Effect of Aqueous Extract of Cichorium intybus as Alternative Antibiotic Growth Promotor on Growth Performance, Lipid and Mineral Profiles of Broilers

Muhammad Yousaf1,2, Muhammad Sarwar1, Irfan Ahmed2,3*, Zahid Kamran2, Dalia Fouad4, Shahzad Ashraf1, Farid Shokry Ataya5, Shakeel Ahmad6,

Abdur Rahman7, Mubarik Mahmood7, Ahmad Kamran Khan8 and

Hafiz Ishfaq Ahmad9

1Institute of Animal Nutrition and Feed Technology, University of Agriculture, Faisalabad, Pakistan

2Department of Animal Nutrition, Faculty of Veterinary and Animal Sciences, The Islamia University of Bahawalpur, Bahawalpur, Pakistan

3Yunnan Provincial Key Laboratory of Animal Nutrition and Feed, Yunnan Agricultural University, Kunming 650201, Yunnan Province, People’s Republic of China

4Department of Zoology, College of Science, King Saud University, PO Box 22452, Riyadh 11495, Saudi Arabia

5Department of Biochemistry, College of Science, King Saud University, PO Box 2455, Riyadh 11451, Saudi Arabia

6Department of Poultry Science, Faculty of Veterinary and Animal Sciences, The Islamia University of Bahawalpur, Bahawalpur, Pakistan

7Department of Animal Sciences, CVAS-Jhang 35200, University of Veterinary and Animal Sciences, Lahore 54000, Pakistan

8Department of Plant Protection, Ghazi University, Dera Ghazi Khan, Punjab, Pakistan

9Department of Animal Breeding and Genetics, Faculty of Veterinary and Animal Sciences, Bahawalpur, Pakistan

ABSTRACT

This study aimed to determine the beneficial effect of chicory leaves extract on growth performance, lipids and minerals profiles of broilers at different pH levels. The influence of Cichorium intybus (chicory) leaf extract was determined on day-old chicks (n=150) that were divided into 15 groups, each of 10, for 35 days. These replicates were allotted to five treatment groups, each receiving three replicates. Two experimental diets (starter and finisher) were formulated with and without antibiotic growth promoter (AGP) Enradin and coccidiostat Salinomycin. Birds of positive control (PC) were offered commercial ration supplemented with AGP and Salinomycin along with water without any supplementation, while birds of negative control were fed ration without AGPs but given fresh water without any extract. Moreover, birds of acidic chicory leave extract, neutral chicory leave extract and basic chicory leave extract (BCLE) groups were fed diets without AGPs but given water supplemented with chicory leave aqueous extract, at pH 3, 7 and 12, respectively. Supplementation of chicory leaf extract significantly influenced (P<0.05) the weight gain and FCR in BCLE economically better than controls. However, feed intake, serum minerals (Ca, P, Na, and K), and mortality were found non-significantly (P>0.05) affected by the chicory leaf aqueous extract. Replacement of AGPs with chicory leaves aqueous extract, extracted at different pH, revealed better performance of broilers in terms of weight gain and efficiency of feed utilization along with better profit margin. Therefore, the use of chicory extract in broiler production may be recommended as an inexpensive but efficient alternative to AGPs.

Article Information

Received 12 September 2023

Revised 15 November 2023

Accepted 04 December 2023

Available online 28 February 2024

(early access)

Published 06 May 2025

Authors’ Contribution

Data curation was made by MY, IA AKK, MM, AR. Formal analysis was made by ZK, SA, HIA, SA. Methodology was made by MY, MS, IA, FSA. Software was assigned to ZK, AKK, SA, AR, MM. Writing original draft was done by MY, IA, DF, HIA. Writing review and editing was done by MY, IA.

Key words

Antibiotic growth promoter, Antibiotics, Chicory, Growth performance, Lipid profile, Broiler

DOI: https://dx.doi.org/10.17582/journal.pjz/20230912065722

* Corresponding author: [email protected]

0030-9923/2025/0003-1279 $ 9.00/00

Copyright 2025 by the authors. Licensee Zoological Society of Pakistan.

This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

Introduction

Recent studies indicated that because of extensive use of antibiotics as growth promoters, there is a risk of developing cross-resistance and multiple-antibiotic resistance in harmful bacteria and many other harmful effects (Verdonk et al., 2005; Mahdi et al., 2022). Antibiotics like tetracycline are the major cause of liver injury which not only alter liver functions but also disturb the metabolism of primary nutrients (Andrade and Tulkens, 2011). It has been noticed that blood triglyceride concentration may be altered because of an injured or affected liver. Antibiotics have also been reported to disrupt the gut microflora and may allow growth of pathogens leading to compromised production (Levy, 2000). Reduced absorption of minerals has also been reported in birds receiving antibiotics as a growth promoter, which might be attributed to changes in gut environment because of antibiotics failure (Klevay and Milne, 2002). Due to these adverse effects of antibiotics on health and its transmission to the human food chain, the European Commission has phased out and finally banned the selling and use of antibiotics as growth promoters in feed (EC Regulation No. 1831/2003). Poultry nutritionists now-a-days are looking for new substitutes to antibiotic growth promoters that have minimum side effects for poultry which could promote growth and can be equally effective against pathogenic microorganisms of gastrointestinal tract by enhancing weight gain and feed efficiency (Ahmed et al., 2022). Due to ban on the use of antibiotics, there is a shift in focus and the use of biological products, such as prebiotics, probiotics, enzymes, organic acids, synbiotics and plant extracts (phytobiotics), as alternatives to antibiotic feed additives in diets for non-ruminant or monogastric animals, has found place in poultry feed (Bedford, 2000). Amongst the various alternatives, plant aqueous extracts are valuable to reducing these problems. Some plants have natural effects like tonics, stimulants, carminative, antiparasitic, antimicrobial, antibacterial, antifungal, and antiseptic (Ferdous et al., 2019).

Cichorium intybus (Chicory) is an herbal medicinal plant of the Asteraceae family (Ivarsson et al., 2011), containing inulin, flavonoids, coumarins, fructo-oligosaccharides and many vitamins that have anti-bacterial, anti-inflammatory, anti-hypercholesterolemia, hepatoprotective, anti-oxidant, anti-fungal, digestive, diuretic, immuno-stimulant, anti-cancer, gastroprotective, and laxative properties without any significant harmful effects (Kalantari and Rastmanesh, 2009). C. intybus is a health-promoting feed ingredient that benefits the host by selectively stimulating the growth and or activity of one or many naturally present or introduced bacterial species in the intestine that can regulate appetite and lipid-glucose metabolism (Glibowski and Skrzypczak, 2017; Song et al., 2018). Chicory has shown the ability to decrease colonization of undesirable bacteria such as Escherichia coli and Salmonella and increase the growth of desirable microorganisms. Prebiotics in poultry maintain normal intestinal microflora by competitive prohibition and antagonism, alter metabolism by decreasing bacterial enzyme activity and increasing digestive enzyme activity (Yaqoob et al., 2021; Jin et al., 2000; Islam et al., 2004). Plant extracts like flavonoids, carotenoids and other substances affect fatty acid accumulation in tissues and control the changes during meat storage (Koreleski and Swiatkiewicz, 2007; Ponnampalam et al., 2022).

The negative effects of antibiotics on health have encouraged scientists to find new and safe alternatives to these antibiotics, which may act as growth promoters with limited or no side effects. In this regard, an aqueous extract of C. intybus leaves may serve as a potential candidate. Therefore, this study was planned to examine the effect of aqueous extract of chicory leaves extracted at different pH on blood lipid and mineral profiles, feed conversion ratio and weight gain in broiler chicks.

Materials and Methods

The study was conducted at the Raja Muhammad Akram Nutrition Research Center, Institute of Animal Nutrition and Feed Technology, University of Agriculture, Faisalabad, to investigate the effect of C. intybus leaves extract on growth performance, blood lipids, and minerals profile in broilers. The various performance parameters, such as initial body weight, weekly body weight gain, weekly feed consumption, feed conversion ratio, daily water consumption, mortality, blood lipids (triglycerides, HDL, and LDL), and blood minerals (Ca, P, Na, and K) were recorded.

Preparation of chicory leaves extract

Fresh chicory/Kasni (Cichorium intybus) leaves were collected, sliced, meshed into small pieces, and dried under shade. The dried chicory leaves were ground into powder form. The leaf powder was mixed in distilled water @ 2 g / 100ml at three different pH levels, i.e., 3 pH (HCl), 7 pH (distilled water) and 12 pH (NaOH), respectively and then extracted at 80 °C for 3 h (Mavumengwana, 2004). After that, the extract was filtered and then neutralized with HCl or NaOH. These extracts were kept for 48 h at room temperature to be used for the experiment.

Experimental birds

One hundred fifty day-old-broiler chicks (Hubbard) of mixed sexes were purchased from a local hatchery. The chicks were randomly divided into 5 treatments with five replicates experimental units of 10 chicks each. These replicates were further allotted to five treatment groups positive control group (PC, diet with antibiotic Enracin, coccidiostat Salinomycin and fresh water without chicory extract), acidic chicory leave extract group (ACLE, diet without antibiotic but

 

Table I. Ingredient composition (%) and chemical composition of broiler experimental diets.

	Starter diet	Finisher diet
Ingredients
Corn	44.47	48.86
Millet	6.50	6.00
Rice polish	13.00	13.00
Wheat bran	7.50	5.20
Corn gluten 30%	2.50	2.50
Corn gluten 60%	2.00	2.50
Rapeseed meal	3.50	3.00
Soybean meal	9.83	8.40
Fish meal	7.50	7.50
Limestone	1.16	1.16
MDCP1	0.17	0.17
L-Lysine 78.8%	0.60	0.48
DL-Methionine 99.5%	0.25	0.21
L-threonine	0.05	0.05
Premix	0.30	0.30
Salt	0.067	0.067
Antibiotic (Enracin)	100 g/ton	100 g/ton
Coccidiostat (Salinomycin)	500 g/ton	500 g/ton
Total	100	100
Chemical composition
Protein %	19.65	18.75
ME (Kcal/Kg)	2850	2965
Fat %	5.26	5.31
Ash %	6.86	6.64
Lysine %	1.20	1.10
Methionine %	0.52	0.52
Ca%	0.98	0.90
Av. Phosphorus%	0.40	0.39
Crude fiber %	5.56	5.02

 

1Monodicalcium phosphate.

fresh water will contain chicory extract), neutral chicory leave extract group (NCLE, diet without antibiotic but fresh water will contain chicory extract), basic chicory leave extract group (BCLE, diet without antibiotic but fresh water will contain chicory extract) and negative control group (NC, diet without antibiotic and fresh water without chicory extract) in such a way that each treatment had three replicates in the trial. Nutrients composition is mentioned in Table I. On 35th day 3 ml blood was collected by wing vein puncture from three birds/replicate. Blood was allowed to clot in syringes (without anticoagulant), and serum was separated and stored in 1.5 ml Eppendorf tubes. Serum samples were stored refrigerated till submitted for analysis of minerals (Na, K, Ca and P) by atomic absorption spectrophotometry (Stef and Gergen, 2012) and lipids (triglycerides, LDL and HDL) by automatic biochemical analyzer Selectra (Jafari et al., 2011). Data collected was subjected to statistical analysis using the analysis of variance technique (ANOVA) under completely randomized design (CRD). Treatment means were compared by Duncan’s New Multiple Range tests (Steel et al., 1996).

Results and Discussion

Effect on performance characteristics

Table II shows that addition of various chicory leaves extract did not exhibit any significant (P<0.05) effect on total feed consumption of the birds of all treatment groups as compared to control groups (positive and negative). However, average feed consumption was numerically higher in the groups given water supplemented with chicory leaves extracts. These findings are in agreement with those observed by Asia and Gultekin (2012) who reported that feed consumption was not affected significantly in broilers treated with chicory leaves extract. It may be due to using chicory extracts in water instead of feed. Contrary to the results of the present study, Safamehr et al. (2013), Waldroup et al. (2013) and Behboud et al. (2011) found increased feed consumption in broilers treated with chicory by using different forms (pulp, powder) as feed additives and it may be due to its better palatability when mixed in feed.

The study results revealed that body weight gain was recorded significantly (P<0.05) higher in treatment BCLE, given water supplemented with chicory leaves extract, extracted at 12 pH, compared to other treatment groups. However, the difference in weight gain of the birds given water supplemented with chicory extracts, extracted at 3 pH, 7 pH, and antibiotic-supplemented feed was found to be non-significant (P>0.05). The higher body weight gain in birds treated with chicory leaves extract may be due to its complex carbohydrates such as oligofructose and oligosaccharides (Sangoh and Park, 2012; Chambers et al., 2011). The use of chicory has been shown to improve body weight gain, which probably is due to improved digestibility of nutrients by reducing microbial competition in the absorption of nutrients from host birds (Yousfi et al., 2017) and reducing ammonia production and other growth-depressing metabolites of microbes (Nabizadeh, 2012; Anderson et al., 2000).

 

Table II. Performance characteristics, blood lipids profile and blood minerals profile of broiler birds receiving different levels of aqueous extract of Cichorium intybus leaves.

Parameters	Treatment					SEM	P value
	PC1	ACLE2	NCLE3	BCLE4	NC5
Performance characteristics
Feed intake (g)	3060.23	3030.48	3000.76	3066.42	2982.89	31.01
Weight gain (g)	1818.73bc	1845.96ab	1791.23bc	1966.77a	1690.00c	29.27
FCR	1.68bc	1.64ab	1.67bc	1.56a	1.76c	0.02
Water intake	7100.66	7227.66	7368.00	7156.33	7026.00	54.97
Mortality %	0.66	0.33	0.66	0.66	0.33	0.13
Blood lipids profile
Triglycerides (mg/dl)	25.33b	30.00b	40.00a	38.33a	18.00c	2.26	0.00
HDL (mg/dl)	48.33	47.33	49.66	48.33	50.66	0.63	0.54
LDL (mg/dl)	64.66b	84.33a	68.00b	70.33b	40.66c	3.86	0.00
Blood minerals profile
Ca (mg/dl)	8.76	8.76	8.46	9.00	8.60	0.07	0.21
P (mg/dl)	5.40	5.27	5.23	5.46	5.64	0.09	0.75
Na (mmol/Lit.)	141.33	140.73	141.13	142.13	139.76	0.32	0.23
K (mmol/Lit.)	3.43	3.37	3.47	3.39	3.41	0.01	0.26

 

Values within the same row which have different superscripts are significantly different (P<0.05). 1Positive control, 2Acidic chicory leave extract, 3Neutral chicory leave extract, 4Basic chicory leave extract, 5Negative control.

 

These findings are similar to those observed by Silava et al. (2011), who reported increased body weight gain in broilers treated with chicory leaf extracts. The chicory leaves extract, at pH 12 showed better weight gain than those given water treated with chicory extracts, at pH3 and neutral pH. Maximum body weight gain due to chicory extract, at pH 12, may be due to the reduction in anti-nutritional factors of feed ingredients when treated with an alkaline solution of chicory, as observed by Wah et al. (1977). Chicory extracts may also have reduced pathogenic bacterial load in water and gastrointestinal tract, which improves the body weight gain by reduced intestinal thickness, which is very helpful in digestion and absorption of nutrients (Safamehr et al., 2013). Moreover, chicory has also been known to improve mucosal growth, villus height, width, crypt depth, and villus height to crypt depth ratio. These factors may stabilize nutrients and increase the digestion and absorption of these nutrients, thus enhancing body weight gain (Awad et al., 2011; Waldroup et al., 2013).

The addition of various C. intybus leaves extract in drinking water exhibited significant (P<0.05) effect on the FCR of the birds of the BCLE group as compared to PC, NCLE, and NC groups. The study’s results revealed significant (P<0.05) improvement in FCR of the birds in the treatment BCLE group, which were given water supplemented with C. intybus leaves extract, extracted at 12 pH as compared to those of other treatment groups except the ACLE group.

These findings are compatible with those observed by Silava et al. (2011), who reported improved FCR in broilers treated with C. intybus leaves extract. An improving trend was found in FCR due to the chicory leaves extract, at 12 compared to pH 3 and neutral pH levels. C. intybus extracts may have reduced pathogenic bacterial load in water and gastrointestinal tract which improved the body weight gain by reducing intestinal thickness and increasing intestinal length, which is very helpful in digestion and absorption of nutrients (Safamehr et al., 2013).

Mortality

The highest mortality was recorded in treatments PC, NCLE and BCLE groups (6.6%), followed by ACLE and NC groups (3.3%). Postmortem findings depicted that the cause of mortality was ascites in almost all the birds because of rapid growth, all these conditions require high oxygen levels in the blood (Kiiskinen, 1985). It was also observed that acidosis affects cellular membrane

 

Table III. Economic analysis of Broilers receiving different levels of aqueous extract of Cichorium intybus leaves.

Parameters	Treatment					SEM
	PC1	ACLE2	NCLE3	BCLE4	NC5
Total feed consumed (kg / bird)	3.06	3.03	2.99	3.06	2.98	0.03
Cost of total feed consumed/bird (PKR)	128.52	121.20	119.86	122.66	119.20	1.49
Cost/kg weight (PKR)	71.01c	65.85ab	67.03bc	62.62a	70.81c	0.96
Cost differential/kg gain (PKR)	0.00	5.15ab	3.98b	8.39a	0.19c	0.95
Relative cost benefit/kg gain (%)	100.00c	107.85ab	106.02bc	113.52a	100.38c	1.52

 

Values within the same row which have different superscripts are significantly different (P<0.05), 1Positive control, 2Acidic chicory leave extract, 3Neutral chicory leave extract, 4Basic chicory leave extract, 5Negative control.

 

integrity and reduce free radical elimination. Resultantly, leakage of blood vessels occurred and accumulated in the abdominal cavity, resulting in the development of ascites (Moschandreou, 2012).

Blood lipids profile

The NC group had the lowest (p<0.05) blood triglycerides, followed by the PC, ACLE, NCLE and BCLE groups. Higher serum triglyceride contents might be due to the increased digestibility of nutrients by the addition of chicory leaves extract, so as a result, more accumulation of triglycerides in the serum. Similar results were reported by Miao et al. (2008) in which they found increased levels of serum triglycerides at 0.6% and 1% inclusion levels of chicory leaves extract in broiler birds. In contrast, the results of Safamehr et al. (2013) were contrary to a recent study where they observed non-significant differences among all treatment groups with different chicory inclusion levels in bird’s diet. Asia and Gultekin (2012) found decreased serum triglycerides by the addition of chicory. Velasco et al. (2012) investigated a non-significance decrease in serum triglycerides by supplementing chicory root inclusions. Jafari et al. (2011) reported a significant decreasing trend of serum triglycerides when chicory was added to the feed. Navid and Mahmoud (2011) concluded that triglyceride concentrations were markedly reduced (p<0.05) in groups fed 1.5% and 2% chicory compared to the control, as shown in Table II.

The NC group had the lowest (p<0.05) blood triglycerides, followed by the PC, NCLE, BCLE, and ACLE groups. The results of Miao et al. (2008) favor our results, where they report an increased level of serum LDL at 0.6% and 1% inclusion of chicory extract in broiler birds. However, findings of Velasco et al. (2012) were in contrary to current study results in investigating a significant decrease in serum LDL with the supplementation of chicory root inclusions. Behboud et al. (2011) concluded that there was a decrease in the serum LDL with the supplementation of chicory and black seeds together. Jafari et al. (2011) observed a significant difference in serum LDL when chicory was added in the feed. Navid and Mahmoud (2011) concluded that serum LDL concentrations were markedly reduced (p<0.05) in groups fed 1.5% and 2% chicory compared to control.

Blood minerals profile

The BCLE group has higher (p>0.05) serum Ca concentration as compared to other treatment groups. The NC group has the highest (p>0.05) serum P concentration which is 5.64 compared to other treatment groups. Numerically maximum serum Na concentration was observed in BCLE group while the lowest in NC group. No significant difference was observed in serum K in all treatment groups. However, the maximum value was observed in the NCLE group, while the lowest value was observed in the ACLE group. The average values for Ca, P, Na, and K are given in Table II.

The results of the present study are in line with the findings of Safamehr et al. (2013), who reported that dietary inclusion of C. intybus was more beneficial in broiler production. Behboud et al. (2011) also reported a reduction in the cost of feed consumed at higher inclusion levels of C. intybus leaf pulp. However, in contrast, Liu et al. (2011) observed the high broiler production cost by including C. intybus as shown in Table III because of the high production, harvesting and processing cost of chicory leaves.

CONCLUSION

Based upon this study, it is concluded that replacement of AGPs with chicory leave aqueous extract, extracted at different pH revealed better performance of broilers in terms of weight gain, increased FCR, good assimilation of feed ingredients, low mortality rate, better feed intake along with better profit margin in comparison to control group of birds fed with untreated feed. Therefore, the use of chicory extract in broiler feed may be recommended as an inexpensive feed ingredient or additive, but an efficient alternative to AGPs as chicory leaves extract showed no pronounced side effects.

Funding

The authors extend their appreciation to Researchers Supporting Project number (RSPD2024R965), King Saud University, Riyadh, Saudi Arabia for funding this research.

IRB approval

The research was approved from Institutional Review Board.

Ethical statement

The rearing, sampling and culling of the birds were followed by the principles of Animal Ethical Committee of University of Agriculture, Faisalabad.

Statement of conflict of interest

The authors have declared no conflict of interest.

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