic extract of Curcuma longa in com- underwent the Eimeria challenge received
mercial broiler chickens. They saw that different treatments beginning from five
this dietary supplementation enhanced days before challenge until five days be-
coccidiosis resistance as demonstrated by fore being slaughtered.
increased body weight gain, reduced fae- The experimental groups were as fol-
cal oocyst shedding (OPG) and decreased lowed: Group 1: Uninfected untreated
gut lesions. According to Abbas et al. control group (negative control); Group 2:
(2012) in Pakistan, small broiler farmers Infected untreated control group (positive
add Curcuma longa powder to the feed to control); Group 3: Amprolium 25% +
control coccidiosis in broilers. ethopabate 1.6% (Ethoamprox® Rooyan
Since the most active ingredient of Darou Pharmaceutical Co., Tehran, Iran)
both Artemisia sieberi and Curcuma 125 + 8 ppm in feed; Group 4: Ethanol
longa extracts (artemisinin and curcumin, extract of Artemisia sieberi 5 mL/L in
respectively) is responsible for their bio- drinking water (1.1 mg/mL artemisinin);
logical activity and its anticoccidial ef- Group 5: Ethanol extract of and Curcuma
fects, we hypothesised that their combina- longa 5 ml/L in drinking water (3.95
tion could have better suppressive effects mg/mL curcumin); Group 6: Ethanol ex-
and act against mix infections of Eimeria tract of Artemisia Sieberi and Curcuma
oocysts in broiler chickens. This study longa 10 mL/L in drinking water (5 mL/L
was conducted to evaluate and compare Artemisia sieberi extract and 5 mL/L Cur-
the prophylactic and controlling effects of cuma longa extract). The herbal extracts
Artemisia sieberi (artemisinin), Curcuma of Artemisia sieberi and Curcuma longa
longa (curcumin) and their combination were prepared by the same extraction pro-
with amprolium ethopabate to treat avian cedure for both plants (Almeida et al.,
coccidiosis in broiler chickens. 2014). The content of active ingredients
was estimated by HPLCUV according to
Ferreira & Gonzales (2009).
MATERIALS AND METHODS
Table 1 describes the feed and nutrien-
This study was conducted in the Poultry tional composition of all experimental
Research Center, Faculty of Agriculture of diets. A three-phase feeding programme
Tarbiat Modares University in accordance was applied, with starter, grower and fin-
BJVM, ××, No × 3Comparison of the effects of herbal compounds and chemical drugs for control of coccidiosis in …
Table 1. Composition of the diets (as-fed basis): starter (day 110), grower (day 1128), and finisher
(day 2942)
Item Starter Grower Finisher
Ingredients (%)
Corn 58.62 62.84 65.74
Soybean meal (44%) 35.55 32 29
Soybean oil 1.2 1.5 1.8
Dicalcium phosphate 1 1.2 1.1 1.1
CaCO3 (38%) 1.3 1.2 1.1
Sodium chloride 0.3 0.3 0.3
L-Lysine HCl 0.2 0.14 0.15
DL-methionine 0.27 0.2 0.19
L-Treonine 0.11
Choline chloride 0.7 0.17 0.083
Phytase 0.05 0.05 0.05
Vitamin premix 2 0.25 0.25 0.25
Mineral premix 3 0.25 0.25 0.25
Contents by calculation
ME (kcal/kg) 2940 3025 3080
CP (%) 21.45 20.15 19.08
Lys (%) 1.39 1.15 1.11
Met (%) 0.98 0.89 0.83
Met + Cys (%) 1.03 0.91 0.85
Available phosphorus (%) 0.49 0.46 0.44
Calcium (%) 0.98 0.89 0.88
1
Contains 20% P and 23% Ca; 2Vitamin premix provided the following (per kg of diet): 12,000 IU of
retinyl acetate, 5,000 IU of cholecalciferol, 80 IU of dl-α-tocopheryl acetate, 3.2 mg of menadione
sodium bisulfite, 3.2 mg of thiamine, 8.6 mg of riboflavin, 60 mg of nicotinic acid, 17 mg of calcium
d-pantothenate, 5.4 mg of pyridoxine, 2.2 mg of folic acid, 0.02 mg of cyanocobalamine; 3Trace
mineral premix provides the following (per kg of diet): 250 mg of choline chloride, 0.17 mg of bio-
tin, 120 mg of MnSO4.H2O, 20 mg of FeSO4.7H2O, 110 mg of ZnO, 16 mg of CuSO4.5H2O, 1.25 mg
of iodised NaCl, 0.3 mg of Na2SeO3.
isher (from 110, 1128, and 2942 days use. Coccidial oocysts were enumerated
respectively). All groups had the same before challenge. On the 21st day of age,
corn-soybean meal basal diet. Feed and each bird in groups 2 to 6 was challenged
water were offered ad libitum. by inoculating with 0.5 mL solution of a
The Eimeria oocysts used for chal- mixture suspension of fresh sporulated
lenge in the present study were from field oocysts of pathogenic strains of Eimeria
Eimeria strains. They were isolated from (containing 4×104 sporulated oocysts of E.
commercial broiler and breeder produc- acervulina, 3×104 sporulated oocysts of
tion houses around Tehran. The oocysts E. maxima and 5×104 sporulated oocysts
were preserved in 2.5% potassium di- of E. tenella per bird). The negative con-
chromate solution to induce sporulation trol group was inoculated with 0.5 mL of
o
and kept in a refrigerator (25 C) until normal saline solution.
4 BJVM, ××, No ×S. S. Mousavinasab, M. H. Bozorgmehrifard, S. M. M. Kiaei, S. Rahbari & S. Charkhkar
Efficacies of A. sieberi, C. longa and For the statistical analysis, Shapiro-
amprolium ethopabate to ameliorate dele- Wilk and Bartlett tests were used to
terious effects of coccidial infection were evaluate similarity and normality of the
evaluated on the basis of body weight gain data. Analysis of variance (ANOVA) was
(BWG), feed intake (FI), mortality rate used to compare among the study groups.
(MR), feed conversion ratio (FCR), Euro- Bonferroni test was used for paired data.
pean efficiency factor (EEF), oocyst ex- In case the data were not distributed nor-
cretion (oocyst per gram of faeces = OPG) mally, a non-parametric test like Kruskal-
and lesion scoring. Body weight gain and Wallis was used. The significance level
feed intake of the chickens in each group was considered less than 0.05 in all tests.
and their replicates were determined
weekly up to the end of experiment (six
RESULTS
weeks). Mortality and culls were recorded
for the entire study period (days 042). The results of broiler performance pa-
Daily mortality record was maintained and rameters of the current study are presented
the birds were not replaced. Body weight in Table 2. During the first 21 days of
gain and feed intake were also calculated experiment, before inoculation of infec-
before and after the challenge. The Euro- tion, the body weight gain, feed intake and
pean efficiency factor was calculated at feed conversion ratio were not signifi-
the end of experiment period by the fol- cantly different among the groups. In the
lowing formula: EEF = 100×[Viability post-challenge period (29 to 42 days), all
(%) × body weight (kg)] / [age, days × groups except the negative control group
FCR (kg feed/kg gain)]. had a reduced weight gain and feed intake
At 7th, 14th and 21st days after the chal- as well as increased feed conversion ratio.
lenge, samples of faeces were collected by Significant differences were observed
placing a white card in each pen. Faecal between negative control and other ex-
samples were diluted in sugar solution perimental groups (P6
Table 2. Effect of anticoccidial drug and herbal extracts on the performance of coccidia-challenged broilers (mean±SD; n=36)
Treatment
P-
Item Period
Negative Positive Amprolium A. sieberi + value
A. sieberi C. longa
control control ethopabate C. longa
a
BW (g) 121 day 839.4 ± 21.2a 827.1 ± 7.3a 839 ± 10.5a 836.6 ± 5 837 ± 8.9a 837.6 ± 4a 0.766
2142 day 1625.7 ±4.7a 1145.5 ± 37.8d 1529.5±48.5ab 1258 ± 4cd 1280.6 ±31.2c 1431.3 ± 18.9bS. S. Mousavinasab, M. H. Bozorgmehrifard, S. M. M. Kiaei, S. Rahbari & S. Charkhkar None of the herbal extracts was able to Fig. 1. On day 28, oocyst excretion count achieve a similar result to amprolium of amprolium ethopabate group was sig- ethopabate group. BW and FI in groups nificantly lower than the positive control which received herbal extracts were sig- group and relatively lower than the other nificantly lower vs the amprolium ethpa- groups (P
Comparison of the effects of herbal compounds and chemical drugs for control of coccidiosis in … The group that received C. longa extract (2006; 2012) reported that Artemisia had higher lesion scores in the caecal part sieberi and granulated A. sieberi extract of intestine, but the effect of this extract decreased oocyst shedding (OPG) and on lesions of the middle part of intestine severity of coccidial infection. Likewise, was better compared to the A. sieberi Allen et al. (1997) reported that A. annua group. The lesion scores of the mixed reduced the intensity of E. tenella and E. herbal extract group were relatively better acervulina infections and strongly de- than the other herbal groups, but not better creased oocyst excretion per gram. than the amprolium ethpabate group Almeida et al. (2014) also found a signifi- (P
S. S. Mousavinasab, M. H. Bozorgmehrifard, S. M. M. Kiaei, S. Rahbari & S. Charkhkar
et al (1997, 1998), Arab et al. (2006) and results were similar to the results of this
Wiedosari & Wardhana (2017). Kheira- study in terms of oocyst excretion and
badi et al. (2014) also reported improved broilers’ performance.
growth performance parameters and sig- On the 5th day after challenge, the le-
nificantly decreasing oocyst excretion in sion scores in herbal groups were rela-
Coccidia challenged broilers fed granu- tively higher than the anticoccidial drug
lated extract of A. sieberi compared to an group. Yet, on 35th and 42nd days of age,
anticoccidial drug. the scores of herbal groups were close to
Curcuma longa, the other extract plant those of the chemical anticoccidial group,
which was used in this study has anti- showing that the herbal compounds had
protozoan properties (Shahiduzzaman et delayed effects compared to anticoccidial
al., 2009). Curcumin is a natural polyphe- drugs. Lower lesion scores were observed
nolic compound with substantial antioxi- with A. sieberi and C. longa extracts mix,
dative effects. Alhotan & Abudabos e.g. these extracts had a synergistic effect
(2019) showed that the antioxidant capa- in improving intestinal damage of due to
city of the herbal product is directly linked infection with Eimeria oocysts.
to its anticoccidial effect. By comparing It could be concluded that although the
the lesion scores of A. sieberi and C. effectiveness of using Artemisia sieberi
longa groups, it was realised that C. longa and Curcuma longa extracts alone or to-
extract had a better effect on treating the gether was inferior to that of Amprolium
lesions of the middle part of intestine ethopabate for decreasing deleterious ef-
compared to A. sieberi. Allen et al. (1998) fects of coccidiosis in broilers and im-
and Kim et al. (2013) reported a reduction proving the productive performance, these
in lesion scores and oocyst excretion by herbal compounds, and especially their
using 1% C. longa in diets of broilers mixture could be effective. Finally, further
challenged with E. maxima oocysts and studies on the mix of other anticoccidial
enhanced coccidiosis resistance as evi- herbal compounds are recommended.
denced by increased body weight gains. In
agreement with the results of the present
study, Khalafalla et al. (2011) reported ACKNOWLEDGEMENTS
that curcumin could reduce E. tenella’s
The authors wish to thank Seyed Nouraddin
destructive effects and it had a marked
Mousavinasab for doing the statistical analysis
inhibitory in vitro effect on E. tenella and Reza Taherkhani for feed technical sup-
sporozoites, inducing morphological port during the study. Also, thanks for the
changes and reducing sporozoite viability technical assistance of the staff of the parasi-
and infectivity. The same happened re- tology laboratory of the Faculty of Veterinary
garding oocyst excretion per gram. Abbas Medicine, Science and Research Branch, Is-
et al. (2010) reported that by challenging lamic Azad University, Tehran. They ac-
broilers with Eimeria and using 3% C. knowledge the University of Tarbiat Modares
longa powder supplementation in the for giving place to do this research. Last but
not the least, they also thank Muhammed Hus-
feed, milder bloody diarrhoea occurred
sein Mousavinasab for editing this work.
and the body weight gain and feed con-
sumption were significantly higher, also
oocyst excretion counts is significantly
lower than positive control group. Their
BJVM, ××, No × 9Comparison of the effects of herbal compounds and chemical drugs for control of coccidiosis in …
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