Egypt. Poult. Sci.

Vol (32) (I): (201-215)

EFFECT OF USING SPIRULINA PLATENSIS ALGAE AS AFEED ADDITIVE FOR POULTRY DIETS: 1- PRODUCTIVE AND REPRODUCTIVE PERFORMANCES of LOCAL LAYING HENS. By
Mariey, Y. A.;H.R.Samak, and M.A.Ibrahem Animal Production Res.,Instit. Agric.Res. Center, Minist. Of Agriculture. Received: 9/2/2012 Accepted: 9/4/2012

ABSTRACT: The present study was designed to evaluate the productive and reproductive performance of two local strains of laying hens fed Spirulina platensis algae containing diets. An 24 factorial experiment was conducted, two local strains of laying hens [Sinai (S) and Gimmizah (G)] were fed on experimental diet containing 4 levels of Spirulinapowder (0, 0.10, 0.15 or 0.20%) from 28 to 52 weeks of age. A total number of 240 Sinai (S) and Gimmizah (G) pullets (120 of each strain) were randomly divided into four dietary treatments (3 replicates of 10 birds per treatment) and housed in floor pens were used. The basal diet was formulated to contain 2743 kcal/kg metabolizable energy and 16.4% crude protein. The obtained results can be summarizes as follows: Irrespective of hen strain, results showed that birds fed Spirulina-diets achieved superior significantly means of egg production rate, daily egg mass and feed conversion ratio to those of the control group. Analysis of variance also shows that birds fed Spirulina-diets laid significantly heavier eggs than those of control counterparts, regardless of type of hen strain. Feeding Spirulinadiets gave significant increases in egg yolk percentage and yolk color score compared with those of the control group. On the other hand, no significant differences were observed in percentages of egg shell and albumen or Haugh unit due to experimental diets. Also, there were significant (P<0.05) reduction in yolk cholesterol as the level of dietary Spirulina increased. The same response was observed for levels of plasma cholesterol. Data showed no significant differences among different dietary treatments in egg weight loss percentages or chicks weight at hatch, while the fertility and hatchability percentages of eggs produced by birds fed the Spirulina-containing diets were significantly superior compared to those of the control group. Panel test; using fresh or stored eggs, demonstrated that odor intensity, flavor and taste of normal egg (fresh or stored) were insignificantly affected by feeding the different dietary treatments. In the present study, regardless of the effect of dietary inclusion of Spirulina, Gimmizah hens consume significantly more feed than that of Sinai hens, while Sinai birds give significantly higher egg production performance compared with Gimmizah hens. Also, Sinai hens had significantly (P<0.05) a better value of feed conversion ratio than that of Gimmizah hens. In conclusion, taking the economical aspect into account, Spirulina algae could be safely used in laying hen diets with superior effects on their productive and reproductive performance

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INTRODUCTION The recent trend in the feed business is currently directed toward the use of natural ingredients as alternatives to antibiotics, synthetic colors, and other chemicals. Spirulina (blue-green alga) is one of the high quality natural feed additives that can be used in animal and poultry nutrition. There are two different species of Spirulina: Spirulina maxima and Spirulina platensis, with varying distribution throughout the world (Oliveira et al., 1999). Spirulina platensis is more widely distributed and found mainly in Africa, Asia and South America (Vonshak, 2002). The blue-green algae (Spirulina platensis) have been used for hundreds of years as a food source for humans and animals due to the excellent nutritional profile and high carotenoid content. Spirulina is relatively high in protein with values ranged between 55-65% and includes all of the essential amino acids (Bourges et al., 1971; Anusuya Devi et al., 1981). The available energy content of Spirulina has been determined to be 2.503.29 kcal/gram and its phosphorous availability is 41% (Yoshida and Hoshii, 1980; Blum et al., 1976). Also, Spirulina algae are rich in thiamin, riboflavin, pyridoxine, vitamin B12, vitamin C and carotenoids and have been used throughout the world as a feed component in broiler and layer diets to enhance yolk color and flesh (Colas et al., 1979; Brune, 1982; Ross and Dominy, 1985, 1990). In addition, it is rich in nutrients such as vitamins, amino acids, gamma linoleic acid, phycocyanins, tocopherols, chlorophyll and -carotenes (Abd El-Baky et al., 2003 and Khan et al., 2005). It has also been reported that Spirulina has health benefits in conditions such as diabetes mellitus and arthritis (Parikh et al., 2001; Rasool et al., 2006). It
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has also been shown that Spirulina has immuno-stimulatory effects and antiviral activity (Khan et al., 2005). Spirulina has been shown to enhance immune function, reproduction and increase growth. Less than 1% Spirulina added to chicken diets has been found to significantly enhance the defense systems for increased microbial killing, antigen processing and greater Tcell activity (Qureshi, et al., 1994). In this respect, Ross and Dominy (1990) and Nikodmusz et al (2010) reported that hens fed Spirulina-containing diets achieved superior productive and reproductive performance compared to the control birds. Moreover, Sakaida Takashi, (2003) found that egg yolk color was significantly improved by the addition of Spirulina to laying hen diets. In an early study, Ross et al. (1994) found no adverse effect of dietary Spirulina supplementation on egg production, feed per egg, egg weight, body weight or mortality rate. However, others reported did not recorded any effect on reproductive performance of laying hens (Salazar et al., 1996; Ishimi Sugiyama, 2006; Voltarelli and de Mello, 2008). In another study, dietary Spirulina inclusion improved egg fertility, from 87% to over 96% (Ross and Dominy, 1990). Recently, Raju et al. (2005) concluded that dietary inclusion of Spirulina at a level of 0.05% can partially offset the adverse effects of 300 ppm aflatoxin on growth rate and lymphoid organ weight of broiler chickens. More recently, Islam et al. (2009) found that Spirulina may be helpful for reducing the tissue burden of arsenic in ducks. Aim of this study was to investigate the effects of dietary supplementation of Spirulina platensis on productive and reproductive performances of two local strains of laying hens.

Spirulina platensis algae afeed additive local laying hens. MATERIALS AND METHODS This study was carried out at ElGimmizah Poultry Station, Animal Production Research Institute, Agricultural Research Center, Ministry of Agriculture, Egypt during the period from 28 to 52 weeks of age. An 24 factorial experiment which contain two local strains of laying hens [Sinai (S) and Gimmizah (G)] were fed on experimental diet contained 4 levels of Spirulina powder (0, 0.10, 0.15 or 0.20%) during the experimental period from 28 to 52 weeks of age. A total number of two hundred and fourty pullets (120 of each strain) were randomly divided into four dietary treatments (3 replicates of 10 birds per each) and housed in floor pens. The basal diet was formulated to contain metabolizable energy, 2743 kcal/kg; crude protein, 16.4%; calcium, 3.29%; nonphytate phosphorus, 0.344%; lysine, 0.914%; methionine, 0.364% and methionine plus cystine, 0.639% ), and was used to serve as a control diet for the local strains of laying hens. The composition and nutrive value of the experimental diets are presented in Table 1, according to NRC (1994). All birds were kept under the local conditions and fresh water was supplied all the time and all hens were fed ad libitum. Also, the birds were exposed to 16 hr of continuous light. The productive performance of pullets was evaluated as change in body weight, egg production rate, egg weight, daily egg mass, daily feed intake and feed conversion ratio throughout 28-day period basis. At 36 weeks of age, egg quality test was performed to examine certain traits, including egg shape index, egg components (relative weights of shell, yolk and albumen), yolk index, Haugh units (Haugh, 1937) and yolk color score by using the Roche yolk color fan. Egg yolk cholesterol
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was also determined according to the procedures of Elkin and Rogler (1990). Blood samples were collected from five birds per treatment into heparinized tubes and plasma was separated by centrifugation at 3000 rpm for 15 minutes. In blood plasma, concentrations of total lipids (Frings and Dunn, 1970), cholesterol (Allain et al., 1974) and triglycerides (Fossati and Prencipe, 1982) were determined using commercial kits. Concentration of plasma total proteins, albumin and glucose were colorimetrically estimated (Washburn and Nix, 1974). While, plasma globulin concentration was obtained by subtracting the concentration of albumin from total proteins. At 36 weeks of age, all settable eggs per treatment were collected and incubated. Fertility percentage was estimated as a percentage of fertile eggs to the total eggs set. Hatchability of fertile eggs and egg weight loss were also estimated. Individual weights of hatched chicks were recorded. At the end of study (52 weeks of age), a panel test was carried out on boiled fresh eggs or eggs stored for 14 days at 5oC. On days 0 and 14 of storage, ten eggs were randomly selected from each treatment and cooked for the evaluation of sensory characteristics according to the method of Caston et al. (1994). The parameters examined were the intensity of egg odor, flavor, perception of normal or off-flavor, taste and yolk color. The economical efficiency of the tested rations was calculated from input output analysis, assuming that the other costs were constant. A completely randomized design, with a factorial arrangement of treatments (24) was used. Data were analyzed using the General Linear Model procedure of Statistical Analysis System (SAS, 1997). Significant differences among means were separated by Duncan's multiple range test (Duncan, 1955).

Y. A. Mariey, et al.

RESULTS AND DISCUSSION Growth performance changes: Data presented in Table 2 showed that the effect of treatments on initial and final weight of pullets was not significant. However, body weight gain of laying hens fed Spirulina diets increased insignificantly as compared to that of the control group, but the differences were significant (P<0.05) only between hens fed 0.20% Spirulina and the control hens. These results are in agreement with those obtained by Ross et al. (1994), who found that there was no adverse effect of dietary Spirulina on final body weight. However, Ross and Dominy (1990) and Nikodmusz et al. (2010) reported that birds fed dietary Spirulina had benefit effects on productive performance. The significant (P<0.05) effect of dietary treatment on weight gain of hens fed 0.20% Spirulina diet may be brought about through improving the efficiency of feed utilization. In this regard, Raju et al. (2005) concluded that dietary inclusion of Spirulina at a level of 0.05% can partially offset the adverse effects of aflatoxin on growth rate of broiler chickens. Irrespective of dietary inclusion of Spirulina, final body weight and weight gain were significantly (P<0.05) higher in Gimmizah than those of Sinai hens (Table 2). Laying performance parameters: Data presented in Table 3 showed that hen performance in term of egg production, egg weight, egg mass and feed conversion ratio was affected by dietary treatment. All previous traits were significantly (P<0.05) improved by dietary treatment, in particular, with the highest level of Spirulina (0.2%). It is of interest to note that such improvement was associated with insignificant differences in daily feed intake of hens. In line with the present results, several authors reported that laying hens fed Spirulina-containing diets,
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especially those fed 0.2% Spirulinacontaining diet attained the best means of egg production and feed conversion compared with those of the control group (Ross and Dominy, 1990; Ross et al., 1994; Nikodmusz et al., 2010). The increase in egg weight for hens fed the Spirulina-diets may be attributed to heavier egg yolks as indicated in Table 4. In this regard, Ross et al. (1994) found that egg weight was significantly higher for hens fed diets containing different levels of Spirulina. In addition, Sakaida Takashi (2003) and Nikodmusz et al. (2010) observed that feeding Spirulina-containing diets gave a beneficial effect on productive performance of laying hens. Concerning feed conversion ratio, it was significantly (P<0.05) better for pullets fed the Spirulina inclusion, being the best for birds fed 0.2% Spirulina (3.46 g:g), while the worst value (4.54 g:g) was recorded for the control group. Since no differences were observed in feed intake of hens fed the experimental diets containing Spirulina, the significant improvement in feed conversion ratio achieved by hens fed such diets may be due, at least in part, to an improvement in daily egg mass. These results agreed with the findings of several investigators, who reported that addition of Spirulina to the diet improved feed conversion of laying hens (Ross and Dominy, 1990; Ross et al., 1994; Nikodmusz et al., 2010). On the other hand, the improved egg production and daily egg mass of hens fed Spirulina diets could also be contributing factors for the improvement of productive performance achieved by these hens. The present results were supported by the findings of Qureshi et al. (1994), who found that Spirulina could enhance immune function, reproduction and increase growth of broiler chickens. Also, it has been reported that adding less than 1% Spirulina to chicken diets significantly enhanced the

Spirulina platensis algae afeed additive local laying hens. defense systems for increased microbial killing, antigen processing and greater Tcell activity Irrespective of the effect of dietary inclusion of Spirulina, Gimmizah hens consume significantly more feed (118.3 g/hen) than that of Sinai hens (108 g/hen). Meanwhile, Sinai birds significantly yielded (P<0.05) higher egg production performance and significantly (P<0.05) better feed conversion ratio than those of Gimmizah hens (Table 3). Egg quality traits: Results in Table 4 showed that feeding Spirulina-diets resulted in significant (P<0.05) increase in egg yolk percentage and yolk color score compared with those of the control group. While, the effect of dietary treatment on percentages of egg shell and albumen or Haugh unit was not significant. These results are in line with those obtained by Inborr (1998), who reported that there were no significant differences in egg shell percentage, yolk index, albumen percentage and Haugh unit as result of feeding experimental diets containing Spirulina. Also, it was reported that feeding Spirulina-diets achieved a significant increase in yolk color score (Sakaida Takashi, 2003) and egg yolk color score from quails fed dried spirulina (Ross et al., 1994). Generally, a linear relationship between dietary Spirulina concentration and egg yolk pigmentation was recorded by Anderson et al. (1991) on Leghorn hens. Egg fertility and hatchability: Data presented in Table (5) show that the effect of dietary treatment on egg weight loss percentage or chicks weight at hatch was not significant. However, percentages of fertility and hatchability of eggs produced by birds fed the Spirulina diets were significantly (P<0.05) higher than those of the control group. Such increase may be related to the high contents of tocopherols containing Spirulina. In this
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respect, El-Khimsawy (1985) reported that tocopherols had a vital role in fertility and hatchability of poultry. The current results are in line with those obtained by Ross and Dominy (1990), who reported that Spirulina inclusion in hen diets resulted in an improvement in egg fertility, from 87% to over 96%. Also, Inborr (1998) reported that the improvement of egg fertility for broiler breeders was recorded when Spirulina incorporated into their diets reflected a 5% improvement in hatchability rates. Results showed no significant differences between Sinai and Gimmizah hens in all previous traits, with the exception of hatch weight of chicks, being significantly (P<0.05) higher for Sinai hens than that of Gimmizah hens. Blood parameters: Blood plasma concentrations of cholesterol, triglycerides and total lipids were significantly (P<0.05) lower in birds fed diets containing Spirulina (Table 6). The lowest concentration of studied blood biochemicals were recorded for hens fed 0.2% Spirulina-diet, followed by those fed 0.15 and 0.1% Spirulina-diets, respectively. In line with the present results, Torres et al. (1998) and Fong et al. (2000) reported a significant reduction of triglycerides and cholesterol concentrations for rats or mice fed Spirulina diets. Regarding the strain differences, results in Table 6 showed that Gimmizah hens exhibited lower concentration of plasma cholesterol, triglycerides and total lipids levels than those of Sinai hens. Yolk total lipids and cholesterol: There were significant (P<0.05) reductions in yolk cholesterol and total lipids as the level of dietary Spirulina was increased (Table 6). The reduction rate of yolk cholesterol relative to that of the control group exhibited by birds fed the 0.1, 0.15 and 0.2% Spirulina-diets was

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13.0, 19.8 and 24.4%, respectively. The corresponding rates of yolk total lipids were 6.7, 16.6 and 21%. This reduction in yolk total lipids and cholesterol contents may be related to their lower levels in blood plasma of hens fed the Spirulina-containing diets. The current results are in harmony with those reported by Sakaida Takashi (2003), who found that hens fed tested diets supplemented with varying levels of Spirulina tended to decrease the cholesterol content in the egg yolk of hens. Plasma proteins and glucose: Effects of dietary Spirulina supplementation on concentration of plasma total proteins, albumin and globulin are presented in Table 7. Feeding dietary Spirulina had a significant (P<0.05) increase in the levels of plasma total protein, albumin and globulin as Spirulina levels in diets increased up to 0.2%. The same response was observed for plasma glucose concentration of hens fed dietary Spirulina. The increased concentrations of plasma total protein, albumin and globulin may be related to the high protein contents in Spirulina (with values ranging from 5565% and includes all of the essential amino acids). These results are in line with the findings of Tewe (1985) and Eggum (1989), who stated that total serum protein, globulin and albumin were directly responsive to both protein quantity and quality. The increase in plasma glucose concentration of hens fed dietary Spirulina may be attributed to its excellent nutritional profile and high carotenoid content. In this regard, El-Khimsawy (1985) reported that vitamin A plays an important role for synthesis glucose molecule in the body. There were no significant differences in yolk cholesterol and total lipids contents between the two strains, regardless of the effect of dietary inclusion of Spirulina (Table 6).
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Sensory evaluation: The effect of feeding dietary Spirulina on sensory quality of fresh and stored eggs is presented in Table 8. Data of the panel test using fresh or stored eggs demonstrated that odor intensity, flavor and taste of normal egg (fresh or stored) were insignificantly affected by feeding the different dietary treatments, irrespective of hen strain. On the other hand, the data in the present study showed that feeding dietary Spirulina resulted in significant increases in yolk color score of normal eggs (fresh or stored) compared with those of the control group. From these results it can be speculated that the higher yolk color values in the case of raw eggs might be due to the brightness of the vitelline membrane and/or higher level of pigment deposition at the periphery rather than towards the central region of the yolks. These results are in line with those obtained by Saxena et al. (1982), who reported that the organoleptic parameters, examined herein, revealed that eggs from hens fed Spirulina had significantly higher scores than those of the control hens. Data in Table 8 showed that the panelists were not able to distinguish between eggs from Sinai and Gimmizah hens that fed dietary Spirulina. Economic efficiency: Results presented in Table 9 showed that economic efficiency of birds fed dietary Spirulina was superior to that of the control group. Birds fed the 0.2% Spirulina had the highest economic efficiency (63.8%), while the lowest one (31.4%) was recorded for the birds fed the unsupplemented control diet (0.0% Spirulina). Also, economic efficiency of Sinai hens was higher than that of the Gimmizah hens.

Spirulina platensis algae afeed additive local laying hens. Generally, the best results of the productive and reproductive performance or egg quality in the present study may be related to one or more of the following points: 1- Spirulina has an excellent nutritional profile (high carotenoids, high protein with includes all of the essential amino acids and rich in minerals and vitamins (Bourges et al., 1971; Anusuya Devi et al., 1981; Brune, 1982; Ross and Dominy, 1985, 1990). 2- Spirulina has been shown to enhance immune function, reproduction and growth, as reported by Quereshi et al. (1994) and Khan et al. (2005). 3- Feeding Spirulina containing diets may increase the lactobacillus population and enhance the absorbability of dietary vitamins (Tokai et al., 1997). In conclusion, taking the economical aspect into account, Spirulina algae could be safely used in laying hen diets, at level of 0.2%, with superior effects on their productive and reproductive performance. Table 1: Composition and chemical analyses of the basal diets.
Ingredient Yellow corn Soybean meal, 44 % Limestone Dicalcium phosphate Common salt (NaCl) Vit. & Min. mix.* Methionine Total Calculated values**: Crude protein, % ME,Kcal/kg Crude fiber,% Ether Extract,% Calcium, % Available phosphorus, AP % Total phosphorus % Lysine, % Methionine,% Methionine + cysteine % Determined values***: Dry matter, % Crude protein,% Crude fiber,% Ether Extract,% NFE% Ash,% (%) 67.90 23.00 7.00 1.50 0.30 0.25 0.05 100 16.119 2787.675 3.102 2.762 3.300 0.405 0.671 0.794 0.314 0.588 90.112 15.815 3.859 2.511 58.359 9.568

*Vit.& Min. mix:. each 3kg contains: 10,000,000 IU Vit. A; 2,000,000 IU Vit D3 10,000 mg Vit. E;1,000mg Vit. K; 1,000mg Vit. B1; 5,000mg Vit. B2; 1,500mg Vit B6; 10mg Vit. B12; 30mg; Niacin, 20 gm ; Panatothenic acid, 1gm, Biotin;I,000mg Folic acid;250,000mg choline chloride; 80gm manganese; 40gm iron; 40gm zinc; 2gm copper; 2gm iodine; 1gm Seleinium and 1gm cobalt. ** Calculated according to NRC (1994). *** Determined according to the methods of A.O.A.C (1980) 207

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Table 2 : Main effects and standard errors of initial, finial live body weight and weight gain of laying hens as affected by dietary treatment and strain of hens. Initial body (g/hen) Effect of Spirulina level (%): Control 1352.7 0.10 1328.4 0.15 1327.7 0.20 1339.5 SEM1 17.354 Significance NS Effect of strain: Sinia 1310.7b Gimmizah 1363.5a SEM1 12.271 Significance * Main effects weight Finial body weight Total gain (g/hen) (g/hen) 1470.5 1455.9 1453.0 1471.0 18.510 NS 1427.7b 1497.5a 13.089 * 117.8b 127.5ab 125.3ab 131.5a 3.966 * 117.0b 134.0a 2.804 *

a-b :Means with different superscripts within the same column for each variable are significantly different at P0.05. NS: not significant; *: significant at P0.05. 1: Standard error of the means. Table 3: Main effects and standard errors of egg production, egg weight, egg mass, feed intake and feed conversion as affected by dietary treatment and strain of hens. Egg production (%) Effect of Spirulina level (%): Control 52.3b 0.10 60.3a 0.15 61.8a 0.20 63.3a SEM1 0.010 Significance * Effect of strain: Sinai 63.0a Gimmizah 55.8b SEM1 0.007 Significance * Main effects Egg weight (g) 48.05c 49.48b 51.55a 51.82a 0.373 * 49.22b 51.18a 0.264 * Daily egg Daily feed Feed mass intake conversion (g/hen) (g/hen) ratio (g:g) 25.11c 29.72b 31.83a 32.69a 0.620 * 31.06a 28.61b 0.444 * 113.7 113.5 113.0 112.5 0.825 NS 108.0b 118.3a 0.583 * 4.54a 3.84b 3.57c 3.46c 0.069 * 3.52b 4.18a 0.052 *

a-c :Means with different superscripts within the same column for each variable are significantly different at P0.05. NS: not significant; *: significant at P0.05. 1: Standard error of the means.

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Spirulina platensis algae afeed additive local laying hens. Table 4: Main effects and standard errors of egg quality of laying hens as affected by dietary treatment and strain of hens. Shell Yolk weight weight (%) (%( Effect of Spirulina level (%): Control 12.60 31.10b 0.10 12.96 32.62a 0.15 12.94 32.90a 0.20 12.30 31.82ab SEM1 0.215 0.478 Significance NS * Effect of strain: Sinai 12.84 31.88 Gimmizah 12.56 32.34 SEM1 0.152 0.338 Significance NS NS Main effects Yolk index (%) 39.63 41.74 42.16 42.53 0.006 NS 41.61 41.42 0.004 NS Albumen (%) 56.29 54.41 54.15 55.87 0.467 NS 55.27 55.10 0.330 NS Haugh Units 88.05 89.80 88.40 87.98 1.053 NS 88.64 88.46 0.745 NS Yolk Color 4.8c 6.3b 6.7b 7.6a 0.256 * 6.2 6.5 0.202 NS

a-c :Means with different superscripts within the same column for each variable are significantly different at P0.05. NS: not significant; *: significant at P0.05. 1: Standard error of the means. Table 5: Main effects and standard error of egg weight, egg weight loss, fertility, hatchability and chicks weight of Sinai and Gimmizah laying hens as affected by the different dietary treatments. Egg Main effects weight (g) Effect of Spirulina level (%): Control 52.01 0.10 52.68 0.15 53.07 0.20 53.35 SEM1 0.423 Significance NS Effect of strain: Sinai 52.05 Gimmizah 53.05 SEM1 0.299 Significance NS egg weight loss % 15.57 15.46 15.45 15.34 0.106 NS 15.49 15.42 0.075 NS Egg fertility % 90.87a 94.62b 95.68b 96.58bc 0.396 * 94.90 93.97 0.396 NS Egg hatchability Chicks weight (g) 31.24 31.26 32.13 32.22 0.407 NS 30.94a 32.73b 0.284 *

89.81a 93.17b 95.10b 95.75b 0.967 * 93.97 93.00 0.684 NS

a-c :Means with different superscripts within the same column for each variable are significantly different at P0.05. NS: not significant; *: significant at P0.05. 1: Standard error of the means.

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Table 6 : Main effects and standard error of plasma cholesterol, total lipids and triglyceride of Sinia and Gimmizah laying hens as affected by the different dietary treatments. Blood plasma Main effects Cholesterol mg/100ml Egg yolk Total Total Triglyceride Cholesterol lipids lipids mg/100ml mg/yolk mg/100m mg/g yolk 314.83a 306.33b 298.83c 291.00d 2.146 * 305.58a 299.91b 1.517 * 9.40a 8.93a 8.75b 8.58b 0.078 * 8.88 8.95 0.055 NS 284.2a 265.0b 237.1c 224.6d 3.344 * 252.1 253.3 2.364 NS 13.50a 11.75b 10.83c 10.20c 0.298 * 11.37 11.75 0.210 NS

Effect of Spirulina level (%): Control 116.25a 0.10 112.55ab 0.15 109.96b 0.20 108.91b SEM1 1.341 Significance * Effect of strain: Sinai 112.41 Gimmizah 111.42 SEM1 0.949 Significance NS

a-d:Means with different superscripts within the same column for each variable are significantly different at P0.05. NS: not significant; *: significant at P0.05. 1: Standard error of the means. Table 7: Main effects and standard error of plasma globulin albumin total protein and glucose of Sinia and Gimmizah laying hens as affected by the different dietary treatments. Main effects Globulin g/100ml Albumin g/100ml 2.33a 2.28ab 2.38ab 2.52b 0.123 * 2.35 2.30 0.087 NS Total protein g/100ml 5.91a 6.30a 7.03b 7.50c 0.140 * 6.63 6.74 0.099 NS Glucose mg/100ml 320.33a 326.17ab 333.83bc 339.50c 2.889 * 331.75 328.17 2.043 NS

Effect of Spirulina level (%): Control 3.88a 0.10 4.02ab 0.15 4.52b 0.20 5.12c SEM1 0.159 Significance * Effect of strain: Sinai 4.38 Gimmizah 4.38 SEM1 0.113 Significance NS

a-c :Means with different superscripts within the same column for each variable are significantly different at P0.05. NS: not significant; *: significant at P0.05. 1: Standard error of the means.

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Spirulina platensis algae afeed additive local laying hens. Table 8 : Main effects and standard error of scores of panel test characteristics of eggs produced by Sinai and Gimmizah laying hens as affected by the different dietary treatments. Odor fresh stored Effect of Spirulina level (%): Control 4.50 4.20 0.10 4.55 5.55 0.15 4.95 4.95 0.20 5.2 5.20 SEM1 0.299 0.323 Significance NS NS Effect of strain: Sinai 4.90 4.60 Gimmizah 4.85 5.85 SEM1 0.211 0.228 Significance NS NS Main effects Flavor Taste Yolk color fresh stored fresh stored fresh stored 4.90a 5.10ab 5.65ab 5.75b 0.263 * 5.45 5.35 0.186 NS 4.75 5.10 5.20 5.35 0.273 NS 5.22 4.97 0.193 NS 5.25 5.20 5.40 5.65 0.374 NS 5.30 5.45 0.265 NS 5.10 5.00 4.90 5.45 0.295 NS 4.97 5.25 0.208 NS 5.7a 7.1b 7.8b 7.2b 0.287 * 7.1 7.2 0.203 NS 5.6a 7.0b 7.8b 7.9c 0.265 * 7.0 7.2 0.187 NS

a-c :Means with different superscripts within the same column for each variable are significantly different at P0.05. NS: not significant; *: significant at P0.05. 1: Standard error of the means. Table (9): Economic efficiency of Sinai and Gimmizah laying hens as affected by dietary treatment. Item Feed cost/hen/day (L.E) No. of eggs /hen/day (L.E) Total revenue/hen/day Net revenue/hen/day (L.E) Economic efficiency (%) 0% 0.239 0.523 0.314 0.075 31.4% Spirulina level 0.1% 0.15% 0.238 0.235 0.603 0.618 0.362 0.371 0.127 0.136 52.1% 57.9% Streain 0.2% Sinia Gimmizah 0.232 0.225 0.247 0.633 0.630 0.558 0.380 0.378 0.335 0.148 0.153 .0.088 63.8% 68.0% 35.6%

Feed cost/hen (L.E) = Feed intake /hen/day price of kg feed. Total revenue/hen (L.E) = No. of eggs /hen/day (L.E) price of an egg at time of experiment=0.60 L.E REFERENCES Association of Official Analytical Chemists (AOAC, 1980).Official methods of analysis '13th Ed., published by the AOAC, Benjamin Franklin Station.Washington,.D.C Abd El-Baky, H.H.; El-Baz, F.K.; and ElBaroty, G.S. (2003). Spirulina species as a source of carotenoids
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and -tocopherol and its anticarcinoma factors. Biotechnology, 2: 222-240. Allain,C.A; Poon, L.S.; Chan,C.G.S; Richmons,W. and Fu, P.C. (1974). Enzymatic determination of total serum cholesterol. Clin. Chem., 20:470475.

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