Evaluation of Water Plantain (Alisma Canaliculatum A. Br. Et Bouche) And

Full Length Research Paper

Evaluation of water plantain (Alisma canaliculatum A. Br. et Bouche) and mistletoe (Viscum album L.) effects on broiler growth performance, meat composition and serum biochemical parameters

Md. Elias Hossain1, Gwi Man Kim1, Sang Soo Sun2, Jeffre D Firman3 and Chul Ju Yang1*

1Department of Animal Science and Technology, Sunchon National University, Suncheon 540-742, Korea. 2Department of Animal Science, Chonnam National University, Gwangju 500-757, Korea. 3Department of Animal Sciences, University of Missouri, Columbia, MO, 65211, USA.

Accepted 1 March, 2012

The present study was conducted to examine the potential use of water plantain (Alisma canaliculatum A. Br. et Bouche) and mistletoe (Viscum album L.) as alternative feed additives for broiler chickens. A total of 140 Ross broiler chicks were assigned to four dietary treatments over a five-week period. The dietary groups included; control (basal diet), antibiotic (basal diet + 0.005% oxytetracycline), water plantain (basal diet + 0.5% water plantain powder), and mistletoe (basal diet + 0.5% mistletoe powder). Results indicated that body weight gain and feed intake were not affected by the addition of water plantain and mistletoe to the diet. The feed conversion ratio (FCR) of the water plantain and mistletoe groups did not differ from the control group, although a better FCR was observed in the antibiotic group compare to the water plantain group. Crude protein as well as crude fat content of both breast and thigh meat in the water plantain group decreased, whereas crude protein content in breast meat was increased by the addition of mistletoe to the diet. When fatty acid composition was evaluated, a low n6/n3 ratio was found in the breast meat of the water plantain group, and supplemented groups exhibited a high polyunsaturated fatty acid content in thigh meat. The average thiobarbituric acid reactive substances values did not significantly change; however, breast meat of the water plantain group showed susceptibility to oxidation at Day 7. Serum biochemical data revealed that mistletoe had a hepatotoxic effect compare to water plantain, although both plants effectively enhanced immune function. It is suggested from the study results that water plantain could be incorporated into the diet of broiler chickens as an alternative feed additive.

Key words: Water plantain, mistletoe, feed additive, broiler.

INTRODUCTION

Plant extracts are ingredients of many commercial diet and reinforce immunity (Wenk, 2003). Since the ban on preparations currently used in animal production. They antibiotics in animal feed, producers often seek provide antioxidant (Cross et al., 2007), antimicrobial alternative natural feedstuffs to meet the demands of the (Manzanilla et al., 2004), immunity development (Ko et al., industry. 2008) and growth promoting effects (Lee et al., 2009). In Water plantain (Alisma canaliculatum A. Br. et Bouche) general, in the animal industry, herbs and other plant has many uses in traditional medicine; its roots are used extracts can help improve feed intake, digestive enzymes, to prevent diarrhea, hypertension, acute intestinal infection, mucous membrane dryness, dizziness, and jaundice in humans. It also reportedly suppresses DNA damage, has diuretic properties, and cholesterol lowering *Corresponding author. E-mail: [email protected]. Tel: 82-61-750- activities (Kim, 2003). Water plantain consists of 23% 3235. Fax: 82-61-750-3239. starch, 7% protein, and triterpene chemicals such as Hossain et al. 2161

alisol A, alisol B, alisol A monoacetate, alisol B replications with seven birds per replication over a period of five monoacetate, and epialisol A, which belong to the furfurol weeks, following a completely randomized design. The dietary and triterpenoid branches of refined oil. It also contains treatment groups included: Control (basal diet), antibiotic (basal diet + 0.005% oxytetracycline), water plantain (basal diet + 0.5% water D-glucose, D-fructose, sucrose, β-sitosterol and choline, plantain powder), and mistletoe (basal diet + 0.5% mistletoe with small amounts of alkaloids, aspartic acid, powder). All birds received a starter diet from 0 to 3 weeks and a phytosterols and their derivatives, palmitic acid, stearic finisher diet from 4 to 5 weeks. All diets were formulated to meet or acid, oleic acid, and linoleic acid (Jung, 1994; Science exceed the nutrient requirements of broiler chickens (NRC, 1994). Encyclopedia, 1991). The butanol fraction of Alisma has The ingredient composition and estimated nutrient content of the diets are shown in Table 1. been shown to reduce blood glucose levels, plasma triglycerides, free fatty acids, and cholesterol in rats (Kim, 2003). In addition, butanol with selenium supplementation Measurements and analysis increased liver glycogen and triglyceride levels, and reduced peroxidative liver damage in streptozotocin- Growth performance and meat composition induced diabetic rats (Choi, 2004). Antioxidant and Body weights were measured on a weekly basis from beginning to antidiabetic activities were also reported in rats (Choe et end of the experiment. Feed intake was determined by measuring al., 2008). feed residue on a weekly basis from the start of the experiment. Mistletoe (Viscum album L.) is abundant in forest Feed conversion ratio (FCR) was obtained by dividing the feed regions and orchards as a hemiparasite and has long intake by body weight gain. At the end of the experiment, broilers been used in animal diets especially during droughts and were slaughtered and samples were collected from breast and thigh winters. In comparison to commonly used conventional muscles. Moisture, crude protein, crude fat, and ash percentage of meat samples were analyzed according to Association of Analytical forages, mistletoe contains low protein, moderate fiber, Communities (AOAC, 2000) methods. and is high in minerals; therefore it can provide alternative mineral and forage sources for ruminant feeding (Madibela et al., 2000). The main constituents of Fatty acid composition and oxidative stability of meat mistletoe are lectins (mistletoe Lectins I, II, III), Fatty acid composition of breast and thigh meat was determined by viscotoxins, polysaccharides, cyclitols, flavonoids, phenyl the methyl ester extraction methods according to Yang et al. (2003). propane derivatives, triterpenoids like amyrin, betulinic Fatty acids were identified by matching their retention times with acid, oleanolic acid, phytosterols, amino acids, alkaloids, those of their relative standards (PUFA-2, Animal Source, Supelco, cyclic peptides, histamine, acetylcholine, and 9.3% Bellefonte, PA, USA) as well as by following the Food Composition protein (EMEA, 2000). Previous studies have Table (NRLSI, 2002). To determine the oxidative stability of broiler demonstrated that extracts from this plant possess breast and thigh meat, meat samples were preserved in the refrigerator at 4.5°C and thiobarbituric acid reactive substances pharmacological properties having immunomodulatory, (TBARS) values were assayed at Days 1, 3, 5 and 7 according to anti-inflammatory, cardiovascular, and antimicrobial Sarker et al. (2010b). TBARS values were expressed as micromole effects (Kienle and Kiene, 2003; Alison et al., 2000). of malondialdehyde (MDA) per hundred gram of meat. Nwaegerue et al. (2007) observed a glucose lowering effect in normal and diabetic rats using leaf extracts of V. album. It has also been shown to restore the suppressed Serum biochemical parameters immune response of fibrosarcoma-bearing mice, Blood samples were obtained at the end of the experiment by wing although cardiovascular effects in animal experiments are puncture and centrifuged at 3000 rpm for 20 min. Serum samples contradictory (Bissett, 1994; Jurin et al., 1997). Previous were analyzed for albumin, glutamyl oxaloacetic transaminase studies clearly concentrated mainly on cytotoxic, (GOT), glutamyl pyruvic transaminase (GPT), creatinine, blood urea cardiotoxic, immunostimulating, and antidiabetic nitrogen (BUN), total bilirubin, and total cholesterol with an automated clinical chemistry analyzer (FUJI DRI-CHEM 4000i, properties of water plantain and mistletoe in laboratory Japan) using commercially available kits. Serum IgG and IL-2 animals. Few attempts have been made to evaluate the concentration were determined in appropriately diluted samples by effect of these plants on broiler chickens (Sarker et al., a sandwich enzyme-linked immunosorbent assay (ELISA) using 2011; Kim et al., 2011; Kim et al., 2007) and pigs (Kim et microtiter plates and chicken specific ELISA quantitation kits for IgG al., 2010). Therefore, this experiment was designed to (Bethyl Laboratories Inc., USA) and interleukin 2 (Uscn Life Science evaluate the effect of water plantain and mistletoe on Inc. China). The ELISA procedure was carried out according to the protocol of the manufacturer and absorbance was measured with a growth performance, carcass composition, fatty acid Multiskan GO spectrophotometer (Thermo Fisher Scientific, composition, and oxidative stability of meat, as well as Finland). serum biochemical parameters and immunity in broilers.

Statistical analysis MATERIALS AND METHODS Data were analyzed using the general linear models of SAS Birds, feed, and experimental design Institute (2003) to estimate variance components with a completely randomized design. Duncan’s multiple comparison tests were used A total of 140 one-day old Ross broiler chicks were used for this to examine significant differences among the treatment means. The study. The chicks were assigned to four dietary treatments in five level of significance was set at p < 0.05. Data are presented as 2162 J. Med. Plants Res.

Table 1. Feed ingredients and chemical composition of broiler basal diets.

Items Starter (0-3 weeks) Finisher (4-5 weeks) Ingredients (g/kg, as feed basis) Corn grain 576.7 601.4 Soybean meal 265.0 254.0 Corn gluten 50.0 42.0 Soybean oil 22.0 15.0 Animal fats 45.0 50.0 Salt 2.50 2.50 Dicalcium phosphate 21.4 20.0 Limestone 9.2 8.8 Vitamin-Mineral premix1 3.0 3.0 Choline 0.8 0.7 L-lysine 2.4 1.6 Methionine 2.0 1.0

Calculated composition (g/kg, dry matter basis) Crude protein 220.0 200.0 Crude fat 40.0 45.0 Crude ash 80.0 80.0 Crude fiber 60.0 60.0 Methionine 7.9 7.0 Calcium 8.0 7.5 Available phosphorus 5.4 5.2 ME (MJ/kg) 12.98 13.19

1, Provided the following nutrients per kg of diet: vitamin A, 6000 IU; vitamin D3, 800 IU; vitamin E, 20 IU; vitamin K3, 2 mg; thiamin, 2 mg; riboflavin, 4 mg; vitamin B6, 2 mg; vitamin B12, 1 mg; pantothenic acid, 11 mg; niacin, 10 mg; biotin, 0.02 mg; Cu, 21 mg; Fe, 100 mg; Zn, 60 mg; Mn, 90 mg; I, 1.0 mg; Co, 0.3 mg; Se, 0.3 mg.

mean values ± standard error. crude protein content in thigh meat were also observed in the water plantain and mistletoe groups compare to the control group (p < 0.05). RESULTS AND DISCUSSION Water plantain and mistletoe are not commonly used in the livestock industry and very few documented studies Growth performances and meat composition were found to have evaluated their effects on broiler production. Plant extracts contain different molecules that Growth performances of broilers on different diet have intrinsic bioactivities affecting animal physiology and treatments are shown in Table 2. Body weight, weight metabolism. Some of these compounds have been gain, and feed intake were not different among the reported to improve animal performance due to their treatment groups (p > 0.05). FCR value of the water stimulating effect on salivation and pancreatic enzyme plantain group was higher (p < 0.05) compared to the secretions or by having a direct bactericidal effect on gut antibiotic group in the 4- to 5-week old chicks, but when microflora (Hardy, 2002). Body weights are commonly FCR was considered over the entire experimental period, used for monitoring the nutritional status and growth of the water plantain and mistletoe groups were not different animals (Ndlovu et al., 2007). Kim et al. (2011) reported a than the other groups. Addition of feed additives affected poor FCR in water plantain fed broilers during the the proximate composition of breast and thigh meat finishing period which is similar to our result, but their (Table 3). Moisture content of breast meat was findings for the total period were opposite of our results. decreased and crude protein content was increased in In addition, Sarker et al. (2010a) found that the use of the mistletoe group compared to the water plantain group water plantain probiotics had no negative effect on (p < 0.05). In addition, a lower crude fat content of breast growth and FCR in broilers. However, Kim et al. (2007) meat was found in the water plantain group compared to reported a better FCR when a 0.5% level of Korean the antibiotic group (p < 0.05). Higher moisture and lower mistletoe was fed to broilers. Previous investigators also Hossain et al. 2163

Table 2. Effect of diet treatments on broiler growth performance.

Parameter Treatment (week) Control Antibiotic Water plantain Mistletoe Body weight gain (g/bird) 0-3 626.32 ± 19.95 629.29 ± 24.74 632.83±27.86 619.89 ± 16.48 4-5 1252.75 ± 34.74 1255.36 ± 11.21 1262.54 ± 21.76 1236.96 ± 22.78 0-5 1879.07 ± 52.58 1884.64 ± 29.98 1895.36 ± 48.76 1856.86 ± 34.94

Feed intake (g/bird) 0-3 992.95 ± 27.42 967.86 ± 74.96 981.96 ± 21.19 972.68 ± 32.29 4-5 2156.18 ± 68.34 2055.43 ± 25.82 2150.32 ± 48.88 2077.82 ± 30.00 0-5 3149.13 ± 92.51 3023.29 ± 96.39 3132.29 ± 69.80 3050.50 ± 56.21

Feed conversion ratio (feed/gain) 0-3 1.59 ± 0.01 1.53 ± 0.07 1.56 ± 0.04 1.57 ± 0.03 4-5 1.72 ± 0.02a 1.64 ± 0.01b 1.71 ± 0.01a 1.68 ± 0.02ab 0-5 1.68 ± 0.01a 1.60 ± 0.03b 1.65 ± 0.02ab 1.64 ± 0.01ab a,b, Means with different superscripts in the same row differ significantly (p < 0.05).

Table 3. Effect of diet treatments on proximate composition of breast and thigh meat (g/100 g).

Treatment Parameter Control Antibiotic Water plantain Mistletoe Breast meat Moisture 73.84± 0.67ab 73.82 ± 0.56ab 74.83 ± 0.70a 72.33 ± 0.33b Crude ash 1.25 ± 0.04 1.25 ± 0.04 1.35 ± 0.11 1.51 ± 0.11 Crude fat 0.27 ± 0.02b 0.56 ± 0.11a 0.21 ± 0.03b 0.40 ± 0.04ab Crude protein 24.65± 0.67ab 24.37 ± 0.49ab 23.61± 0.66b 25.76 ± 0.19a

Thigh meat Moisture 74.50c ± 0.24 75.54 ± 0.43bc 77.15a ± 0.22 76.09 ± 0.48ab Crude ash 1.27 ± 0.05 1.18 ± 0.04 1.17 ± 0.03 1.29 ± 0.07 Crude fat 0.62 ± 0.16 0.66 ± 0.20 0.44 ± 0.07 0.56 ± 0.09 Crude protein 23.61 ± 0.13a 22.62 ± 0.49ab 21.24c ± 0.25 22.05 ± 0.49bc

a,b,c, Means with different superscripts in the same row differ significantly (p< 0.05).

found no differences in body weight gain or feed intake observed higher crude protein and lower moisture with the addition of water plantain and mistletoe to the content of breast meat in water plantain probiotic-fed diet which is consistent with our findings. Results of broilers. Inconsistent results were also observed in water phytobiotic application in broiler nutrition are not plantain and mistletoe probiotic-fed pigs (Kim et al., 2010) completely consistent. Some researchers stated and glasswort (Salicornia herbacea L.)-fed broilers significant positive effects on performance (Cross et al., (Sarker et al., 2010b). Plant extracts present some 2007; Perić et al., 2008), whereas another group reported advantages compared to traditional antimicrobials no influence on weight gain, food consumption, or because the latter have a single active principle. The conversion of feed (Ocak et al., 2008; Sarker et al., rapid metabolization and short half-life of active principles 2010b). Water plantain supplementation in our study in plants suggests that the risk of accumulation of these increased moisture and decreased crude protein content compounds in the tissue is negligible (Kohlert et al., of meat, in agreement with the results reported by Kim et 2000). Therefore, the presence of several compounds in al. (2011), while their findings in mistletoe group was one plant may have a synergistic beneficial effect in the opposite to our results. Contrarily, Sarker et al. (2010a) improvement of animal performance (Peres, 2007). 2164 J. Med. Plants Res.

Table 4. Effect of diet treatments on fatty acids composition of breast meat (g/100 g total fatty acid).

Treatment Fatty acids1 Control Antibiotic Water plantain Mistletoe Myristic acid (C14:0) 0.84 ± 0.03ab 0.96 ± 0.05 a 0.87 ± 0.01ab 0.81 ± 0.05b Palmitic acid (C16:0) 22.74 ± 0.04 23.40 ± 0.23 22.95 ± 0.50 22.69 ± 0.30 Palmitoleic acid (C16:1n7) 3.12 ± 0.40 2.74 ± 0.51 2.24 ± 0.04 2.89 ± 0.27 Stearic acid (C18:0) 5.49 ± 0.07 6.19 ± 0.29 6.36 ± 0.38 5.53 ± 0.36 Oleic acid (C18:1n9) 52.73 ± 0.81 53.33 ± 0.88 53.71 ± 1.15 53.30 ± 0.79 Linoleic acid (C18:2n6) 9.44 ± 0.21a 8.74 ± 0.33ab 8.33 ± 0.23b 9.10 ± 0.44ab α-linolenic acid (C18:3n3) 2.04 ± 0.15ab 1.48 ± 0.33 b 2.32 ± 0.24a 2.05 ± 0.19ab Eicosenoic acid (C20:1n9) 0.85 ± 0.03ab 0.74 ± 0.03b 0.75b ± 0.01 0.92± 0.09a Arachidonic acid (C:20:4n6) 0.72 ± 0.05 0.61 ± 0.07 0.69 ± 0.03 0.77 ± 0.05 Eicosapentaenoic acid (C20:5n3) 0.16 ± 0.01 0.15 ± 0.02 0.14 ± 0.01 0.14 ± 0.01 Adrenic acid (C22:4n6) 0.92 ± 0.13 0.74 ± 0.15 1.07 ± 0.10 1.05 ± 0.10 Docosahexaenoic acid (C22:6n3) 0.97 ± 0.02 0.94 ± 0.09 0.59 ± 0.19 0.76 ± 0.16 SFA 29.08 ± 0.06 30.55 ± 0.56 30.18 ± 0.86 29.03 ± 0.52 MUFA 56.69 ± 0.40 56.81 ± 0.36 56.69 ± 1.15 57.11 ± 0.53 PUFA 14.23 ± 0.49 12.64 ± 0.84 13.13 ± 0.43 13.86 ± 0.55 n3 3.16 ± 0.14 2.57 ± 0.34 3.05 ± 0.17 2.95 ± 0.14 n6 11.08 ± 0.39 10.08 ± 0.53 10.08 ± 0.28 10.91 ± 0.48 n6/n3 3.52 ± 0.13ab 4.04 ± 0.32a 3.32 ± 0.12 b 3.72 ± 0.20ab a,b, Values with different superscripts in the same row differ significantly (p < 0.05). 1, SFA = Saturated fatty acids; MUFA = monounsaturated fatty acids; PUFA = polyunsaturated fatty acids; n3 = omega 3 fatty acids; n6 = omega 6 fatty acids.

Table 5. Effect of diet treatments on fatty acid composition of thigh meat (g/100 g total fatty acid).

Treatment Fatty acids1 Control Antibiotic Water plantain Mistletoe Myristic acid (C14:0) 1.05a ± 0.01 1.06a ± 0.05 0.93b ± 0.01 1.04a ± 0.04 Palmitic acid (C16:0) 22.94 ± 0.21 23.18 ± 0.16 23.28 ± 0.26 23.64 ± 0.34 Palmitoleic acid (C16:1n7) 0.83b ± 0.05 1.16a ± 0.08 1.23a ± 0.10 1.33a ± 0.08 Stearic acid (C18:0) 8.02 ± 0.40 7.87 ± 0.24 7.31 ± 0.28 7.57 ± 0.25 Oleic acid (C18:1n9) 56.75a ± 0.52 55.13b ± 0.28 56.02ab ± 0.38 54.84b ± 0.52 Linoleic acid (C18:2n6) 7.12b ± 0.13 7.95a ± 0.19 7.54ab ± 0.21 7.59ab ± 0.29 α-linolenic acid (C18:3n3) 1.42 ± 0.11 1.63 ± 0.05 1.44 ± 0.16 1.55 ± 0.09 Eicosenoic acid (C20:1n9) 0.34b ± 0.01 0.43a ± 0.01 0.44a ± 0.24 0.51a ± 0.05 Arachidonic acid (C:20:4n6) 0.28b ± 0.02 0.30b ± 0.01 0.36ab ± 0.02 0.40a ± 0.05 Eicosapentaenoic acid (C20:5n3) 0.14b ± 0.01 0.19a ± 0.02 0.18ab ± 0.01 0.15ab ± 0.01 Adrenic acid (C22:4n6) 0.40 ± 0.06 0.44 ± 0.07 0.55 ± 0.10 0.63 ± 0.09 Docosahexaenoic acid (C22:6n3) 0.72 ± 0.02 0.68 ± 0.06 0.75 ± 0.04 0.76 ± 0.04 SFA 32.01 ± 0.60 32.11 ± 0.21 31.50 ± 0.25 32.24 ± 0.38 MUFA 57.91 ± 0.57 56.71 ± 0.36 57.69 ± 0.30 56.69 ± 0.42 PUFA 10.08b ± 0.28 11.18a ± 0.27 10.81ab ± 0.22 11.07a ± 0.37 n3 2.29 ± 0.13 2.50 ± 0.07 2.37 ± 0.15 2.46 ± 0.05 n6 7.80 ± 0.16 8.68 ± 0.21 8.44 ± 0.26 8.62 ± 0.40 n6/n3 3.43 ± 0.15 3.48 ± 0.05 3.61 ± 0.28 3.52 ± 0.22

a,b, Values with different superscripts in the same row differ significantly (p < 0.05). 1, SFA = Saturated fatty acids; MUFA = monounsaturated fatty acids; PUFA = polyunsaturated fatty acids; n3 = omega 3 fatty acids; n6 = omega 6 fatty acids.

Fatty acid composition of meat thigh meat are presented in Tables 4 and 5. Myristic acid content was lower (p < 0.05) in the breast meat of the Data concerning the fatty acid composition of breast and mistletoe group and thigh meat of the water plantain Hossain et al. 2165

Control Antibiotic Water plantain Mistletoe a 2.5 ab ab 2 b

1.5 1

0.5

MDA (µmol/100g MDA meat) (µmol/100g 0

Day 1 Day 3 Day 5 Day 7 Average

Figure 1. Effects of dietary water plantain and mistletoe on thiobarbituric acid reactive substance (TBARS) values of breast meat. TBARS values are expressed as micromole of malondialdehyde (MDA) per 100 g of meat. Data are presented as mean ± SE. Bars within a time class not sharing a common letter are significantly different (p < 0.05).

group compared to the antibiotic group. Linoleic acid not changed but a better n6/n3 ratio was found in the content of breast meat in the water plantain group was water plantain group for breast meat, although PUFA was decreased compared to the control, and α-linolenic acid increased in thigh meat. This finding is generally in line content was increased (p < 0.05) compared to the with the results of Koreleski and Swiatkiewicz (2007) and antibiotic group. Eicosenoic acid content of breast meat Ponte et al. (2008), who reported higher n3 fatty acid and was higher in the mistletoe group compared to the a lower n6/n3 ratio in breast meat of saga extract- and antibiotic and water plantain groups, while thigh meat of forage-fed broilers. Moreover, Sarker et al. (2011) found supplemented groups was higher (p < 0.05) compared to improvement of PUFA in breast and thigh meat and the control. Palmitoleic acid content of thigh meat was n6/n3 ratio in thigh meat of a water plantain probiotic increased in the supplemented groups compared to the group. However, no differences were reported in the SFA, control, whereas eicosapentaenoic acid content was MUFA, and PUFA content of both water plantain- and significantly increased (p < 0.05) in the antibiotic group mistletoe-fed broiler and pig meat (Kim et al., 2011; Kim compared to the control group. Oleic acid content of thigh et al., 2010). meat was decreased and arachidonic acid was increased (p < 0.05) in the mistletoe group compared to the control and antibiotic group, respectively. The water plantain Oxidative stability of meat group presented a lower n6/n3 ratio in the breast meat, whereas higher polyunsaturated fatty acid (PUFA) TBARS values of meat were evaluated and no content was found in the thigh meat of the supplemented differences were observed among the treatments except groups (p < 0.05). From the aspect of human health, the at Day 7 in the refrigerated breast meat and Day 5 in the fatty acid composition of meat products is an important thigh meat (Figures 1 and 2). In breast meat, the water parameter of meat quality. With the recognition that the plantain group showed a higher value (p < 0.05) groups of PUFA each play their own metabolic roles in compared to the antibiotic group at Day 7, but did not health promotion, the role of the PUFA/SFA ratio has differ from the mistletoe group. The antibiotic group also decreased while the importance of the total n6/n3 ratio had a lower (p < 0.05) value than the control group at has increased (Dublecz et al., 2004). PUFA content of Day 5 in thigh meat, but no differences were observed modern diets are low in n3 fatty acids leading to high among the additive groups. Results from TBARS testing n6/n3 fatty acid ratios, which are estimated at 25:1 to revealed that breast meat from broilers fed water plantain 50:1 in western countries instead of the recommended 1- may be more susceptible to oxidation than that from 4:1 (Grashorn, 2007). For these reasons, many antibiotic-fed broilers. These results are in agreement production facilities have sought ways to change the fatty with data reported by Koreleski and Swiatkiewicz (2007), acid composition of meat, mainly through feeding plant who stated that plant extracts did not favorably affect sources of PUFA. In our experiment, PUFA content was TBARS value in broiler meat. A higher TBARS content 2166 J. Med. Plants Res.

Control Antibiotic Water plantain Mistletoe 7 6 5 a 4 ab ab 3 b 2 1

MDA (µmol/100g MDA meat) (µmol/100g 0

Day 1 Day 3 Day 5 Day 7 Average

Figure 2. Effects of dietary water plantain and mistletoe on thiobarbituric acid reactive substance (TBARS) values of thigh meat. TBARS values are expressed as micromole of malondialdehyde (MDA) per 100 g of meat. Data are presented as mean ± SE. Bars within a time class not sharing a common letter are significantly different (p < 0.05).

Table 6. Effect of diet treatments on serum biochemical parameters.

Treatment Parameters1 Control Antibiotic Water plantain Mistletoe Albumin (g/dl) 3.13 ± 0.09b 3.37 ±0.12ab 3.49 ± 0.06ab 3.68 ± 0.19a GOT (u/L) 48.00 ± 6.93b 45.33 ± 3.71b 55.00 ± 4.04ab 67.33 ± 6.5 7a GPT (u/L) 14.67 ± 3.28b 15.33 ± 1.76b 16.00 ± 1.16b 24.67 ± 3.48a Creatinine (mg/dl) 0.83 ± 0.09 0.83 ± 0.13 0.73 ± 0.04 0.82 ± 0.02 BUN (mg/dl) 8.17 ± 0.65c 7.00 ± 0.49c 12.27 ± 0.24b 14.20 ± 0.26a Bilirubin (mg/dl) 0.43 ± 0.09ab 0.37 ± 0.09ab 0.34 ± 0.04b 0.60 ± 0.06a Cholesterol (mg/dl) 154.33 ± 9.35 142.67±12.71 146.67± 14.42 153.67± 24.84 IgG (mg/dl) 4.01 ± 0.11 4.11 ± 0.06 4.07 ± 0.08 4.14 ± 0.04 IL2 (pg/ml) 93.67 ± 35.30b 73.75 ± 34.00b 305.34 ± 22.16a 263.22 ± 28.67a

a,b,c, Values with different superscripts in the same row differ significantly (p < 0.05). 1, GOT = Glutamyl oxaloacetic transaminase; GPT = glutamyl pyruvic transaminase; BUN = blood urea nitrogen; IgG = Immunoglobulin G; IL2 = Interleukin 2.

was noted in refrigerated breast and thigh meat at Day 5 Ko et al., 2008) have demonstrated the principal potential from broilers whose diet was supplemented with water of feed additives from the plant family containing plantain probiotics, although no difference was observed bioactive compounds to improve the oxidative stability of at the end of Day 7 (Sarker et al., 2011). Tanabe et al. animal derived products. Therefore, it is suggested that (2002) compared the antioxidant activities of 22 culinary plant extracts, directly added to feed, may influence the herbs and spices commonly used for pork meat and oxidative stability of meat after slaughter. found variations of antioxidant activity among them. The antioxidant activity of plant extracts is mainly related to the presence of phenolic compounds. Moreover, Serum biochemical parameters compounds such as flavonoids and terpenoids also present antioxidant activity (Rizzo et al., 2008). These The effect of including water plantain and mistletoe as substances may intercept and neutralize free radicals, feed additives on serum biochemical parameters of preventing the propagation of the oxidation process. broilers are illustrated in Table 6. Data revealed that the Several recent studies in poultry (Florou-Paneri et al., serum albumin, GOT, GPT, BUN, and bilirubin content 2006; Schiavone et al., 2007) and pork (Janz et al., 2007; were higher in the mistletoe group (p < 0.05). The water Hossain et al. 2167

plantain group had a lower GPT, BUN, and bilirubin plantain and mistletoe suggests that they have content compared to the mistletoe group (p < 0.05). immunomodulatory effects. This result was in harmony There were no significant differences in creatinine and with the findings of Kim et al. (2011) and Tuan et al. cholesterol content among the groups. Dietary (2009), who reported that water plantain, mistletoe, and supplementation of water plantain and mistletoe markedly purple coneflower [Echinacea purpurea (L.) Moench] increased (p < 0.05) serum IL-2 production, although IgG extract significantly enhanced IL-2 production in broilers. concentration was not significant. The level of additives applied in our study may be Serum biochemistry can reflect the condition of an responsible for the differences in effect seen. Although organism and the changes happening to it under the the active principles are responsible for the main effect influence of internal and external factors. Liver enzymes on the animal, research studies show the presence of a GOT and GPT can be measured in serum as markers of synergistic effect among plant primary and secondary hepatic damage. The observations of no significant components with the secondary components enhancing changes in liver enzymes and bilirubin levels, in the effects of the primary components (Kamel, 2001). comparison to control, may indicate that water plantain More studies are necessary to evaluate the had no negative effect on liver functions. The results hematological, immunological, and antimicrobial activities coincide with Kim et al. (2011), who found the same in of these plants. water plantain-fed broilers. Previous researchers also reported hepatoprotective effects in response to feeding different medicinal plants to broilers (Al-Jaff, 2011; Chand Conclusions et al., 2011). Serum concentration of these enzymes and bilirubin were increased by the toxic effect of mistletoe on Growth performance was not altered by the addition of the liver resulting in necrosis or changes in cell water plantain and mistletoe to the diet of broilers, membrane permeability, which can be associated with although crude fat and crude protein content of the meat liver dysfunction. Abdelati et al. (2008) also reported a were lower in the water plantain group. The dietary higher level of GOT in broilers fed white leadtree supplementation of water plantain and mistletoe [Leucaena leucocephala (Lam.) de Wit] seeds. Similarly, positively affected the fatty acid composition of the broiler increased AST was observed in broilers fed essential oil, meat. No antioxidative effect was observed in the pepper (Capsicum frutescens L.) extract, and rosemary supplemented groups, but the breast meat of the water (Rosmarinus officinalis L.) leaf meal (Ghazalah and Ali, plantain group showed more susceptibility to oxidation. 2008; Traesel et al., 2010). This therefore, suggests that Among the medicinal plants used in this study, mistletoe the hepatotoxic effects of mistletoe may be due to its had a hepatotoxic effect as compare to water plantain, bioactive components. while both plants effectively enhanced immune function. Albumin is the protein with the highest concentration in the plasma. Increased serum albumin in the mistletoe group was consistent with the findings of Al-Jaff (2011), ACKNOWLEDGMENTS whereas the finding of no effect in the water plantain group was also similar to previous results (Abdelati et al., The authors would like to thank the Korea Institute of 2008; Traesel et al., 2010). 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