Comparative Anticoccidial Effect of some Natural Products against Eimeria spp. Infection on Performance Traits, Intestinal Lesion and Occyte Number in Broiler

Alaeldein M. Abudabos1,*, Abdullah H. Alyemni2, Elsayeid O. Swilam1 and Mu’ath Q. Al-Ghadi3

1Department of Animal Production, King Saud University, Riyadh, Kingdom of Saudi Arabia

2ARASCO for Feed, P.O. Box 53845, Riyadh 11593, Kingdom of Saudi Arabia

3Zoology Department, King Saud University, Riyadh, Kingdom of Saudi Arabia

ABSTRACT

This study was performed to find the effect of some natural products (Cozante, Norponin and Organimix) in comparison to standard anticoccidial drug (Elancoban) in experimentally induced coccidiosis in broiler. A total 200 birds (Ross 308) were randomly assigned to 10 treatments on day 15 after introduction of coccidiosis. The results indicated that feed intake and production efficiency factor (PEF) did not differ significantly (P>0.05) between Elancoban and natural products. However, treatment had a significant effect on BWG and FCR (P<0.01 and P<0.001, respectively), chicks which had received Elancoban had the highest BWG (327.9 g) and converted the feed to gain with highest efficiency (1.444 g: g). Challenged birds had longer small intestines and heavier liver as compared to unchallenged birds. Lesion score and oocysts secretion increased significantly in the challenged birds, however, Elancoban had no significant difference with the natural products. It was concluded from the present study that natural products could be effective in comparison with anticoccidial drug to control coccidiosis in broiler chicken.

Article Information

Received 16 March 2017

Revised 01 May 2017

Accepted 12 May 2017

Available online 13 October 2017

Authors’ Contribution

AMA, AHA planned and conducted the growth trials. AMA, AHA, EOS and MQA collected samples and did lab analysis.

Key words

Coccidiosis, Broiler, Drugs, Natural products, Control.

DOI: http://dx.doi.org/10.17582/journal.pjz/2017.49.6.1989.1995

* Corresponding author: [email protected]

0030-9923/2017/0006-1989 $ 9.00/0

Copyright 2017 Zoological Society of Pakistan

Introduction

 

Coccidiosis is one of the most dangerous intestinal protozoan diseases in broiler resulting in stunt growth and feed efficiency (Tanweer et al., 2014; Chand et al., 2016). Moreover, the disease makes the bird more vulnerable to other bacterial infections such as Clostridium perfringens. The poultry farmers use largely coccidiostats and vaccine to control the disease. It is believed that the use of coccidiostats has resulted in major breakthrough in poultry industry, however, their wide spread and misuse led to the development of drug resistance against Eimeria. Additionally, strong evidence of the presence of the residues of coccidiostats in the meat of treated birds has not been adequately addressed. Therefore, there is a strong desire to use some natural alternative agents to replace the existing coccidiostats. Although the use of vaccine has shown promising results in poultry industry, however, on a commercial scale, their effect is limited largely due to high production cost and ineffectiveness in the case of poor management conditions.

Preventing a disease is much a better option than fighting it. Natural dietary supplements may potentially be used as one of the novel approaches to treat coccidiosis due to their natural origin, wide dose range, and lack of grace period and stimulation of appetite among others (Chand et al., 2016; Khan et al., 2016; Alzawqari et al., 2016; Raza et al., 2016; Abudabos et al., 2017). A number of plant derived compounds have been successfully used as the natural products to prevent the incidence of coccidiosis (Abudabos et al., 2016a, b). The natural products may act as alternative therapeutic agents against coccidiosis since the new chemical compounds have not been reported to possess. Therefore, the objective of the present work was to compare the effect of natural product with anticoccidial drug and growth performance, lesion score of intestines and occyte shedding in broiler chicks.

 

Materials and methods

 

This experiment was approved by the Departmental board of Studies on Ethics, Methodology and Welfare, King Saud University, Kingdom of Saudi Arabia.

 

Table I.- Dietary ingredients and chemical composition of starter and finisher diets.

	Day 1-14	Day 15-30
Ingredients (%)
Corn	53.21	60.75
Soybean meal	37.92	25.00
Corn gluten meal	2.00	7.10
Corn oil	2.20	2.80
Dicalcium phosphate	2.30	2.05
Limestone	0.83	0.68
Salt	0.45	0.50
VM Mix1	0.50	0.50
DL-Methionine	0.20	0.10
Lysine-HCL	0.22	0.37
Threonine	0.11	0.10
Choline chloride	0.05	0.05
Total	100	100
Chemical composition
ME (kcal/kg)	3000	3150
Crude protein (%)	23.5	21.30
Methionine (%)	0.55	0.44
Lysine (%)	1.42	1.23
Sulfur amino acids (%)	0.96	0.80
Threonine (%)	0.95	0.85
Calcium (%)	1.05	0.90
Phosphorus (%)	0.50	0.45

1Vitamin-mineral premix contains in the following per kg: vitamin A, 2400000 IU; vitamin D, 1000000 IU; vitamin E, 16000 IU; vitamin K, 800 mg; vitamin B1, 600 mg; vitamin B2, 1600 mg; vitamin B6, 1000 mg; vitamin B12, 6 mg; niacin, 8000 mg; folic acid, 400 mg; pantothenic acid, 3000 mg; biotin 40 mg; antioxidant, 3000 mg; cobalt, 80 mg; copper, 2000 mg; iodine, 400; iron, 1200 mg; manganese, 18000 mg; selenium, 60 mg, and zinc, 14000 mg.

 

During the starter period, a total of 200, day old broiler chicks (Ross 308) were allotted to 40 cages. A typical isocaloric and isonitrogenous starter (0 to14 d) and finisher (15 to 35 d) diets were formulated in mashed form based on Ross 308 recommendation guide (Table I). Birds received one of the following six treatments: At day 15 of age, half birds received coccidial challenge while the other half had the treatment without challenge which yielded 10 treatments as follow: 1) Negative control, 2) Coccidial challenge (positive control), 3) Elancoban (0.5 kg/ton monensin sodium), without coccidiosis challenge, 4) With Elancoban (0.5 kg/ton monensin sodium), with coccidiosis challenge (positive Elancoban), 5) With Cozante (0.5 kg/ton), without coccidiosis challenge (negative Cozante). Cozante is a polyphenol compound which is highly effective against Eimeria species, 6) Cozante (0.5 kg/ton) with coccidiosis challenge (positive Cozante), 7) Norponin XO (0.5 kg/ton), without coccidiosis challenge (negative Norponin XO). Norponin is a natural herbal extract rich in saponin e.g., protodioscine, schidigera saponin B1, 8) Norponin XO (0.5 kg/ton) with coccidiosis challenge (positive Norponin XO), 9) Organicox (0.5 kg/ton) without coccidiosis challenge (negative Organicox). Organicox is a blend of organic acid and 10) Organicox (0.5 kg/ton) with coccidiosis challenge (positive Organicox).

The challenge was achieved on day 15 by a mixture of sporulated oocysts of pathogenic strains of Eimeria by live oocysts Coccivac®-D (contains live oocysts of Eimeria acervulina, Eimeria mivati, Eimeria maxima, Eimeria tenella, Eimeria necatrix, Eimeria praecox, Eimeria brunette and Eimeria hagani.

Performance measurements

Average body weight gain (BWG) and feed intake (FI) for each pen was recorded every 5 days and average feed conversion efficiency (FCR) were adjusted for mortality and computed. Production Efficiency Factor (PEF) was calculated weekly by using the formula:

 

 

Small intestine and carcass measurements

At day 25 eight birds per treatment were selected for small intestinal measurements, the entire gastrointestinal tract was removed aseptically after slaughtering and defeathering. Small intestine and ceca were weighed and the total length was measured then the small intestine was separated into duodenum, jejunum (proximal to Meckel’s diverticulum) and ileum (proximal to ileo-caecal junction) and for each part measurements of length and weight. Intestine relative weight was calculated as a ratio between total intestinal weight (g) and dressed weight (g).

Coccidial lesion

Coccidial lesion scoring in the ileum, jejunal and cecal regions of the intestine was carried out using the method of Conway et al. (1999). Lesion scores were recorded as 0, 1, 2, or 3, from least to most severe.

Oocyst counting

The oocysts number per gram of excreta (OPG) was determined in pooled excreta samples collected on days 25 post infections. A modified McMaster counting chamber technique was used. Excreta suspension in a salt solution was prepared (10% w/v). Then 1 mL of the suspension was mixed with 9 mL of a salt solution. McMaster chamber was used to count the number of oocysts.

Statistical analysis

The collected data were evaluated with ANOVA for a complete randomized block design, using the general linear models procedure of SAS software (SAS, 2003). The data were analyzed using the General Linear Model procedure of the Statistical Analysis System (SAS, 2003) as a 5 × 2 factorial arrangement of treatments that included 5 dietary treatments (5 levels) and challenge (2 levels), and their respective interactions. LSD test was applied for mean to compare the treatment means when the treatment effect was significant at P <0.05.

 

Results

 

Table II shows that the challenge had a significant impact on all performance parameters. Unchallenged chicks consumed more feed, gained more weight, converted feed more efficiently and as a result had higher FEF as compared to challenged chicks. Chicks received Elancoban had the highest BWG (327.9 g) and converted the feed to gain with highest efficiency (1.444 g: g). Chicks on the control diet were intermediate in terms of BWG and FCR but they were better than Cozante, Norponin and Organimix.

The morphometric measurements of the intestinal samples at 25 d of age (10 days post inoculation) are given in Table III. No significant differences in intestinal length, weight or (intestine relative weight) IRW were observed because of treatment or the interaction between treatment and challenge (P>0.05). However, challenge affected the total length of the small intestine (P<0.01). Challenged birds had longer small intestines as compared to unchallenged birds. Also challenge affected the percentage of cecal length (P<0.05), unchallenged birds had higher cecal length percentage as compared to challenged birds. Challenged birds had higher IRW as compared to unchallenged birds. Liver percentage was affected by treatment and challenge (P<0.05; P<0.001, respectively). Birds received treatment 2 (Elancoban) had lower liver percentage as compared to those which had received control, Norponin or Organicox (P<0.05) but similar to those received Cozante. On the other hand, challenged birds had higher liver percentage as compared to unchallenged birds.

 

Table II.- Live weight (BW), feed intake (FI), feed conversion ratio (FCR) and production efficiency factor (PEF) of broiler chickens given experimental diets at 25 days.

Treatment	TRT	Challenge	Performance
			FI (g)	BWG (g)	FCR (g: g)	PEF
1	Control	No	524.2a	374.5a	1.339b	298.9
2	Control	Yes	423.4d	243.9c	1.740a	255.1
3	Elancoban	No	460.9bcd	323.2b	1.428b	291.9
4	Elancoban	Yes	485.4ab	332.8b	1.459b	302.5
5	Cozante	No	479.7abc	333.9b	1.440b	277.6
6	Cozante	Yes	440.5bcd	251.5c	1.764a	231.4
7	Norponin	No	479.0abc	336.1b	1.426b	297.6
8	Norponin	Yes	436.4cd	247.9c	1.761a	280.1
9	Organicox	No	481.9abc	331.8b	1.451b	291.2
10	Organicox	Yes	420.9d	241.7c	1.743a	269.3
SEM±			17.31	11.98	0.037	14.70
TRT average
Control			473.8	309.3ab	1.569a	276.9
Elancoban			473.1	327.9a	1.444b	297.2
Cozante			460.1	292.7bc	1.602a	254.5
Norponin			457.7	292.0bc	1.593a	288.8
Organicox			451.4	286.8c	1.598a	280.3
SEM±			12.24	8.47	0.027	10.40
Challenge average
No			485.1a	339.9a	1.429b	291.4a
Yes			441.3b	263.6b	1.693a	267.7b
SEM±			7.34	5.08	0.016	6.24
Statistical probabilities
TRT			NS	**	***	NS
Challenge			***	***	***	**
TRT x Challenge			**	***	***	NS

*, p<0.05, **p<0.01; ***, p<0.001; NS, not significant; SEM, Standard error of the mean.

 

Table III.- Effect of different treatments and challenge on small intestinal measurements of broiler chickens at 25 d (10 d post inoculation).

 

Table IV.- Effect of experimental treatments and challenge on lesion score (0-4) at 25 d challenged by coccidia (10 day post inoculation).

Treatment	Trt	Challenge	Duodenum	Jejunum	Ceca
			(points)	(points)	(points)
1	Control	No	0.00	0.00	0.00
2	Control	Yes	3.00	2.33	3.00
3	Elancoban	No	0.00	0.00	0.00
4	Elancoban	Yes	1.67	1.33	1.00
5	Cozante	No	0.00	0.33	0.00
6	Cozante	Yes	2.00	1.33	2.33
7	Norponin	No	0.00	0.33	0.00
8	Norponin	Yes	2.33	2.33	2.67
9	Organimix	No	0.00	0.33	0.00
10	Organimix	Yes	2.67	3.00	3.00
SEM±			0.316	0.333	0.350
Trt. Average
Control			1.50	1.17	1.50
Elancoban			0.83	0.67	0.50
Cozante			1.00	0.83	1.17
Norponin			1.17	1.33	1.33
Organimix			1.33	1.67	1.50
SEM±			0.224	0.236	0.247
Challenge average
No			0.00b	0.20b	0.00b
Yes			2.33a	2.07a	2.40a
SEM±			0.141	0.149	0.156
Statistical probabilities
Trt			NS	NS	NS
Challenge			***	***	***
Trt x Challenge			NS	NS	NS

*, p<0.05; **, p<0.01; ***, p<0.001, NS, not significant; SEM, standard error of the mean.

 

Table IV shows the result of lesion scores in duodenum, jejunum and ceca of birds at 25 of age. Generally, uninfected birds (control, unchallenged) were free of lesions. At day 25, coccidial challenge affected lesion score at duodenum jejunum and ceca (P<0.001). Lesions were higher in challenged group as compared to unchallenged group for all sections measured.

Table V indicates the effects of different feed additives and challenge on the oocyte output. The results indicated that the natural feed additives performed better than and no significant difference was noticed between Elancoban and other feed additives.

 

Discussion

 

Search for the alternatives to anticoccidial drugs to control coccidiosis is an important field of study in poultry production. Intestinal integrity of broiler may be compromised during exposure to pathogens of coccidiosis. This disease impacts both the performance and health of the birds leading to significant economic losses.

This study was designed to find the success of feed additives from natural sources in comparison with a standard anticoccidial drug against Eimeria spp. As expected, the performance was most negatively affected by the Eimeria challenge in the positive control group. Dietary supplementation of natural additives improved the feed efficiency and weight gain almost similar to the birds treated with anticoccidial drug. The efficient role of anticoccidials derived from plants for the controlling of coccidiosis has been previously documented (Abbas et al., 2012; Bozkurt et al., 2014). The positive effects of the use of botanicals on the performance has been linked to the active compounds which alter miroflora modulation, reduce oocysts shedding, decrease intestinal inflammation, enhance immunity and improve antioxidant status (Chand et al., 2014; Tehseen et al., 2016). The effectiveness of herbal additives alleviates the destructive effect of coccidial infection and maintains growth. Confirming our report, previous studies have also documented improved performance in birds in response to anticoccidials (Garcia and Bolis, 2005; Küçükyilmaz et al., 2012). In the current study, the increased liver and intestine relative weight increased significantly in the infected birds showing the damaging effect of the infection.

 

Table V.- Effect of different treatments and challenge on oocyst output of broilers (10 days post-inoculation).

Treatment	Trt	Challenge	PI-10 (Log10 per g excreta)
1	Control	No	0.00
2	Control	Yes	3.71
3	Elancoban	No	0.00
4	Elancoban	Yes	2.92
5	Cozante	No	0.00
6	Cozante	Yes	3.05
7	Norponin	No	0.00
8	Norponin	Yes	2.97
9	Organimix	No	0.00
10	Organimix	Yes	3.18
SEM±			0.104
Trt. Average
Control			1.86a
Elancoban			1.46b
Cozante			1.53b
Norponin			1.48b
Organimix			1.59b
SEM±			0.073
Challenge average
No			0.00b
Yes			3.17a
SEM±			0.046
Statistical probabilities
Trt			**
Challenge			***
Trt*Challenge			**

*, p<0.05; **, p<0.01; ***, p<0.001, NS, not significant; SEM, standard error of the mean.

 

In the present study, the lesion score was significantly reduced in the duodenum, jejunum and caeca of the infected birds in response to treatments showing the effectiveness of the natural additives. The positive effect of herbal mixture may be due to the presence of flavonoids, tannins, alkaloids and saponins, which exhibit anti-inflammatory, antioxidant and anti-parasitic properties (Khan et al., 2012; Tanweer et al., 2014).

In the current study, the oocysts shedding after 10 days of the infection were similar in the anticoccidial drug and natural products. Confirming our work, several earlier studies have reported efficient reduction in the fecal oocysts in birds challenged with experimental coccidial infection in response to natural products (Christaki et al., 2004; Ibrir et al., 2009; Bozkurt et al., 2012; Almeida et al., 2014; Tanweer et al., 2014).

It was concluded from the present study natural products could be effective in comparison with anticoccidial drug to control coccidiosis in broiler chicken.

 

Acknowledgement

 

The authors would like to extend their sincere appreciation to the Deanship of Scientific Research at King Saud University for its funding of this research through the Research Group Project No. RGP-273.

 

Statement of conflict of interest

Authors have declared no conflict of interest.

 

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