Abstract: The objective of this study was to carry a comparative study of carcasses characteristics and fatty acid composition of intramuscular and subcutaneous fat of Awassi and Najdi sheep. The experiment utilized four Awassi lambs and four Najdi lambs fed with 50% alfalfa hay and 50% concentrate feed for a period of 15 days prior to slaughtering. The muscles of the carcasses were analyzed for moisture, protein, fat and ash contents by following the standard methods. The live body weight, carcass diameter and length, protein and fat percentages were significantly lower for Awassi than Najdi breed. The SFA and MUFA were significantly higher for Najdi and Awassi breeds, respectively. Awassi breed showed a higher percentage of C15:0 in intramuscular fat and C15:1 in Longissimus dorsi, semimembranosus and subcutaneous fat than Najdi. Najdi breed showed a higher percentage of C16:0 in Longissimus dorsi, semitendinosus and subcutaneous fat. The C18:0 was significantly higher for Najdi breed in semimembranosus and semitendinosus whilst the C18:1 (trans-9) was significantly higher in Longissimus dorsi and subcutaneous fat of Awassi carcasses. Najdi breed showed significantly higher proportion of SFA in semimembranosus and semitendinosus while MUFA was significantly higher for Awassi carcasses in these muscles and the n-6/n-3 in Longissimus dorsi. In conclusion, the Awassi breed produced better and healthier meat due to low percentage of SFA and high proportion of MUFA compared to Najdi breed.
INTRODUCTION
Consumption of lamb meat is high in the Kingdom of Saudi Arabia like many countries in the tropic and sub-tropic regions. Studies on the fatty acid composition of lamb meat has increased due to their effects on human health (Hoffman et al., 2003). Therefore, the nutritionists in many countries focused on studying the factors affecting the fatty composition in animal meat to produce healthy meat for human consumption. As a result, an increased interest has occurred in recent years to study the fatty acid composition of meat as a major source of fat in the human diet especially of Saturated Fatty Acid (SFA). In human diet, the amount of SFA is considered as one of the major factors harmful in many diseases. Some studies showed that increasing the SFA in human diets is positively correlated with increasing the incidence of the colorectal cancer (Petrik et al., 2000a), increasing blood cholesterol and coronary heart disease mortality (Grundy, 1994; Santos-Silva et al., 2002).
In contrast, Sinclair and O’dea (1990) reported that human diets rich in some essential fatty acids such as UFA, especially polyunsaturated fatty acid (PUFA) like linoleic (C18:2) and linolenic (C18:3) acids, has beneficial effects on the human health. Hansen Petrik et al. (2000b) suggested the importance of the dietary monounsaturated fatty acids (MUFA) such as C16:1 and C18:1 on human health.
Ruminants such as sheep also naturally produce Conjugated Linoleic Acid (CLA) which may have a range of nutritional benefits in the diet (Enser et al., 2001). There are several forms of CLA but the common type is cis-9-trans-11 that is found up to 80% of the total content of the CLA in beef (Chin et al., 1992). The CLA recognized as a source of anticarcinogenic components (Parodi, 1997). West et al. (1998) reported that CLA resulted in lowering the blood cholesterol level. Ip et al. (1994) showed that CLA reduced the incidence of coronary heart disease, decreased food intake, acted as anti-cancer, prevent hardening of the arteries, lowered the amount of the stored body fatty acids and supported the immune system. In term of human nutrition, trans CLA has an undesirable effect not only on plasma low density lipoprotein and high-density lipoprotein but also on coronary heart disease mortality (British Department of Health, 1994).
Recently, the UFA:SFA ratio in human diet is one of the indexes used for nutritional values of fat (Santos-Silva et al., 2002). According to the recommendation of British Department of Health (1994), the intake of SFA in the human diet should be reduced and that of PUFA be increased. In general, identifying the percentages of the different SFA and USFA in the sheep meat help in the development of systems of grading and marketing based on meat appropriate parts, recognize the best breeds, helping the consumer to select the meat parts based on lowest and highest percentage of SFA and USFA, respectively for health reasons.
People of Saudi Arabia and other region countries prefer to slaughter low weight young animals due to better quality meat (Bernabeu and Tendero, 2005). However, in term of production, the producers may benefit from heavier carcasses weight due to increasing their profit. Therefore, production of carcasses meat that satisfy both the consumers and producers with respect to fatty acid composition and profit is the main target.
It is well known that sheep breed is one of many factors affecting the fatty acid composition of intramuscular and subcutaneous fats of the sheep meat (Yoshimura and Namikawa, 1983; Eichhorn et al., 1986). However, very few studies have been conducted to evaluate the effect of sheep breed on the fatty acid composition. Therefore, the main objective of the present study was to compare the fatty acids composition of Longissimus dorsi, Semimembranous, Semitendinos and Subcutaneous fat in two sheep breeds named Awassi and Najdi that are well accepted and liked by Saudi people.
MATERIALS AND METHODS
Animals and sampling: The study was carried at the Agricultural Experiment Station of King Faisal University, the Kingdom of Saudi Arabia. Four Awassi lambs and four Najdi lambs were purchased from a local market and fed 50% alfalfa hay and 50% concentrate feed for a period of 15 days prior to slaughtering to ensure that all the lambs were fed on the same diet. The ingredients of the concentrate feed were approximately 60% barley and 40% wheat bran including a source of vitamins and minerals. All the lambs were individually fed in metabolic crates with free access to water. Animals were treated for internal and external parasites. Then, each lamb was weighed and slaughtered at a slaughterhouse in Al-Ahsa at the same time. Then the carcass weight, length and diameter were recorded immediately after slaughtering. The carcasses were hung by the hind legs and cut into two longitudinal halves (left and right halves). The muscles in the left half of the carcasses were analyzed to determine the moisture, protein, fat and ash contents using the standard methods (No. 981.10, 991.36, 950.46 and 920.153, respectively) according to AOAC (2002). The Longissimus dorsi length for each carcass was measured and penetrated via three sites to determine the average value of the Longissimus dorsi pH and temperature directly after slaughtering using a digital pH meter.
Fatty acid analyses: Fat samples of three intramuscular (Longissimus dorsi, semimembranous and semitendinos) and subcutaneous fat between the 9th and 10th rib from each right half of the carcass were separately collected, numbered, vacuum-packaged in polyethylene bags and stored at -25°C until the determination of fatty acid composition. The fatty acid profile was determined by Soxhlet method (Folch et al., 1957; Ulberth and Reich, 1992). The determination of fatty acids profile was performed by gas chromatography with a column temperature of 200°C and the detector temperature at 250°C. The fatty acid contents were presented as percentages of the total fatty acids.
Statistical analysis: Each male lamb was considered as one experimental unit. The data obtained were subjected to one-way analysis of variance (ANOVA) and expressed as means with a standard error of mean (Mean±SEM) using the general linear (GLM) model program SPSS (SPSS 14.0 SPSS Inc. Inc., Chicago, IL, USA). The treatment means were separated (p<0.05) using Duncan's multiple range test, to evaluate the effect of Awassi and Najdi breeds on carcass characteristics and fatty acid composition of intramuscular and subcutaneous fats. In addition to individual fatty acid data, the data were also summed by the type of fatty acid to obtain the n-6/n-3 and PUFA:SFA ratios.
RESULTS AND DISCUSSION
Effect of breed on carcass characteristics: The effects of Awassi and Najdi breeds slaughtered at 12 months of age on the lamb carcass characteristics were shown in Table 1. There were significant differences (p≤0.05) in live body weight, carcass diameter and length, Longissimus dorsi pH, carcass moisture, protein and fat percentages between Awassi and Najdi breeds. However, the differences in carcass weight, Longissimus dorsi length, temperature and carcass ash percentage were not significant (p≥0.05) (Table 1). Live body weight, carcass diameter and length, protein and fat percentages were significantly lower for Awassi than Najdi breed. The differences in carcass diameter and length, moisture, fat and protein percentages were probably due to the significant differences in the live body weight between Awassi and Najdi breeds.
| Table 1: | Carcass characteristics measured for Awassi and Najdi breeds |
| a,bMeans within a row with different superscripts differ (p≥0.05) | |
The differences in the carcass fat percentage between Awassi and Najdi breeds obtained from this study were in agreement with Zembayashi (1993) who reported differences in fat deposition ability among different breeds. Abdullah and Qudsieh (2008) showed that the total fat in Awassi breed increased with an increase in the slaughter weight. The higher percentage of fat in Najdi breed may be attributed to heavier body weight compared to Awassi. In contrast, Longissimus dorsi pH and carcass moisture percentage were higher for Awassi than Najdi. McGeehin et al. (2001) reported significant differences between breeds in the lamb carcass pH which disagree with the observations of Celik and Yilmaz (2010) who did not find any significant differences in the Longissimus dorsi pH post-slaughter between breeds. Caneque et al. (2001) reported that carcass pH post slaughter values were immediately influenced by live weight at the time of slaughtering. This finding was in agreement with the results obtained in this study. In contrast, Teixeira et al. (2005) showed that live weight had no effect on pH after 1 h of slaughtering. Ekiz et al. (2009) mentioned that pre-slaughter handling was one of the reasons for the difference in the lamb carcass pH after slaughtering. In addition, McGeehin et al. (2001) showed that the degree of pH decline in the lamb carcass after slaughtering was influenced by several factors such as sex, season, age, stress, electrical stimulation, cooling, muscle used and the amount of glycogen deposits which are controllable. All the lambs used in this study were males of the same age, fed with similar diet for a period of 15 days prior to slaughtering. The lambs were slaughtered at the same time and same conditions. The pH of the same muscle was measured to minimize the effect of any factor mentioned above affecting the lamb carcass pH after slaughtering rather than breeds.
Effect of breed on fatty acids composition of the whole carcass: The differences in the fatty acid composition between Awassi and Najdi breeds are shown in Table 2. The pentadecylic (C15:0) (C15:1), margaric (C17:0), heptadecenoic (C17:1) and elaidic (C18:1 trans-9) and MUFA were significantly (p≥0.05) higher for Awassi breed. On the other hand, Najdi breed showed significantly high percentage of palmitic (C16:0), stearic (C18:0), linolenic (C18:3) and total SFA than Awassi breed. However, the differences in capric (C10:0), lauric (C12:0), myristic (C14:0), palmitoleic (C16:1), oleic (C18:1), linoleic (C18:2), CLA, arachidic (C20:0), gadoliec (C20:1), total PUFA, the PUFA/SFA and the n-6/n-3 ratios were not significantly different (p≥0.05) among the two breeds. These results agree with the findings of Roberts (1966), Garcia et al. (1979), Eichhorn et al. (1986), Huerta-Leidenz et al. (1996) and Perry et al. (1998) who reported significant differences in the fatty acid composition among different breeds. The fatty acids showed the largest proportion in the carcasses for both breeds and were C14:0, C16:0, C18:0 and C18:1. These findings are in agreement with Al-Suwaiegh and Al-Shathri (2014) in Awassi lamb, Mahgoub et al. (2002) in goat meat muscle and Bures et al. (2006) in Aberdeen angus and Hereford muscles. Awassi breed contained significantly higher percentage of C15:0 and C17:0 while Najdi breed contained significantly high percentage of C16:0 and C18:0. The C16:0 made up the greatest proportion of SFA in both breeds which agree with the findings of Zapletal et al. (2010). The total SFA was significantly (p≥0.05) higher for Najdi compared to Awassi breed. Ashes et al. (1992) observed that heavier and fatty carcasses contain high percentage of SFA than PUFA. This may explain the high percentage of SFA in Najdi with the highest live body weight compared with Awassi breed in this study.
All the SFA do not have cholesterol elevating effect. At present, three types of fatty acids namely lauric acid (C12:0), myristic (C14:0) and palmitic acid (C16:0) are known to raise blood cholesterol level. Therefore, it increases the incidence of heart disease mortality (Grundy, 1994; Kromhout et al., 1995).
| Table 2: | Differences in the fatty acid composition (%) between Awassi and Najdi breeds |
| a,bMeans within a row with different superscripts differ (p≥0.05), SFA: Saturated fatty acids (C10:0+C12:0+C14:0+C15:0+C16:0+C17:0+ C18:0+C20:0), MUFA: Monounsaturated fatty acids (C14:1+C15:1+C16:1+C17:1+C18:1+C20:1), PUFA: Polyunsaturated (C18:2+C18:3), CLA: cis-9, trans-11 conjugated linoleic acid | |
In addition, Yu et al. (1995) suggested that C14:0 is 4-5 times more hyper-cholesterolaemic than either C12:0 or C16:0. Therefore, Awassi meat contained less undesirable proportion of a particular SFA in term of human health. However, Bonanome and Grundy (1988) and Zock and Katan (1992) reported that stearic acid (C18:0) has no effect on raising the blood cholesterol even though it is one of the SFA. Besides, Nicolosi (1997) and Hassel et al. (1997) showed that C18:0 may exert hypo-cholesterolemic effect on blood cholesterol. This contradiction for the effect of C18:0 may explain why this recommendation has not been implemented in dietary guidance or nutrition labeling. Awassi breed produced high percentage of the C15:1, C16:1, C18:1 and C18:1 (trans-9) than Najdi breed. These results were similar with the findings of Webb and Casey (1995) who reported significant differences in the C18:1 fatty acid among sheep breeds. Meat from Awassi contained significantly high percentage of MUFA than Najdi breed. This result do not agree with the finding of Huerta-Leidenz et al. (1996) who showed that MUFA in Herford and Brahman bulls increased with the highest live weights. While, Orellana et al. (2009) did not find any correlation between the live weight and MUFA. Sturdivant et al. (1992) postulated that C18:0 is desaturated by stearoyl-CoA desaturase to their corresponding C18:1 (n-9) in some breeds. Therefore, this study results suggest that Awassi breed might has a genetic ability to produce carcass fats containing high concentrations of MUFA such as the C18:1 (n-9) than Najdi breed through increasing activity of stearoyl-CoA desaturase. MUFA are beneficial because they increase esterification of cholesterol in the liver. Therefore, when MUFA are used in prescribed amounts, these reduce free cholesterol pool and increase receptor-mediated uptake of low density lipoprotein cholesterol, resulting in a decrease in blood cholesterol levels as reported by the Dietary Guidelines Advisory Committee (DGAC) on the Dietary Guidelines for Americans in 2010. Some studies showed that oleic acid (C18:1) does not raise the level of blood cholesterol (Denke, 1994). The concentration of C18:2 in both the Awassi and Najdi breeds was not significantly different. These results do not agree with the finding of Matsushita et al. (2010) who showed that the C18:2 was high in lambs with les slaughter weight. The results from this study showed that C18:3 increased with increasing the weight at slaughtering and are similar to the findings of Huerta-Leidenz et al. (1996). Recently, nutritionists have focused on the ratio between n-6 and PUFA formed from linoleic acid (C18:2 n-6) and n-3 and PUFA formed from linolenic (C18:3 n-3) in the diet (Williams, 2000). In this study, the n-6/n-3 ratio was not significantly different between the two breeds. The lamb carcass meat from both the breeds offered n-6/n-3 ratio which is above the maximum recommended value for healthy diet (4.0) according to the British Department of Health (1994). Celik and Yilmaz (2010) reported that n-6/n-3 ratio should be low in healthy diet. While, Scollan et al. (2006) reported that the values of n-6/n-3 ratio less than 4 are considered as the most favorable to prevent some cardiovascular diseases. The PUFA/SFA ratio is an important indicator for the negative effects of SFA on blood cholesterol. The minimum PUFA/SFA ratio set for human nutrition is 0.45 (Simopoulos, 2004) and generally it should be around 0.7 (Raes et al., 2003). However, this ratio may not be considered an adequate way to assess the nutritional value of fat because all the SFA correlate positively with cardiovascular diseases and ignore the effects of MUFA (Santos-Silva et al., 2002). The ratio of PUFA/SFA in this study was not significantly different between the two breeds and was less than the recommended value. The results obtained from this study are in agreement with the finding of Celik and Yilmaz (2010) who showed that the PUFA/SFA ratio for Awassi was 0.14.
Effect of breed on fatty acid composition of intramuscular and subcutaneous fats: The effect of breed on fatty acid composition of three intramuscular (Longissimus, semimembranosus and semitendinos) and subcutaneous fat of Awassi and Najdi lamb carcasses is presented in Table 3. Awassi and Najdi carcasses showed no significant difference (p≥0.05) in term of C10:0 and C12:0. The C14:0 was significantly (p≥0.05) high in the semimembranosus for Najdi breed without significant difference in other muscles and subcutaneous fat. Intramuscular fat of Awassi carcasses contained significantly high percentage of C15:0 than Najdi without significant difference in term of subcutaneous fat. The C16:0 was significantly (p≥0.05) high in Najdi carcasses in Longissimus dorsi, semitendinosus and subcutaneous fat and C18:0 in semimembranosus and semitendinosus whilst the C17:0 was significantly (p≥0.05) high in intramuscular and subcutaneous fat for Awassi carcasses. This result agree with the findings of Matsushita et al. (2010) who showed that C16:0 was higher in Longissimus dorsi of heavier lambs compared to lambs slaughtered at lighter weight but disagreed with Borys et al. (2012) who found that the percentage of C17:0 was high in intramuscular fat of heavier lamb carcasses. However, the result in this study do not agree with the conclusions of Caneque et al. (2005) who showed that the proportion of C18:0 was low in the heavier than the lighter carcasses in Longissimus dorsi of Manchego suckling lambs. The total SFA was significantly higher in the Najdi carcasses in semimembranosus and semitendinosus which may be attributed to higher body weight of Najdi breed. The high level of saturation may affect the flavor of meat because SFAs, which solidify very fast compared to UFAs when cold, affect palatability of the meat (Banskalieva et al., 2000). The C14:0, C16:0 and C18:0 represented the highest percentage of SFA in intramuscular and subcutaneous fat for both breeds and C16:0 represented more than a half of the SFA.
| Table 3: | Comparison of fatty acid contents (%) in the intramuscular and subcutaneous fats |
| a,bMeans within a row with different superscripts differ (p≥0.05), SFA: Saturated fatty acids (C10:0+C12:0+C14:0+C15:0+C16:0+C17:0+ C18:0+C20:0), MUFA: Monounsaturated fatty acids (C14:1+C15:1+C16:1+C17:1+C18:1+C20:1), PUFA: Polyunsaturated (C18:2+C18:3), CLA: cis- 9, trans-11 conjugated linoleic acid | |
High values of these SFA are due to the fact that most unsaturated fatty acids are biohydrogenated to saturated fatty acids because of the rumen microorganisms, which may convert PUFA to SFA (Wood and Enser, 1997). This result is similar to that found by De Freitas et al. (2014) who showed that intramuscular fat of Longissimus dorsi of three steer breeds contains the highest percentage of these fatty acids and Bonvillani et al. (2010) who reported that the major SFA in intramuscular fat of goats are C14:0, C16:0 and C18:0. However, Caneque et al. (2005) indicated that C14:0, C16:0 and C18:0 showed the highest proportion of SFA in intramuscular and subcutaneous fat of suckling Manchego lambs. It is clear that the C14:0, C16:0 and C18:0 represent the highest percentage of SFA in all ruminant and monogastric species. The C15:1 and C17:1 were significantly (p≥0.05) higher for Awassi carcasses in Longissimus dorsi, semimembranosus and subcutaneous fat while no significant difference (p≥0.05) was found in semitendinosus. Awassi breed had higher percentage of C18:1 (trans-9) in Longissimus dorsi and subcutaneous fat. In semimembranosus and semitendinosus muscles, although the Awassi breed was lower in live weight compared to Najdi breed, Awassi carcasses displayed higher proportion of MUFA compared to Najdi carcasses without significant difference in Longissimus and subcutaneous fat. This result agrees with the data reported by Oka et al. (2002) who showed that percentage of MUFA in intramuscular fat in steers was negatively correlated with body weight. Beef flavor has been shown to be positively correlated with the percentage of MUFA (Mandell et al., 1998). Therefore, Awassi breed may produce more healthy and flavor meat than Najdi breed. The C18:1 constituted the highest percentage of MUFA in both breeds for intramuscular and subcutaneous fats. The value observed in this study was lower than the value obtained by Demirel et al. (2004) and Borys et al. (2012). These differences may be due to the difference in the breed (De Smet et al., 2004), type of feed offered to the animals (Bas et al., 2005) or carcass weight during slaughtering (Sanudo et al., 1998), Because, a change in diet after weaning and the increased slaughter weight influence significantly the fatty acid profiles (Dhanda et al., 2003; Beserra et al., 2004). The fatty acids represent the highest proportion of total fatty acids in intramuscular and subcutaneous fat are C16:0, C18:0 and C18:1 which is similar to the results obtained by Rowe et al. (1999) in lamb, Oka et al. (2002) in steers and Bonvillani et al. (2010) in goats. In term of C18:2, Awassi carcasses showed significantly higher percentage in Longissimus dorsi without significant difference in other muscles and subcutaneous fat. The value of C18:2 of both breeds obtained in this study are similar to those reported by Matsushita et al. (2010) in lamb and Oka et al. (2002) in steers. Potkanski et al. (2002) reported significant differences in the C18:2 in subcutaneous fats among breeds. Whereas, Cameron et al. (1994) found slight differences in the fatty acid content of the subcutaneous fat among breeds. Petrik et al. (2000b) suggested the importance of the dietary C18:2 on human health and Chin et al. (1992) noted that the C18:2 is not manufactured in human body using plant crops but can be obtained from the meat and milk. The intramuscular and subcutaneous fats among two breeds did not show significant differences in the percentage of CLA which disagreed with the results of Wachira et al. (2002) who found the CLA percentage in the Longissimus dorsi muscle and subcutaneous fat was significantly different among different lamb breeds. Najdi and Awassi breed did not show significant difference in term of C18:3 in intramuscular and subcutaneous fat. The result obtained in this study is similar to the finding of De Freitas et al. (2014) in the intramuscular fat of the muscle Longissimus dorsi in steers. Fisher et al. (2000) showed that semimembranosus muscle contained higher percentage of C18:3 compared to the value observed in this study and Caneque et al. (2001) showed higher percentage of C18:3 of intramuscular fat of Longissimus dorsi of lambs. This may be attributed to the use of different breeds (De Smet et al., 2004) or feeding on different diets (Bas et al., 2005; Lee et al., 2008). Also, an increase of C18:3 is considered beneficial to consumer health (Gill et al., 1995; Warnants et al., 1996). Although, the n-6/n-3 ratio in Awassi carcasses was significantly (p<0.05) higher in Longissimus dorsi but it was not significantly different between the two breeds in other muscles and subcutaneous fat. All the values obtained, except for the Longissimus dorsi of Najdi carcasses, were much higher than the values recommended for the human diet (≤4) (Enser et al., 1998). It is well recognized, biologically, that it is very important to maintain the relative balanced of n-6/n-3 in the human body. An increase of n-6/n-3 ratio may result in a high inflammation status in the body, which may be the cause of many diseases such as cardiovascular disease, autoimmune diseases and cancers (Simopoulos, 2008; Mozaffarian et al., 2005). Therefore, from nutritional standpoint, only Longissimus dorsi of Najdi breed is more healthy in term of n-6/n-3 ratio and other muscles and subcutaneous fat should be taken into major consideration when consumed by human. The PUFA/SFA ratio for Awassi and Najdi carcasses for intramuscular and subcutaneous fats was not significantly (p≥0.05) different. However, the ratio of PUFA/SFA for breeds was below the recommended value of 0.4 (Simopoulos, 2004). The result agrees with those of Enser et al. (1996) who showed that the PUFA/SFA ratio of subcutaneous adipose tissue and Longissimus muscle from lion chop or steaks of English sheep was 0.09 and 0.15, respectively. In contrast, this result is in disagreement with the findings of Oriani et al. (2005) who observed a significantly higher PUFA/SFA ratio in the semimembranosus muscle compared to Longissimus dorsi muscles in Italian lambs.
CONCLUSION
Since all the animals in this study were kept under same conditions, consumed similar diets, therefore the difference in the fatty acid composition would probably be of genetic origin. The results of the present study suggest that Awassi lambs produced healthier meat than Najdi lambs depending on the fatty acid composition in their intramuscular and subcutaneous fats. Also, the differences in the fatty composition were not enough to be of any significance at the market level. However, this information is useful especially for the Saudi society because Awassi breed may be used for the genetic improvement of fatty acid composition of carcass tissue lipids without increasing carcass fatness as well as for interpreting the reported differences in fatty acid composition between Awassi and Najdi breeds. Based on n6/n3 ratio, the result indicated that the feeding systems applied in many commercial farms in Saudi Arabia for lambs at 12 month of age need to be reviewed to produce meat of high nutritional value. In conclusion, the lamb producers should consider to supplement an inert fat source in feed to alter fatty acids composition of lamb meat.
ACKNOWLEDGMENT
I am pleased to extend my sincere thanks and appreciation to the Deanship of Scientific Research at King Faisal University for funding this research. Also, special thanks to Mr. Soleman Bonhaiah for his assisting during slaughtering time.