Effect of Spirulina-Chinese herbal medicine on quality of eggs and serum biochemical indexes of egg-laying hens

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1 B- 640 [1-5] Indian J. Anim. Res., rint ISSN: / Online ISSN: AGRICULTURAL RESEARCH COMMUNICATION CENTRE Effect of Spirulina-Chinese herbal medicine on quality of eggs and serum biochemical indexes of egg-laying hens Liu eipei and Zang Sumin* College of Animal Science and Technology, Hebei Agricultural University, Baoding , Hebei China Received: Accepted: DOI: /ijar.v0iOF.9143 ABSTRACT This study used three-factor linear regression orthogonal design to examine the impacts of traditional Chinese herbal medicine on egg quality and egg-laying hens biochemical indexes. 495 and 210d Hy-Line gray commercial egg-laying hens were selected and spirulina powder was added to their diet at 5 g/kg, 10g/kg, 15g/kg in the diet, rchmannia glutnosa polysaccharides were added at 2.5g/kg, 3 g/kg, 3.5 g/kg, and Leonurus Japonicas was added at 0.7 g/kg, 0.9g/kg, 1.1 g/kg. The hens were randomly allocated to a control group and ten test groups, in order to study the herbs effects on laying performance and immunity parameters. The results showed that: (1)When spirulina-chinese herbal medicine was added, the average egg weight, yolk color, and Hartz unit increased significantly.(2) When spirulina powder levels were increased, the egg yolk color and iron content significantly increased while the cholesterol and triglyceride levels were significantly lower (<0.05). With the increase of rchmannia glutnosa polysaccharides extracts, eggshell strength increased significantly (<0.01). Egg yolk color and eggshell strength increased significantly (<0.05 and <0.01, respectively) when Leonurus Japonicas increased. (3) Spirulina powder and rchmannia glutnosa polysaccharides extracts were significantly antagonistic to average egg weight and eggshell strength (<0.01). Leonurus Japonicus and rchmannia glutnosa polysaccharides were significantly antagonistic to eggshell strength and cholesterol levels (<0.05). Key words: Biochemical indices, Egg quality, Hens, Leonurus Japonicas, RS, Spirulina powder. INTRODUCTION Spirulina powder is an herb used in traditional Chinese medicine to enhance the quality of eggs laid by hens. It is low in fat and cholesterol, and has high levels of chlorophyll, carotene, vitamins, and trace elements (Qi, 2010 and Zheng, 2009). As it contains polysaccharide, phycocyanin, and - carotene, spirulina powder is able to promote serum protein biosynthesis. Ground yellow polysaccharide extract is also used in traditional medicine, and contains rchmannia glutnosa polysaccharides, which have good pharmacological effects because they allow for adequate levels of sugars, amino acids, and iridoid glycosides (Li et al., 2006). Another traditional herb is Leonurus Japonicas (motherwort), which can affect calcium and phosphorus metabolism, which may affect the quality of eggs laid by hens because of its chemical composition of alkaloids, diterpenoids, and flavonoids (Luo et al., 1986; Zheng et al., 1997; Miao et al., 2001 and Wang et al., 1991). Leonurus japonicus is able to reduce cholesterol and triglycerides, but to date; most research has focused on its effects on uterine contractions (Yang et al., 2001; Ma et al., 2000 and Chan et al., 1983). Nowadays, there had not been research on the effects of rehmannia polysaccharide on egglaying hens. This experiment analyzed the effects of different amounts of spirulina powder, ground yellow polysaccharide extract, and Leonurus Japonicus on egg quality and serum biochemical parameters and we then determine the appropriate amount and the interaction effect between them by adding each in various amounts. We then select the best formula. MATERIALS AND METHODS For this experiment, 495 and 210d Hy-Line gray commercial layer hens were selected and randomly assigned to 11 groups, using three-factor linear regression orthogonal design, as shown in Table 1. This was repeated three times with 15 hens per replicate. Basic level of the diet with reference to the industry standard of the eople s Republic of China standard recommend laying hens nutrition standards(2004)and NRC(1994).The compositions of the experimental diets are presented in Table 2. The hens were fed three times per day. Eggs were collected daily and weighed. At the last feeding trial, 8 eggs were collected per dietary treatment and egg quality was measured using the average egg weight (g), egg yolk color, hartz unit, eggshell strength, eggshell thickness, and egg-shaped index. Before the end of the feeding trial day, blood samples *Corresponding author s suminzang@163.com

2 2 INDIAN JOURNAL OF ANIMAL RESEARCH Table 1: Test design No. Z0 Z1 Z2 Z1Z2 Z3 Z1Z3 Z2Z3 Z1Z2Z (5) -1(2.5) 1-1(0.7) (1.1) (3.5) (15) (10) 0(3) 0 0(0.9) control group Note: Table Z1, Z2, Z3 are respectively spirulina powder, rchmannia glutnosa polysaccharides and Leonurus Japonicas of standardized variable material; units are g/kg,; - indicates 0; (1,0,-1 is the encoded factor in the orthogonal test, with 1 representing the upper level, 0 the zero level, -1 the Lower level). Table 2: Composition of experimental diets Ingredient (%) Nutrient levels Corn Metabolic energy/(mj/kg) Soybean meal Crude protein/% Stone powder 8.20 calcium/% 3.54 Dicalcium phosphate 1.57 effective phosphorus/% 0.38 remix * 1 lysine/% 0.83 Salt 0.31 methionine/% 0.27 Total 100 *er kilogram of diet: Iron 60.00mg, copper 8.00mg, magnesium 70.00mg, Mn 60.00mg, zinc 80.00mg, iodine and selenium 0.35mg 0.30 mg, vitamin A IU, vitamin D IU, vitamin E 5.00IU, vitamin K 0.50IU, vitamin B1 0.80IU, vitamin B2 2.50mg, vitamin and vitamin B6 3.00mg,B mg, pantothenic acid 2.20mg, niacin 20.00mg, biotin and folic acid 0.10mg 0.25mg. were collected from the wing vein of ten chicks from each group to determine calcium, iron, total protein, cholesterol, and triglyceride levels. RESULTS AND DISCUSSION The changes in average egg weight (g), egg yolk color, Hartz unit, eggshell strength, eggshell thickness, and egg-shaped index are shown in Table 3. When the traditional Chinese herbal medicine additive spirulina was present, the average egg weight and Hartz unit linearly decreased and the yolk color increased significantly in comparison to the control group (<0.05). Group 6 had the highest increase of all of the groups. In this group, the average egg weight increased by 3.62% over the control group. The Hartz unit of group 9 increased by 9.04% over the control group (<0.05). Group 4 of eggshell strength and eggshell thickness was the highest that the eggshell strength increased by 15.32% (<0.05) and eggshell thickness increased by 16.67% (<0.05) over the control group. When levels of spirulina powder or rchmannia glutnosa polysaccharides extract were increased, the average egg weight decreased significantly (<0.01), egg yolk color significantly increased (<0.05), and egg shape index increased (<0.1). With the increase of rchmannia glutnosa polysaccharides, the eggshell strength increased significantly (<0.01) and egg yolk color increased (<0.1). With the increase of Leonurus japonicus, there was a significant increase in the average egg weight (<0.05), and a significant increase in eggshell strength (<0.01). When average egg weight and eggshell strength are used as an indicator, spirulina powder and rchmannia glutnosa polysaccharides extracts showed significant antagonism ( <0.01). Leonurus japonicus and spirulina powder antagonism had significant changes in eggshell strength, ( <0.05). Table 5 shows that when the traditional Chinese herbal medicine additive spirulina was added to the diet of egg-laying hens, their cholesterol and triglyceride levels decreased significantly effect in comparison to the control group (<0.05). Group 8 of the cholesterol and triglyceride levels had the most significant difference when compared to the control group (<0.01), which had the lowest. There was no significant impact on levels of iron and albumin. The calcium content of Group 9 increased by 12.69% when compared with the control group (<0.01). We can see form Table 6 showed that when the level of spirulina powder was increased, the cholesterol and triglyceride content was significantly reduced and the Iron content increased significantly (<0.05). With the increase of Leonurus japonicus, triglycerides tended to decrease ( <

3 0.1). Leonurus japonicus and rchmannia glutnosa polysaccharides had an antagonistic effect on cholesterol levels (<0.05). With the addition of spirulina, the average egg weight, yolk color and Hartz unit were enhanced, and cholesterol and triglyceride levels were significantly lower (<0.05). These results are in agreement with the findings of previous research from Dai, Ning, Ge and Xu. Their research (Dai et al.,, 2012) showed that egg yolk color was significantly improved after spirulina powder was fed to egg-laying hens instead of the same amount of fish meal. Layer itself couldn t synthetic pigment and only obtained from the feed (Cheng et al., 2001). The increase of eggshell color was related to chlorophyll and carotene in Spirulina powder. Ning et al. (2004) found that when 2% spirulina powder was added to the basal diet of egg-laying hens, egg weight significantly increased (<0.05), and there was significant improvement in yolk color. A similar phenomenon was reported by Ge et al. (1990) and Xu et al. (1990). The results showed that spirulina powder could improve egg quality. Large doses of spirulina could prevent an increase in liver triglyceride levels caused by fructose and reduce blood cholesterol levels. It Vol. Issue, () Table 3: Effect of different levels of spirulina - Chinese herbal medicine additive on egg quality of egg-laying hens No. Egg weight Egg yolk color Hartz unit Eggshell strength EggshellThickness(mm) Egg-shaped index ±0.03ab 9.83±0.408b 80.12±3.28BCbc 35.79±4.61ABabc 0.374±0.026a 1.285±0.048ab ±0.01ab 9.83.±0.408b 77.75±6.39ABCb 33.69±2.55Aa 0.378±0.026a 1.270±0.036a ±0.12ab 10.83±0.41c 71.30±4.40Aa 38.97±3.46ABbcd 0.361±0.014a 1.280±0.030ab ±0.07b 10.00±0.63b 77.82±5.41ABCb 42.07±3.58Bd 0.420±0.013b 1.282±0.028ab ±0.13b ±0.52c 79.05±3.35ABCbc 34.64±3.22Aab 0.377±0.032a 1.317±0.036b ±0.23b 11.00±0.005c 74.62±2.28ABab 38.31±4.07ABabcd 0.356±0.001a 1.297±0.031ab ±0.05a 11.00±0.006c 77.20±2.80ABCb 33.71±1.35Aa 0.368±0.002a 1.307±0.028ab ±0.21ab 11.00±0.006c 79.40±5.86ABCbc 39.82±2.28ABcd 0.377±0.036a 1.283±0.031ab ±0.32ab 11.00±0.003c 84.45±1.77Cc 36.99±5.98ABabc 0.353±0.003a 1.312±0.024ab ±2.10ab 11.00±0.007c 83.56±3.56Cc 36.92±4.32ABabc 0.350±0.013a 1.305±0.035ab Control 60.44±1.01a 9.17±0.408a 77.45±7.94ABCb 36.48±3.21ABabc 0.360±0.021a 1.288±0.021ab Means with different uppercase letters in each row differ with high significance (<0.01); Lower case letters indicate significant ( <0.05) difference Table 4: A regression orthogonal design analysis of variance on egg quality index Z0 Z1 Z2 Z1Z2 Z3 Z1Z3 Z2Z3 Z1Z2Z3 Egg weight bj Egg yolk color bj Hartz unit bj Eggshell strength bj Eggshell thickness bj Egg-shaped index bj bj represents a return to form individual regression coefficient for the corresponding item indicates a significant level of the corresponding factors. could significantly increase the iron content in iron deficiency anemiarat tissue. Further research was needed to clarify on laying hens The effects of rchmannia glutnosa polysaccharides on egg quality and biochemical indicators had rarely been reported. In this experiment, the average egg weight, yolk color, and Hartz unit were all enhanced and improved following the addition of rchmannia glutnosa polysaccharides. The average egg weight was significantly increased (<0.05), as was eggshell strength (<0.01). Triglycerides had a tendency to decrease (<0.1) with the increase of rchmannia glutnosa polysaccharides. There was still very limited data regarding rchmannia glutnosa polysaccharides improving egg quality and serum biochemical indicators; therefore, further study was needed. Sun et al. (2012) and colleagues added 0.5%, 1.0%, 1.5% motherwort powder to the diet of egg-laying hens. And laying hens could only pigment from the diet Instead of itself synthesis. Du et al. ( 2007) found that there were no significant differences in the color of yolk between chmannia glutnosa polysaccharides extracts. This suggested that the egg yellow level was not in cr ea sed wi th a n increase in proport ion of

4 4 INDIAN JOURNAL OF ANIMAL RESEARCH Table 5: Effect of Spirulina on Biochemical Indicators of egg-laying hens (Unit mmol/l) No. Cholesterol Triglycerides Iron Total protein Calcium ±1.22ABa 12.83±0.88BCc 26.59±2.12a 52.47±2.03a 5.85±0.72ABbc ±0.67ABab 9.36±3.27ABCab 21.89±0.97a 54.29±1.48a 4.25±1.36Aa ±1.52ABa 9.56±1.76ABCab 22.01±3.38a 56.72±2.11a 5.30±2.98ABabc ±0.35ABab 11.56±1.59ABCbc 22.08±0.97a 60.82±1.12a 5.324±1.48ABabc ±0.78Aa 9.03±13.59ABab 24.09±1.46a 57.99±4.17a 5.17±1.54ABabc ±0.56ABab 8.63±1.95ABab 24.37±2.01a 53.81±1.72a 4.70±0.63ABab ±0.74ABa 11.36±2.59ABCbc 27.57±1.64a 57.80±2.84a 5.91±2.33ABbc ±0.29Aa 7.74±2.11Aa 25.64±2.06a 52.97±1.72a 4.21±1.55Aa ±2.91ABab 9.04±1.19ABab 23.17±1.49a 58.27±4.29a 6.22±9.14Cc ±2.38ABab 8.97±1.23ABab 23.41±0.57a 57.96±4.66a 6.21±1.43Cc Control 6.47±1.91Bc 13.86± Cc 27.17±1.05a 58.50±0.95a 5.47±1.08ABabc Means with different uppercase letters in each row differ at highly significant levels (<0.01); Lower case letters indicate significant differences ( <0.05) Table 6: A regression orthogonal design analysis of variance on Biochemical Indicators of egg-laying hens Index Z0 Z1 Z2 Z1Z2 Z3 Z1Z3 Z2Z3 Z1Z2Z3 Cholesterol Bj Triglycerides bj Iron bj Total protein bj Calcium bj bj represents a return to form individual regression coefficients for the corresponding item; indicates s a significant level of the corresponding factors. rchmannia glutnosa polysaccharides extracts. Yuan et al. (1998). Eggshell calcium was obtained in large part from the feed (Du et al., 1999) and the main component of rchmannia glutnosa polysaccharides extracts was flavonoids, which influenced calcium metabolism, thus changing the eggshell strength (Liang et al., 2006). When average egg weight and eggshell strength were used as indicators, spirulina powder and rchmannia glutnosa polysaccharides extracts were significantly antagonistic ( <0.01); each additive was especially effective at high levels (spirulina powder is 15g/kg or rchmannia glutnosa polysaccharides extract is 3.5 g/kg). Leonurus japonicus and rchmannia glutnosa polysaccharides are especially antagonistic to eggshell strength and cholesterol levels, (<0.05), especially at high levels (rchmannia glutnosa polysacchar ides extr act a t 3.5 g/kg or Leonurus japonicus at 1.1g/kg). CONCLUSION The results demonstrated that supplementing the diets of egg-laying hens with Spirulina improved egg quality and reduced the cholesterol content of the hens blood. The results of this study show that when spirulina powder is present at a concentration of 10g/kg, rchmannia glutnosa polysaccharides was present at 3g/kg, and Leonurus japonicus was present at 0.9 g/kg, the best results were seen in the Hartz unit,which increased by 9.04% compared to the control group. When spirulina powder was added at 5 g/kg, rehmannia polysaccharide was added at 3.5 g/kg, and Leonurus Japonicas is added at 1.1 g/kg the eggshell strength and eggshell thickness are the largest. Cholesterol and triglycerides were decreased the most when spirulina powder was added at 15g/kg, rchmannia glutnosa polysaccharides was added at 3.5 g/kg, and Leonurus Japonicas was added at 1.1 g/kg. REFERENCES Chan, W. C., Wong Y. C., Kong Y. C., et al. (1983). Clinical observation on the utero tonic effect of l-mu Ts ao ( Leonurus artemisia). Am J Chin Med, 11: Cheng, Z. G. (2001). Broiler skin and egg yolk coloring. Feed wide-angle, 5: Chicken breed standard. (2004). The Ministry of Agriculture of the eople s Republic of China. Dai, R. G., Gao S. J., Deng X. Q. (2012). Feeding effects of spirulina. Jiangxi Agri Sci, 38. Du, L. X., Tang J. Laying hens Manual. Chin Agri Univer ress,

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