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Effects of Flavonoids from Allium Mongolicum Regel on Growth Performance and Growth-Related Hormones in Meat Sheep

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Effects of Flavonoids from Allium Mongolicum Regel on Growth Performance and Growth-Related Hormones in Meat Sheep Accepted Manuscript Effects of flavonoids from Allium mongolicum Regel on growth performance and growth-related hormones in meat sheep Muqier, Sarula Qi, Terigele Wang, Renwei Chen, Cuifang Wang, Changjin Ao PII: S2405-6545(16)30156-1 DOI: 10.1016/j.aninu.2017.01.003 Reference: ANINU 133 To appear in: Animal Nutrition Journal Received Date: 29 August 2016 Accepted Date: 10 January 2017 Please cite this article as: Muqier Qi S, Wang T, Chen R, Wang C, Ao C, Effects of flavonoids from Allium mongolicum Regel on growth performance and growth-related hormones in meat sheep, Animal Nutrition Journal (2017), doi: 10.1016/j.aninu.2017.01.003. This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. ACCEPTED MANUSCRIPT 1 Effects of flavonoids from Allium mongolicum Regel on growth performance and 2 growth-related hormones in meat sheep 3 4 Muqier, SarulaQi, Terigele Wang, Renwei Chen, Cuifang Wang, ChangjinAo* 5 6 College of Animal Science, Inner Mongolia Agricultural University, Hohhot 010018, 7 China 8 9 10 * Corresponding author. 11 E-mail address: [email protected] (C. Ao) 12 13 14 15 16 MANUSCRIPT 17 18 19 20 21 22 23 24 25 ACCEPTED 26 27 28 29 30 ACCEPTED MANUSCRIPT 31 ABSTRACT 32 This study was conducted to investigate the effects of different doses of flavonoids 33 from Allium mongolicum Regel on the production performance and neuroendocrine 34 hormones in meat sheep and to determine the optimum dosage of Allium mongolicum 35 Regel flavonoids to add to the basal diet of dry lot-feeding meat sheep. Sixty meat 36 sheep (initial body weight = 39.9 ± 3.2 kg; 6-month-old) were randomly assigned to 4 37 groups (15 sheep per group). The sheep in the control group were fed a basal diet, and 38 the 3 experimental groups were fed the basal diet supplemented with flavonoids at 11, 39 22 and 33 mg/kg. Blood samples were collected via the jugular vein at d 0, 15, 30, 45, 40 and 60 to determine the neuroendocrine hormone levels. The fasting weight of the 41 sheep was measured during the experimental period, and feed offered and refusals 42 were recorded daily. The basal diet supplemented with flavonoids from 11 to 33 mg/kg 43 significantly increased the daily weight gain and average daily feed intake (P < 0.05) 44 and significantly decreased the feed conversion ratio (P < 0.05), but there were no 45 differences among the supplementation groups (P > 0.05). Starting on d 30, the growth 46 hormone (GH) and insulin-like growth factor-IMANUSCRIPT (IGF-I) levels in the sera of the sheep in 47 the supplementation groups increased significantly ( P < 0.05), and the increases 48 occurred in a time-dependent manner. Compared with control group, after d 30, the 49 serum corticosterone (CORT) levels were reduced in the sheep that consumed the basal 50 diet supplemented with 22 mg/kg flavonoids ( P < 0.05), but among the other 51 experimental groups, there was a non-significant effect ( P > 0.05). The serum 52 adrenocorticotropic hormone (ACTH) levels were increased by the supplementation of 53 flavonoids, but compared with the control group, the effect was not significant. The 54 basal diet supplemented with flavonoids at levels of 11 to 33 mg/kg had a significant 55 effect on theACCEPTED production performance and neuroendocrine hormone levels of meat 56 sheep, and the effect occurred in a time-dependent manner. The effect was especially 57 obvious after 30 d of feeding. 58 Key words: Allium mongolicum Regel , Flavonoids, Growth performance, Hormones, 59 Meat sheep ACCEPTED MANUSCRIPT 60 1 Introduction 61 Allium mongolicum Regel , also known as the Mongolia leek, is a type of 62 Liliaceous allium plant that grows in high altitude desert steppe and desert areas; it is 63 perennial and xerophytic (Wang et al., 2013). It is mainly found in desert land in 64 Qinghai, Gansu, Xinjiang and Inner Mongolia, and is especially abundant in the 65 western areas of Xilingol, Ordos, and Alxain Inner Mongolia. Allium mongolicum 66 Regel, as a characteristic wild vegetable, is a natural and healthy food with a high 67 nutritional value, unique flavour and supreme palatability (Chen et al., 2000). Allium 68 mongolicum Regel and its extracts have been shown to increase the average daily gain 69 (ADG), feed intake and feed remuneration of broiler chickens (Ha, 2008) and to 70 significantly affect growth-related hormones (Zhang, 2005). 71 Flavonoids are active ingredients in natural plants that can promote growth, 72 neuroendocrine and immune function of animals. Related research found that the 73 isoflavone daidzein promoted animal growth (Han, 1999), improved immune function 74 (Guo et al., 2004), and enhanced lactation and laying (Hu and Zhang, 2009; Yang, 75 2006). Flavonoids are an important active substanceMANUSCRIPT in Allium mongolicum Regel (Sa, 76 2014a). According to research, Allium mongolicum Regel flavonoids show 77 biological efficacy as anti-oxidants and have anti-bacterial and immune regulation 78 abilities. Allium mongolicum Regel provides small doses, low toxicity, and high 79 efficacy in practical application; consequently, it offers good latent capacity for new 80 types of feed additives. Animal growth is a complicated metabolic process influenced 81 by genes, nutrients, hormones and the environment and regulated by neuroendocrine 82 function (Tao, 2006), of which the hypothalamic-pituitary-growth axis is the key 83 regulator(Sarah, 2003). 84 Currently,ACCEPTED research on the flavonoids from Allium mongolicum Regel has mainly 85 focused on its in vitro anti-oxidant, anti-bacterial and anti-viral activities; studies of 86 growth performance and growth-related hormones in meat sheep are rare. On the basis 87 of a preliminary in vitro study (Bao, 2015; Sa, 2014b), this study selected the 88 flavonoids from Allium mongolicum Regel as feed additives for meat sheep. Different 89 doses of flavonoids from Allium mongolicum Regel were added to the basal diet, and ACCEPTED MANUSCRIPT 90 the effect on growth performance and growth-related hormones in meat sheepwas 91 explored. A suitable amount of flavonoids from Allium mongolicum Regel was 92 determined for use as a natural feed additive. 93 94 2 Materials and methods 95 2.1 Experimental flavonoids from Allium mongolicum Regel 96 The flavonoids from Allium mongolicum Regel were extracted using the 97 extraction processing method described by Saruli (Sa, 2014b). An ultrasonic extraction 98 method was adopted using the following parameters: extraction time = 15 min, 99 extraction temperature = 40 °C, alcohol density =75%, and material-to-liquid ratio = 100 1:30. The projected total flavonoid yield was 12.85 mg/g. The flavonoids from Allium 101 mongolicum Regel are yellow in colour and insoluble in water. According to early 102 research regarding structure identification (Sa, 2014b), the structure contains 103 saccharides, naphthenic hydrocarbon,3',4' -epoxygroup-7-0-5-methoxy flavonols, 104 7-0-5,4'-dimethoxy-3'-hydroxide radical flavones, rutin, quercitrin, saccharides, 105 mignonette element - 5 0 - glucose - 4 - hydroxybenMANUSCRIPTzoic acid, and acacia. 106 2.2 Reagents and apparatus 107 Testing kits were used to determine the levels of serum growth hormone (GH; 108 HY-C0018), serum insulin-like growth factor-1 (IGF-1; HY-H0024), serum 109 corticosterone (CORT; HY-D0032), and serum adrenocorticotropic hormone (ACTH; 110 HY-D0023). The testing method was radioimmunoassay, and the test apparatus was an 111 r-911 auto radio free counter (provided by the Science and Technology Industrial 112 Company of the University of Science and Technology of China). All 113 reagents and apparatuses were provided by the Beijing Sino-UK Institute of Biological 114 Technology.ACCEPTED 115 2.3 Experimental method 116 2.3.1 Selection of experimental animals 117 A total of 60 six-month-old healthy meat sheep with initial body weight of 39.9 ± 118 3.2 kg were divided into 4 groups of 15 sheep, which included the control group, test 119 group1, test group 2 and test group 3. ACCEPTED MANUSCRIPT 120 2.3.2 Experimental diet 121 The control group was fed a basal diet, and the 3 test groups were supplemented 122 with 11, 22, and 33 mg/kg flavonoids from Allium mongolicum Regel . According to the 123 stability test of flavonoids from Allium mongolicum Regel in the rumen, adding 22 124 mg/kg of total flavonoids from Allium mongolicum Regel to the rumen could improve 125 parameters related to rumen microbial fermentation gas production, pH, ammonia 126 nitrogen, microbial protein, and volatile fatty acids and could optimize the effects of 127 the flavonoids (Bao, 2015). This study referred to these previous results when choosing 128 the supplementation doses (11, 22, and 33 mg/kg) of flavonoids from Allium 129 mongolicum Regel . The components of the experimental diet testing method (Zhang, 130 2003) were as follows: 1) The crude protein of the feedstuff was tested using the 131 Kjeldahl Nitrogen Determination method; 2) The Van Soest detergent fibre analysis 132 method was used for the determination of neutral detergent fibre (NDF) and acid 133 detergent fibre (ADF) contents; 3) The calcium content of feedstuff was tested using 134 potassium permanganate titration; 4) The phosphorus content was determined using 135 colorimetry with molybdenum yellow. The basicMANUSCRIPT dietary composition and nutritional 136 levels are shown in Table 1. 137 2.3.3 Feeding management 138 The entire experimental period lasted 75 d, and the adaptation period lasted 15d.
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