Effect of chives (Allium fistulosum L.) on growth performance and immune response of local chicken

Bui Thi Le Minh, Huynh Ngoc Trang, Luu Huu Manh, Nguyen Nhut Vinh Tu and Nguyen Nhut Xuan Dung

Veterinary Medicine Department, Agriculture and Applied Biology College, Cantho University, Vietnam
btlminh@ctu.edu.vn

Abstract

This study was conducted to evaluate the effect of dietary chive supplementation on growth performance, hematological and biochemical blood parameters, and fecal E.coli of growing chickens. One-day-old chickens (Noi Lai breed; n=120) were allocated to a completely randomized design with four replicates of four levels of fresh chives: 0, 1, 1.5 and 2% in the diet (DM basis).

There were linear improvements in growth rate and feed conversion of the chickens as the level of chives in the diet increased from 0 to 2% (DM basis) of the diet. There were no effects of chive supplementation on hematological and biochemical indices in blood. E. coli were shown to be susceptible to chive extracts (MIC test); however, supplementation with chives at up to 20g/kg diet DM had no apparent effect on fecal E. coli counts. It is suggested that chives could be considered as a prebiotic for natural growth promotion in chickens.

Key words: antimicrobial, E. coli, feed conversion, MIC, prebiotic

Introduction

Anti-microbial compounds are commonly used to control disease and to promote growth of poultry. However, the use of antibiotic growth promoters was banned by the European Union on 1^st January 2006, due to the risks of drug resistance in pathogenic bacteria. Many studies have been done to find alternatives to antibiotics in animal diets ( Guo et al 2000; Hertrampf 2001; Humphrey et al 2002) through the use of extracts of herbs and plants.

Chive is the common name of Alliumschoenoprasum L, belonging to the Allium species and Alliaceae family, and is in the list of plants proposed as feed additives in animal production. Allium species (garlic, onions, leeks and chives) contain organo-sulfur compounds such as the amino acids cysteine and methionine, γ-glutamyl peptides and especially S-substituted cysteine sulphoxides in varying amounts and proportions, corresponding to their flavor (FEEDAP 2005). Chives are herbaceous perennials with a strong onion or garlic scent, easy to grow and commonly used as afood additive and present in every Vietnamese kitchen. Chives have been well known for their medicinal properties, but there is very little information emphasizing their effect on growth performance of chickens.

The aims of this research were to evaluate the effect of increasing levels of dietary chives on feed intake, growth performance, blood parameters, and parasite infection in local chicken, The effects of a chive extract on E.coli were also studied.

Materials and methods

Location

The study was conducted on a farm in Binh Thuy ward, Cantho city and in the Veterinary Department of Cantho University, Vietnam, from June to September 2016.

Experimental design

Day-old chicks (Noi Lai breed; n = 320) were allocated in groups of 20 (equal male and female) to a completely randomized design with 4 treatments and 4 replicates. The experiment lasted 14 weeks. All birds were vaccinated against infectious diseases and had free access to feed and water. For the birds fed the control CH0 diet, their drinking water was diluted with Terra-colivet (Vemedim Corporation, Vietnam). This additive contained colistin, oxytetraxycline, vitamin A, D, E, K, B₂, B _5, B₆, B₁₂, PP, Na, K and dextrose).

The treatments were:

CH0: Basal diet with medicated water

CH1: Basal diet + 1% chives

CH1.5: Basal diet + 1.5% chives

CH2: Basal diet + 2% chives

The level of chives was based on their content of DM (9.2%). During the starter phase, the birds were fed the same basal diet. Introduction of the treatments (Table 1) began when the chicks reached 2 weeks of age. Fresh, whole chives (Photo 1) were purchased from a farm and chopped prior to adding them to the diets (Photo 2).

Hematologica lparameters

To evaluate the effects of dietary chives on blood, 64 birds were randomly selected (16 birds in each treatment) at the end of the experiment for hematology and plasma lipid analysis. Blood samples were taken from the wing vein in the morning before feeding using sterilized syringe and needles and EDTA anticoagulant-treated test tubes. The counts of red blood cells (RBC) and white blood cells (WBC) were determined by a hemocytometer method using the Natt-Herrick solution; hematocrit values were measured by microhematocrit methods (Kececi et al 1998).

To measure total cholesterol, HDL-cholesterol and HLD-cholesterol and triglycerides, plasma was collected by centrifugation at 3000 x g for 15 minutes. Measurements were done according to the colorimetric method using a biochemistry analyser (Siemens Advia 1200).

Isolation and identification of E. coli

Pooled cloacal swab samples (n=160) from 16 chickens were collected. The swab samples were and kept at 4^oC until microbiological examination. Bacteria were cultured in TBX medium (Tryptone-Bile X-glucuronide agar, Merck) at 37^oC for 24 h. The results were reported as number of E. coli per gram of feces.

Detection of parasite eggs in feces

Parasite eggs in feces (from hookworm, ascarid, whipworm, tapeworm and coccidia) were tested by the Modifed Wisconsin Sugar Centrifugal-Flotation Method, according to Pittman et al (2010).

Determination of E. coli minimal inhibitory concentration (MIC)

To confirm the antimicrobial activity, clean fresh chives were extracted by sterile distilled water in a 2:10 ratio; the mixtures were filtered through Whatman No. 4 filter paper and stored at 4^oC in a refrigerator until use. Ten E. coli colonies isolated from the chickens were used to determine the MIC of chive extract, according to the micro-dilution Broth method.

Photo 1. Preparing chives before feeding

Photo 2. Mixing chopped chives with the experimental feed

Feed analysis

The basal diet and chives were analyzed for dry matter, crude protein, crude fiber, ether extract, calcium and phosphorus according to the methods of AOAC (1995).

Data collection

Feed intake was recorded daily and live weight weekly. At the end of the study, measurements were made of: red and white blood cells and levels in blood of hematocrit, cholesterol, LDL-cholesterol, HDL-cholesterol and triglycerides.

Statistical analysis

The data were analysed using the General Linear Model procedure in Minitab (2001).

Photo 3. Chicks during starter phase	Photo 4. Feeding system

Feed intake, live weight gain and feed conversion

There was a curvilinear effect (R² = 0.99) on DM intake with the optimum observed for 1.5% of chives in the diet (Table 2; Figure 1). Growth rate was improved with a linear trend (R² = 0.96) and DM feed conversion by a curvilinear trend (R² = 0.92), as the level of chives in the diet was increased from 0 to 2% of diet DM (Table 2; Figures 1-3).

The initial increase in feed intake could be attributed to the smell/taste from the sulfur-containing components in chives that are thought to stimulate gustatory impulses (Bautista et al 2005). Gonzalez et al (2010) considered that the increase in feed intake observed with feeds containing onion came from the sweet taste and flavour attributed to the content of bispropenyl disulphide. Aditya et al (2017) also reported that onion ( Allium cepa) extract increased feed intake of chickens. However, the decline in feed intake with more than 1.5% of chives implies there is a limit to the sensory response with too high a concentration in the diet leading to reduced intake.

Figure 1. Effect of dietary chives on feed intake	Figure 2. Effect of dietqary chives on live weight gain	Figure 3. Effect of dietary chives on DM feed conversion

Part of the response in live weight gain induced by supplementation with chives could be the result of the increased feed intake. However, the feed intake response was curvilinear and decreased with more than 1.5% of chives in the diet (Figure 1). In contrast, the live weight gain response was linear (Figure 2) resulting in a positive response in DM feed conversion as the concentration of chives was increased from 1.0 to 2% of the diet DM (Figure 3).

Pharmacological effects of chives

Stajner et al (2004) and Rattanachaikunsopon et al (2008) postulated that the better growth performance of chickens fed chives may be related to the pharmacological effects from diallyl sulfides, flavonoids, vitamin C and carotenoids present in the chives. However, there was no evidence for this in the present study as there were no consistent effects of chive supplementation on hematological and biochemical parameters (Table 3). There were differences in tryglyceride levels in blood but these showed no consistent relationship with level of chives in the diet. Ademola et al (2009) found that supplementation of broiler chicken with garlic or ginger, alone or as a mixture, did not influence red blood cell counts or haemoglobin. Jimoh et al (2012) also reported that haematological parameters of broilers were not influenced by Allium sativum (garlic) dietary supplementation.

The chickens were healthy on all the diets with only one death (from coccidiosis) on each of the CH0, CH1 and CH1.5 diets.

Anti-microbial effects of chives

There were no differences among treatments in E.coli counts in feces, and no evidence of parasite eggs in the intestines (Table 4).

Minimal inhibitory concentration (MIC)

The test for antimicrobial activity showed that E. coli was susceptible to chive aqueous extracts (Table 5). The MIC of E. coli was in the range 25 -100 mg/ml (Table 4), indicating that chive extracts possess antibacterial potency.

Conclusions

• There were linear improvements in growth rate and feed conversion of local chickens as the level of chives in the diet increased from 0 to 2% (DM basis) of the diet.
  
  
• There were no effects of chive supplementation on haematological and biochemical indices in blood.
  
  
•  E. coli were shown to be susceptible to chive extracts (MIC test); however, supplementation with chives at up to 20g/kg diet DM had no apparent effect on fecal E. coli counts.
  
  
• It is suggested that chives could be considered as a prebiotic for natural growth promotion in chickens.

Acknowledgements

This study was supported by the Sida-financed project, MEKARN II. We thank the Department of Veterinary Medicine, Agriculture and Applied Biology College, Cantho University for providing facilities and a friendly environment to work. The authors are grateful to T R Preston for advice in writing this paper.

References

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Received 10 August 2017; Accepted 25 September 2017; Published 3 October 2017

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