International Seminar sustainable utilization of coastal resources in tropical zone, 19-20 October,2016, Bengkulu, Indonesia

The Effect of Fermented Sauropus androgynus Leaf Extract on Fat Deposition, Toxicity, and Meat-Bone Ratio in Broiler Chickens Reared in Coastal Areas U. Santoso, Kususiyah and Y. Fenita Department of Animal Science, Faculty of Agriculture, Bengkulu University Jalan WR Supratman, Kandang Limun, Bengkulu, Indonesia

ABSTRACT

The present study was conducted to evaluate the effectiveness of fermented Sauropus androgynus leaf extract (FSALE) for replacing commercial feed additive in broiler chickens. Completely randomized design was used in the present study. Eighty broilers aged 14 days were distributed into 5 groups. Each treatment group consisted of 4 replicates, and each replicate consisted of 4 broiler chickens. The five treatments were as follows: 1) Control without fermented Sauropus androgynus leaf extract (FSALE); 2) Feed control plus 4.5 g FSALE/kg diet; 3) Feed control plus 9 g FSALE/kg diet; 4) Feed control plus 13.5 g FSALE/kg diet; 5) Feed control plus 18 g FSALE/kg diet. Experimental results showed that FSALE had no effect on abdominal fat, gizzard fat, fatty liver score (FLS), liver, spleen, gizzard, toxicity, meat, bone and skin weight. Although it was not significantly differnet, FSALE inclusion tended to reduce abdominal fat deposition. It could be cocluded that FSALE inclusion could replace commercial feed additive without reducing fat deposition, produced normal internal organ weights, and no toxicity was observed. Key words: Fermented Sauropus androgynus leaf extract, fat deposition, toxicity

INTRODUCTION There is a positive correlation between abdominal fat and carcass fat levels in broiler chickens, so it can be stated that to determine how much carcass fat contents can be measured by levels of abdominal fat. Prediction carcass fat content is very important, because consumers need low fat carcass. These consumer demands are rational, because there is evidence that consuming the carcasses with a high fat content will increase the concentration of fat in the blood. The high concentration of fat in the blood correlates positively with increased risk of atherosclerosis and coronary heart disease. Furthermore, a commercial feed additive containing antibiotics cause negative impacts on human health. Commercial feed additive, which contains antibiotics and synthetic compounds have a high side effect (Li and Vederas, 2009) such as damaging the hormonal and immune systems (Cao et al., 2004) and drug resistance (Fojo and Menefee, 2007). In addition, consumers also need a broiler carcass with a high proportion of meat with bone and skin proportion is relatively low. Furthermore, broilers reared in coastal areas exposed heat stress so that the production was declined. To overcome these problems, it should be added feed additive to the broiler diets, in particular of potential medicinal plants. One potential medicinal plants is katuk (Sauropus androgynus).

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International Seminar sustainable utilization of coastal resources in tropical zone, 19-20 October,2016, Bengkulu, Indonesia

Agustal et al. (1997) reported that Sauropus androgynus leaf contains 6 main compounds, namely monomethyl succinate and cis-2-methyl cyclopentanol acetate, benzoic acid, phenyl malonic acid, 2-pyrolidinon and methyl pyroglutamate. This medicinal herb is rich in flavonoids, phenolic compounds and antioxidants (Andarwulan et al., 2010; Rattanasena, 2012), β-carotene, iron (Santoso et al., 2015) and vitamin C (Santoso, 2014). Vitamin C is well established to have antistress and antioxidant properties. Sauropus androgynus leaf extract have antibacterial properties (Paul and Anto, 2011; Gothadam et al., 2010; Ariharan et al., 2013). The inclusion of Sauropus androgynus leaf extract increased the number of effective bacteria in gastrointestinal tract such as Lactobacillus sp. (Santoso, 2001c; Santoso et al., 2001) resulting in higher body weight (Santoso, 2001a). Samad et al. (2014) reported that the supplementation of Sauropus androgynus leaf to the diet diseases resistance and non-specific immune responses in grouper Epinephelus coioides. Santoso et al. (2010a,b, 2013) reported that the Sauropus androgynus leaf extract reduces the fat content of broiler meat. Therefore, the present study was conducted to evaluate the effect of fermented Sauropus androgynus leaf extract on fat deposition, toxicity and meat-bone ratio in broiler chickens reared in coastal areas. It was assumed that FSALE could replace the commercial feed additive, and reduce fat deposition in broiler chickens.

MATERIALS AND METHODS The extraction of fermented Sauropus androgynus leaf Sauropus androgynus leaf was fermented by Saccharomyces cerevisiae (Santoso et al., 2015a). The fermented leaf was then extracted with hot water at 90°C for 20 minutes as described by Santoso et al. (2005). Broiler treatments Completely randomized design was used in the present study. Eighty broiler chickens aged 15 days were distributed into 5 groups. Each treatment group consisted of 4 replicates, and each replicate consisted of 4 broiler chikens. The 5 treatments are as follows: 1) Broiler chickens were fed diet without supplementation of fermented Sauropus androgynus leaves extract (FSALE) as the control (P0) 2) Broiler chickens were fed diet supplemented 4.5 g FSALE/kg (P1) 3) Broiler chickens were fed diet supplemented 9 g FSALE/kg (P2) 4) Broiler chickens were fed diet supplemented 13.5 g FSALE/kg (P3) 306

International Seminar sustainable utilization of coastal resources in tropical zone, 19-20 October,2016, Bengkulu, Indonesia

5) Broiler chickens were fed diet supplemented 18.0 g FSALE/kg (P4) Experimental diets contain 19% crude protein and 3200 kcal ME/kg. (NRC, 1994). All broiler chickens were provided diet and drinking water ad libitum. Sampling At the end of the study (aged 35 days), 5 broiler chickens for each treatment group were slaughtered and abdominal fat, gizzard fat, liver, spleen, gizzard were then measured. The values of those variables were calculated by the following equation: the weight of variable/live weight x 100%. Fatty liver score was measured by comparing the color of the liver with the standar color from value of 1 to 5. Toxycity was calculated by the following equation: (liver + spleen) weight/live weight x 100%. Meat bone ratio was measured by comparing breast or leg meats with breast or leg bones. Data analysis All data were subjected to ANOVA (Toutenburg and Shalabh, 2009) and if significantly different was tested further by Duncan's multiple range test.

RESULTS AND DISCUSSION The effect of fermented Sauropus androgynus leaf extract on fat deposition is presented in Table 1. Experimental results showed that FSALE had no effect on abdominal fat, gizzard fat, fatty liver score (FLS). Although it did not significant, the inclusion of FSALE tended to reduce abdominal fat deposition. The higher inclusion would be lower abdominal fat deposition. The reduction of abdominal fat might reflect the reduction of carcass fat content. Santoso et al. (2001) reported that unfermented Sauropus androgynus leaf extract reduced abdominal fat deposition. Table 1. The effect of fermented Sauropus androgynus leaf extract on fat deposition Variable

P0

P1

P2

P3

P4

Abdominal fat, % BW

1.98

1.51

1.55

1.59

1.35ns

Gizzard fat, % BW

0.51

0.62

0.71

0.53

0.57ns

Fatty liver score

2.81

2.12

2.75

2.37

2.44ns

P0= Broiler chickens were fed diet without supplementation of fermented Sauropus androgynus leaves extract (FSALE) as the control; P1 = Broiler chickens were fed diet supplemented 4.5 g FSALE/kg; P2 = Broiler chickens

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International Seminar sustainable utilization of coastal resources in tropical zone, 19-20 October,2016, Bengkulu, Indonesia

were fed diet supplemented 9 g FSALE/kg; P3 = Broiler chickens were fed diet supplemented 13.5 g FSALE/kg; P4 = Broiler chickens were fed diet supplemented 18.0 g FSALE/kg.

The effect of fermented Sauropus androgynus leaf extract on liver, spleen, gizzard and toxicity are presented in Table 2. Experimental results showed that FSALE had no effect on liver, spleen, gizzard and toxicity (P>0.05). Table 2. The effect of fermented Sauropus androgynus leaf extract on liver, gizzard and spleen weights, and toxicity Variable P0 P1 P2 P3 P4 Liver, % BW

2.13

1.92

2.29

1.82

2.01ns

Spleen, % BW

0.11

0.12

0.11

0.14

0.15ns

Gizzard, % BW

1.58

1.56

1.73

1.58

1.59ns

Toxicity

2.25

2.05

2.40

1.96

2.16ns

P0= Broiler chickens were fed diet without supplementation of fermented Sauropus androgynus leaves extract (FSALE) as the control; P1 = Broiler chickens were fed diet supplemented 4.5 g FSALE/kg; P2 = Broiler chickens were fed diet supplemented 9 g FSALE/kg; P3 = Broiler chickens were fed diet supplemented 13.5 g FSALE/kg; P4 = Broiler chickens were fed diet supplemented 18.0 g FSALE/kg.

Table 3. The effect of fermented Sauropus androgynus leaf extract on meat, bone and skin weight Variable P0 P1 P2 P3 P4 Breast meat, % BW

18.90

17.92

17.66

17.06

18.36ns

Breast bone, % BW

2.81

2.89

2.91

3.16

2.95ns

Breast skin, % BW

2.97

2.43

2.47

2.51

2.69ns

Meat

7.05

6.70

6.56

5.51

6.33ns

Leg meat, % BW

16.08

16.12

15.64

16.53

15.04ns

Leg bone, % BW

3.68

4.34

3.72

3.68

3.88ns

Leg skin, % BW

2.29

2.66

2.47

2.62

2.24ns

Meat bone ratio of leg

4.40

3.90

4.22

4.5

3.92ns

Breast, % BW

25.34

23.42

23.54

23.82

25.07ns

Leg, % BW

22.79

23.45

22.90

23.71

22.64ns

bone ratio of

breast

P0= Broiler chickens were fed diet without supplementation of fermented Sauropus androgynus leaves extract (FSALE) as the control; P1 = Broiler chickens were fed diet supplemented 4.5 g FSALE/kg; P2 = Broiler chickens were fed diet supplemented 9 g FSALE/kg; P3 = Broiler chickens were fed diet supplemented 13.5 g FSALE/kg; P4 = Broiler chickens were fed diet supplemented 18.0 g FSALE/kg.

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International Seminar sustainable utilization of coastal resources in tropical zone, 19-20 October,2016, Bengkulu, Indonesia

The present study showed that the inclusion of

FSALE produced normal liver,

gizzard and spleen weights. Santoso et al. (2015b) also reported that feeding fermented Sauropus androgynus powder had no effect on liver, gizzard and spleen weights. From the data of toxicity, it appears that FSALE is not toxic in broiler chickens. This study is in agreement with the observation of Santoso (2001c) and Santoso et al. (2015b). The effect of fermented Sauropus androgynus leaf extract on meat, bone and skin weight are presented in Table 3. Experimental results showed that FSALE had no effect on meat, bone and skin weight. Breast meat ranged from 17.06% to 18.90%, whereas leg meat ranged from 15.04% to 16.53%. Meat bone ratio of breast was higher that that of leg. This means that breast has more meat. The study showed that FSALE resulted in similar meat production as compared to the control group. Santoso (2001b) and Santoso et al. (2015b) reported that unfermented Sauropus androgynus leaf extract did not affect meat bone ratio of breast and leg in broiler chickens.

CONCLUSION It was concluded that supplementation of fermented Sauropus androgynus leaf extract could be used to replace commercial feed addtive without reducing the ratio of meat to bone. This extract did not cause toxicity to the broiler chickens as indicated by the same toxicity score at all treatment groups. The inclusion of the extract at 18 g/kg diet tended to reduce abdominal fat deposition.

REFERENCES Agustal, A., M. Harapini, and Chairul. 1997. Analisis kandungan kimia ekstrak daun katuk (Sauropus androgynus (L) Merr dengan GCMS. Warta Tumbuhan Obat Indonesia 3 (3): 31-33. Andarwulan, N., R. Batari, D. A. Sandrasari, B. Bolling, and H. Wijaya. 2010. Flavonoid content and antioxidant activity of vegetables from Indonesia. Food. Chem., 121:1231-1235. http://dx.doi.org/10.1016/j.foodchem.2010.01.033. Ariharan, V. N., V. N. M. Devi, and P. N. Prasad. 2013. Antibacterial activity of Sauropus and rogynous leaf extracts againts some pathogenic bacteria. Rasayan J. Chem. 6:134-137. Cao, J., K. Li, X. Lu, and Y. Zhao. 2004. Effects of florfenical and chromium (III) on humoral immune response in chicks. Asian-Aust. J. Anim. Sci. 17: 366-370. http://dx.doi.org/10.5713/ajas.2004.366 Fojo, T., and M. Menefee. 2007. Mechanisms of multidrug resistance: the potential role of microtubule-stabilizing agents. Annals of Oncology 18 (Supplement 5): v3-v8. Gothadam, K. M., R. Aishwarya, and S. Karthikeyan. 2010. Preliminary screening of antimicrobial properties of few medicinal plants. Journal of Phytology 2:1-6. 309

International Seminar sustainable utilization of coastal resources in tropical zone, 19-20 October,2016, Bengkulu, Indonesia

Li, J. W. H., and J. C. Vederas. 2009. Drug discovery and natural products: End of an era or an endless fontier? Science 325:161-165. National Research Council (NRC). 1994. Nutrient Requirement of Poultry. Ninth Revised Edition. National Academiy Press. Washington, D. C. Paul, M. and K. B. Anto. 2011. Antibacterial activity of Sauropus androgynus (L.) Merr. Int. J. Plant Sci., 6:189-192. Rattanasena, P. 2012. Antioxidant and antibacterial acitivities of vegetables and fruits commonly consumed in Thailand. Pak. J. Biol. Sci., doi:10.3923/pjbs.2012 Samad, A. P. A,, U. Santoso, M. C. Lee, and F. H. Nan. 2014. Effects of dietary katuk (Sauropus androgynus L. Merr.) on growth, non-specific immune and diseases resistance against Vibrio alginolyticus infection in grouper Epinephelus coioides. Fish Shellfish Immunol. 30:582-589. http://dx.doi.org/10.1016/j.fsi.2013.11.011 Santoso, U. 2001a. Effect of Sauropus androgynus Extract on the carcass quality of broiler chicks. Buletin Ilmu Peternakan dan Perikanan 7: 22-28. Santoso, U. 2001b. Effect of Sauropus androgynus extract on the performance of broiler. Buletin Ilmu Peternakan dan Perikanan 7: 15-21. Santoso, U. 2001c. Effect of Sauropus androgynus extract on organ weight, toxicity and number of Salmonella sp and Escherichia coli of broilers meat. Buletin Ilmu Peternakan dan Perikanan, 7 (2): 162-169. Santoso, U., Suharyanto, and E. Handayani. 2001. Effects of Sauropus androgynus (katuk) leaf extract on growth, fat accumulation and fecal microorganisms in broiler chickens. Jurnal Ilmu Ternak dan Veteriner 6:220-226. Santoso, U., J. Setianto and T. Suteky, 2005. Effects of Sauropus androgynus (katuk) extract on egg production and lipid metabolism in layers. Asian-Australasian J. Anim. Sci., 18: 364-369. http://dx.doi.org/10.5713/ajas.2005.364 Santoso, U., Kususiyah, and Y. Fenita. 2010a. The effect of Sauropus androgynus extract and lemuru oil on fat deposition and fatty acid composition of meat in broiler chickens. J. Indonesian Trop. Anim. Agric., 35:48-54. http://dx.doi.org/10.14710/jitaa.35.1.48-54 Santoso, U., T. Suteky, and Y. Fenita. 2010b. Effects of supplementation of alkaloid and non alkaloid from Sauropus androgynus leaves on egg production and lipid profil in layer chicken. J. Anim. Prod. 12:184-189. http://dx.doi.org/10.20884/1.anprod.2010.12.3.290 Santoso, U., Kususiyah, and Y. Fenita. 2013. Effect of Sauropus androgynus leaves extract in fat deposition in broilers fed low containing diets. J. Indonesian Trop. Anim. Agric., 38:176-184. http://dx.doi.org/10.14710/jitaa.38.3.176-184 Santoso, U. 2014. Katuk, Tumbuhan Multi Khasiat. Badan Penerbit Fakultas Pertanian, Unib, Bengkulu. Santoso, U., Y. Fenita, Kususiyah, and I. G. N. G. Bidura. 2015 a. Effect of fermented Sauropus androgynus leaves on meat composition, amino acid and fatty acid compositions in broiler chickens. Pak. J. Nutr. 14:799-807. http://dx.doi.org/10.3923/pjn.2015.799.807 Santoso, U., Y. Fenita and Kususiyah, 2015b. The Effect of Fermented Sauropus androgynus Leaves on Performance, Fat Deposition and Carcass Quality in Broiler Chicken. International Seminar on Promoting Local Resources for Food and Health, October 12-13, 2015, Bengkulu University, Bengkulu, Indonesia Toutenburg, H. and H.T. Shalabh. 2009. Statistical Analysis of Designed Experiments. 3 rd ed. Springer Science+Business Media, LLC. New York, Dordrecht, Heidelberg, London.

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