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Figure 1. Liver of P0.25 treatment |
Figure 2. Liver of P1.5 treatment |
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Useful effect of Phyllanthus amarus powder as a feed additive on chicken was evaluated at many using levels with respect to growth performance, liver histopathology and gut microflora. 250 seven day old Cobb 500 chicks (sex balanced, 172.2 ± 8.8 g initial body weight) were randomly assigned in a completely randomized design (CRD) to five diets containing: no Phyllanthus amarus powder in the feed (P0); 0.25% Phyllanthus amarus powder in the feed (P0.25); 0.5% Phyllanthus amarus powder (P0.5); 1% plant powder (P1) and 1.5% Phyllanthus amarus powder in the feed (P1.5).
Different levels of Phyllanthus amarus powder in the diets did not affect average daily weight gain, average weight gain, and feed conversion ratio of chickens (P>0.05). However, using this plant powder had tendency in affecting the weight after viscera removal, carcass weight and thigh/carcass ratio among chickens. P1 and P1.5 levels tent to reduce the weight after viscera removal and carcass weight compared to 0.25% and 0.5% level and no level groups (P=0.051 and P=0.092). Whilst, thigh/carcass ratio of P0.25 and P0.5 treatments tent to be lower than those of P0 and P1.5 groups (P=0.094). Average score of liver fatty degeneration was showed that no adding Phyllanthus amarus treatment had most severe fatty degeneration in chicken livers (1.67) while this plant powder could decrease this problem in using P. amarus groups (P=0.085).
In the modern world now, people are returning to nature due to the safety and the environmental friendliness and that is the reason why natural products are gaining their special values. In treatment, more and more research about medicinal plants are done for treating not only human diseases but also animal diseases. In Vietnam, the use of plants for curing and preventing some animal diseases as well as for healthy animals has been applied by farmers for long time. Among those plants, Phyllanthus sp. such as P. amarus, P.niruri, P. urinaria are famous for their treatment of liver, kidney diseases, acnes, ophthalmitis, snake bite, fever (Do Tat Loi 2004). Aqueous extract of Phyllanthus amarus had an antiviral activity against white spot syndrome virus (WSSV) in shrimp at the dose of 150 mg/kg of animal body weight (Balasubramanian et al 2007).
Another point is that P. amarus showed its natural antibiotic value with some bacteria causing diseases not only in human but also in animals. Ethanolic extract of this plant had an impressive 8 mm of growth inhibition zone against Salmonella typhi by checking agar cup diffusion method, just after the highest zone of ciprofloxacin (9 mm) (Oluwafemi and Debiri 2008). Another study also found that ethanolic extract of P. amarus could inhibit the growth of Staphylococcus aureus, Escherichia coli and Klebsiella Spp. (Adegoke et al 2010). Furthermore, some biochemical parameters could be changed by administrating orally P. amarus aqueous extract at doses of 50, 100 and 200 mg/kg body weight such as increasing glucose uptake, reducing aspartate amino transferases (AST) and alanine amino transfarase (ALT) (James et al 2009). Kodakandla Venkata Syamasundar et al (1985) found that phyllanthin and hypophyllanthin in P. niruri have the hepatoprotective function. This plant ethanol extract also showed the its hepatoprotective effect by reducing the content of thiobarbituric acid reactive substances (TBARS), glutathione level and the activities of antioxidant enzymes, glutathione peroxidase (GPx), glutathione-S-transferase (GST), superoxide dismutase (SOD) and catalase (CAT) and through liver histopathological analysis in mice with aflatoxin B1-induced liver damage (Naaz at al 2007). Moreover, this plant powder also showed the effective protectiveness against aflatoxicosis in broilers fed 100ppb aflatoxin B1 and kept the growth performance normally (Sundaresan et al 2007). From one trial of student has proved that 5g of its extract/kg feed reduced feed conversion ratio and significantly increased feed intake of pig as well as improved their resistance against pathologen (Pham Huu Phuoc and Nguyen Quang Thieu 2010, not yet published). While phyllanthin content in the extract and in P. amarus dry leave powder were not so much different (0.8 percent and 0.52 percent) (Phuong et al 2010; Tripathi et al 2006), extract price is still very high compared with powder and extraction is much more complicated than making powder. Therefore, the purpose of this experiment is to test the effect of adding some powder concentrations in the chicken diet in improving growth performance and protecting health.
The experiment was conducted to compare the effects of five levels of Phyllanthus amarus powder in the chicken diet in improving chicken growth performance by comparing:
Feed conversion ratio (FCR), average weight gain, average daily weight gain
Fasted live weight, weight after viscera removal, carcass weight, breast/carcass ratio, thigh/carcass ratio
Liver histopathology
Isolate total Samonella and total Clostridium in the gut
The experiment was taken place in experimental farm of Nong Lam University, Ho Chi Minh city, Vietnam. This area is in Southeastern Vietnam, with a tropical monsoon with rainy season from May to October and dry season from November to April. The average temperature is 28°C with high humidity. The duration of this study is 42 days, from August to October, 2011.
This experiment was designed as a completely randomized design (CRD) with 5 levels of Phyllanthus amarus powder and 5 replicates. Each replication will have about 9 to 10 chickens per cage (sex balanced). The treatments were:
P0: No Phyllanthus amarus powder in the feed
P0.25: 0.25% Phyllanthus amarus powder in the feed
P0.5: 0.5% Phyllanthus amarus powder in the feed
P1: 1% Phyllanthus amarus powder in the feed
P1.5: 1.5% Phyllanthus amarus powder in the feed
P. amarus powder was purchased and analyzed for phyllanthin, hypophyllanthin and niranthin at Instutute of Chemical Technology, Ho Chi Minh city, Vietnam. During the experiment, two batches of plant powder were used to mix with feed, the substance concentrations are in table 1.
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Table 1. Composition of Phyllanthus amarus extract |
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Sample |
DM (%) |
Hypophyllanthin (% of DM) |
Phyllanthin (% of DM) |
Niranthin (% of DM) |
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P. amarus Batch 1 |
92.5 |
0.352 |
0.458 |
0.433 |
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P. amarus Batch 2 |
93.7 |
1.41 |
1.0 |
1.29 |
Formulated feed for chickens will be calculated to meet their nutrient requirement (NRC 1994) and then mixed with P. amarus powder in order to form the expected diets according to the above treatments (Table 2 and Table 3). Completed feeds of two feeding periods and plant powder were analyzed for gross composition, including crude protein, ether extract, crude fibre, ash, Ca and P at the Department of Animal Nutrition, Nong Lam University (Table 4).
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Table 2. Ingredient composition of the basal diet 1 for chicken from 0 to 21st day, as fed |
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Ingredients |
Amount (%) |
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Corn |
50.210 |
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Soybean meal |
22.310 |
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Cassava chip |
10.000 |
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Meat and bone meal 50 |
6.117 |
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Fish meal 55 |
5.857 |
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Fish oil |
4.167 |
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DDGS corn |
0.440 |
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Premix for broiler |
0.250 |
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DL-methionin |
0.194 |
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Salt |
0.154 |
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Coccidosis medicine |
0.100 |
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Cholin Chloride 60 |
0.100 |
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L-Lysin-HCl |
0.091 |
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Phytase |
0.010 |
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L-threonin |
0.001 |
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Total |
100 |
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Table 3. Ingredient composition of the basal diet 2 for chicken from 21st to 42nd day, as fed |
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Ingredient |
Amount (%) |
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Corn |
57.893 |
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Soybean meal |
15.804 |
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Cassava chip |
10.000 |
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Meat and bone meal 50 |
7.430 |
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Fish oil |
4.500 |
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Fish meal 55 |
2.163 |
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Fish meal 60 |
1.137 |
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Premix for broiler |
0.250 |
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DL-methionin |
0.210 |
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Salt |
0.188 |
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L-Lysin-HCl |
0.159 |
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Coccidosis medicine |
0.100 |
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Cholin Chloride 60 |
0.100 |
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L-threonin |
0.056 |
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Phytase |
0.010 |
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Total |
100 |
Table 4. Chemical composition of the basal diet, analysed values (% of DM) |
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Dry matter |
Crude protein |
Ether extract |
Crude fibre |
Ash |
NaCl |
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Feed period 1 |
88.5 |
19.8 |
5.91 |
3.58 |
7.82 |
0.40 |
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Feed period 2 |
87.7 |
17.2 |
5.71 |
3.04 |
5.89 |
0.44 |
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P. amarus batch 1 |
90.7 |
13.2 |
- |
2.65 |
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P. amarus batch 2 |
91.11 |
19.4 |
- |
12.6 |
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250 seven day old Cobb 500 chicks (sex balanced, 172.2 ± 8.8 g initial body weight) were raised in the experiment. The chicks were randomly assigned to the wooden cages according to the experimental design. The experimental period was 35 days. Drinking water was offered ad libitum during this study. Hygienic condition during the feeding period was kept for the animals. They were be fed ad libitum daily at 05:30 am; 3 pm; 7pm; 12pm and 3am. Vaccination was undertaken based on farm vaccination schedule.
The animals were weighed three times at the beginning (seven day), the middle (21st day) and the end of feeding trial period (42nd day) to calculate average weight gain, average daily weight gain and feed conversion ratio (FCR). Six birds from each treatment (one per cage) were sacrificed to record fasted live weight, weight after viscera removal, carcass weight, breast/carcass ratio, thigh/carcass ratio. Gut samples were collected from treatments (six samples for one group) to isolate total Samonella and total Clostridium in animal guts. The liver were sectioned to observe lesions and biopsies carried out to examine the histopathology.
The data were analyzed by Excel and Analysis of variance (ANOVA) using the CRD procedure of the Minitab software (version 13.3). Sources of variation are: levels of Phyllanthus amarus powder and error.
Adding different levels of Phyllanthus amarus powder in the diets did not affect the growth performance of chicken. Average daily weight gain, average weight gain, and feed conversion ratio were not significantly different among treatments (P>0.05) (Table 5). However, Sundaresan et al (2007) found that 1% of adding Phyllanthus amarus powder in the diet of chicken fed aflatoxin B1 (100ppb) could significantly improve the FCR compared to the basal diet. Maybe, the occurrence of aflatoxin B1 in the diet made the plant powder show its effect clearly.
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Table 5. Effects of P.amarus levels on growth performance of chicken |
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P0 |
P0.25 |
P0.5 |
P1 |
P1.5 |
SEM |
Prob. |
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ADG (g/day) |
59.3 |
57.2 |
56.8 |
54.4 |
54.6 |
3.73 |
0.272 |
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Average weight gain (g) |
2050 |
1980 |
1966 |
1887 |
1891 |
126 |
0.268 |
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FCR |
1.9 |
1.91 |
1.96 |
1.98 |
1.99 |
0.095 |
0.512 |
Table 6 shows the different tendency in weight after viscera removal, carcass weight and thigh/carcass ratio among chickens given diets with different plant powder levels. High levels of powder in the feed (P1 and P1.5) tent to reduce the weight after viscera removal and carcass weight compared to lower level and no level groups (P=0.051 and P=0.092). Whilst, thigh/carcass ratio of lower plant powder level groups from 0.25 to 1 per cent in the feed tent to have lower ratios than P0 and P1.5 groups (P=0.094). Maybe, high levels of Phyllanthus amarus powder (1% and 1.5%) did not have good impact on some slaughter weight parameters of chicken.
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Table 6. Effects of P.amarus levels on chicken slaughter weight |
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P0 |
P0.25 |
P0.5 |
P1 |
P1.5 |
SEM |
Prob. |
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Fasted live weight (g) |
2343 |
2293 |
2291 |
2290 |
2206 |
101 |
0.276 |
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Weight after viscera removal (g) |
1990 |
1941.7 |
1948.3 |
1933.3 |
1823.3 |
91.5 |
0.051 |
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Carcass weight (g) |
1665 |
1648.3 |
1636.7 |
1623.3 |
1545 |
75.9 |
0.092 |
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Breast/carcass ratio |
0.342 |
0.354 |
0.355 |
0.360 |
0.348 |
0.016 |
0.446 |
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Thigh/carcass ratio |
0.335 |
0.313 |
0.327 |
0.324 |
0.338 |
0.015 |
0.094 |
Liver histopathology was recorded as liver fatty degeneration score (from 1 to 3 mark). Average score of liver fatty degeneration was presented in table 6. No adding Phyllanthus amarus treatment showed most severe fatty degeneration in chicken livers while this plant powder could decrease this problem in using P. amarus groups (P=0.085). This maybe due to the hepatoprotective effect via antioxidant defense enzymes such as superoxide dismitase (SOD) and catalase (CAT) and reduced glutathione (GSH) (Chatterjee and Sil 2007). Also, Phyllanthus amarus has liver regenerative effect against liver cell damage by increasing thymidine kinase activities (Chattopadhyay et al 2006).
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Table 7. Effects of P.amarus levels on liver histopathology record |
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P0 |
P0.25 |
P0.5 |
P1 |
P1.5 |
SEM |
Prob. |
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Fatty degeneration score |
1.67 |
0.67 |
1.17 |
0.67 |
0.83 |
0.671 |
0.085 |
Using 1.5% of Phyllanthus amarus powder in the diet made the chicken liver tender and swollen compared to the lower level group (0.25%).
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Figure 1. Liver of P0.25 treatment |
Figure 2. Liver of P1.5 treatment |
Table 8 shows that 1.5% Phyllanthus amrus powder in the feed seemed to have higher occurrence ration of total Salmonella and total Clostridium in the chicken gut samples (33.3 and 33.3%) compared to other treatments.
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Table 8. Effects of P.amarus levels on gut microflora of chickens |
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P0 |
P0.25 |
P0.5 |
P1 |
P1.5 |
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Occurrence ratio in the gut (%) |
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- Salmonella |
16.7 |
16.7 |
0 |
0 |
33.3 |
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- Clostridium |
0 |
0 |
16.7 |
16.7 |
33.3 |
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Highest amount |
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- Salmonella (102 cfu /g) |
100 |
2900 |
0 |
0 |
1600 |
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- Clostridium (10 cfu/g) |
0 |
0 |
40 |
90 |
50 |
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Lowest amount |
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- Salmonella (102 cfu /g) |
0 |
0 |
0 |
0 |
0 |
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- Clostridium (10 cfu/g) |
0 |
0 |
0 |
0 |
0 |
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Average amount |
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- Salmonella (102 cfu /g) |
16.7 |
483.3 |
0 |
0 |
316.7 |
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- Clostridium (10 cfu/g) |
0 |
0 |
6.7 |
15 |
10 |
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Using different levels of Phyllanthus amarus powder (0.25%, 0.5%, 1% and 1.5%) in the feed did not affect the growth performance of chicken. However, they helped to decrease the fatty degeneration in histopathology of liver, although 1.5% treatment made the liver tender and swollen.
High levels of Phyllanthus amarus powder (1% and 1.5%) in the diet tent to reduce the weight after viscera removal, carcass weight of chicken.
1.5% Phyllanthus amrus powder in the feed seemed to have higher occurrence ration of total Salmonella and total Clostridium in the chicken gut samples (33.3 and 33.3%) compared to other treatments.
The authors gratefully acknowledge the support for this research received from the MEKARN program financed by Sida.
