Livestock Research for Rural Development 29 (5) 2017 Guide for preparation of papers LRRD Newsletter

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Arechaatechu L. seed and Anredera cordifolia (Ten) Steenis leaf powder supplementation reduced serum transaminase in laying hens

Hanang Pawitan Marlani, Endang Kusumanti and Retno Murwani1

Faculty of Animal Science and Agriculture
1 Natural Product Laboratory - Laboratorium Terpadu, Diponegoro University Semarang 50275, Central Java Indonesia.


Arecha catechu L (bettle nut) and Anredera cordifolia (Ten) Steenis (“Binahong”) are Indonesia indigenous medicines with antimicrobial and wound healing activities respectively. However, they have not been known to be used for poultry. This research was conducted to evaluate the effect ofA. catechu seed and A. cordifolia leaves powder supplementation on the liver status of laying hens indicated by serum glutamate piruvate transaminase (GPT) and glutamate oxaloacetate transaminases (GOT). Forty eight of 42 weeks old laying hens were allocated randomly into 4 treatment groups i.e. no supplementation (T0), supplemented with 0.025% A. catechu seed and A. cordifolia leaf powder (T1), 0.05% A. catechu seed and A. cordifolia leaf powder (T2), 0.1% A. catechu seed and A. cordifolia leaf powder (T3). Each treatment consisted of 6 replicates with two hens per replicate. Supplementation was carried out by administering alternately A. catechu seed powder and A. cordifolia leaf powder for 18 days. Blood samples were collected after 6 and 18 days supplementation.

Serum GOT was not affected by supplementation either after 6 or 18 days. Serum GPT was not significantly different (P>0.05%) after 6 days supplementation but it was significantly reduced (P<0.01) after 18 days supplementation. As serum GPT is an indicator of liver cells damage, it can be concluded that supplementation of 0.1% A.catechu seed and A. cordifolia leaf powder up to 18 days might improve liver status of the laying hens.

Key words: beetle nut leaves, binahong seed, glutamate piruvate transaminase, glutamate oxaloacetate transaminase, liver status


Commercial laying hens start to produce eggs at about 20 weeks of age and egg production spans about 12 months after which production gradually declines. Liver is the centre organ of metabolism where egg yolk components are synthesized and then transported and deposited in the ovary (Zaheer 2015). As production time progresses it also exposes the hens to environmental and disease challenges which consequently increase liver work load. It is also common that commercial layers are infested with worms and coccidia during production time.  Such disease challenge can affect egg production and hens health (Kumar et al 2015). At the same time, increasing awareness of consumer on quality product has led the egg industry to reduce or omit in feed medication. Therefore there has been an increasing interest on the use of herbs and botanicals to maintain poultry productivity and health. In the following research we carried out preliminary study on the use of A. catechu seed and A. cordifolia leaf powder as feed additive in laying hens. Areca catechu , an indigenous medicinal plants, has been shown to have antimicrobial, antihelmintic, and antioxidant activity (Baby and Raphael 2014). The activities are potentially beneficial to reduce disease challenge. Anredera cordifolia leaves are known in our traditional practice to assist wound healing (Lim et al 2007, Prasetyo and Herihadi 2013) and therefore are beneficial for recovery after disease challenge or metabolic activity associated cell damage. On the other hand herbal supplementation or medication such as antibiotic could have negative effect on liver cells (Murwani and Bayuardhi, 2007; Murwani and Murtini 2009, Murwani et al 2011). Therefore we measured serum GPT and GOT as indicators of liver status of the supplemented laying hens. The liver uses these cytoplasmic enzymes to metabolize amino acids and to make proteins. When liver cells are injured, transaminases leak out into the bloodstream and thus increasing the levels.

Materials and methods

Collection of plant material and preparation of powder

Fresh dry A.catechu seed were collected from local market and the pericarp was discarded. Fresh A. cordifolia leaves were obtained from local farmers and air dried. Each of the dried plant materias were ground to a coarse powder and filtered through 25-50 mesh screen to obtain a homogenous powder. The dried plant powder was stored in refrigerated container for further use.

Evaluation of A. catechu seedand A. cordifolia leaf powder in laying hens

Forty two weeks old of 48 Isa Brown laying hens with an average body weight of 1.82 ± 0.17 Kg were obtained from small scale local layer farmers and allocated randomly into 4 treatment groups i.e. T0: control with no supplementation; T1: supplemented with 0.025%A. catechu seed and A.cordifolia leaf powder; T2: supplemented with 0.05%A.catechu seed and A. cordifolia leaf powder; T3: supplemented with 0.1%A.catechu seed and A. cordifolia leaf powder. Each group consisted of 6 replicates with two hens per replicate. Each hen was given 120 g commercial layer diet (17% protein, Charoen Pokpand) per day and free access to drinking water. Supplementation were carried out by alternate administration ofA. catechu seed powder for 3 days followed by A. cordifolia leaf powder for 3 days. The alternate supplementation was carried out for 18 days by mixing it in the diet. Blood samples were collected from brachial vein of one hen from each replicate after 6 and 18 days supplementation. Blood sampling was done in the morning prior to feeding and serum was separated and stored frozen until analyses of GPT and GOT.

Serum GPT and GOT measurement

The activity of serum glutamic pyruvic transaminase (GPT) and serum glutamic oxaloacetic transaminase (GOT) were measured by Kinetic method according to International Federation of Clinical Chemistry (Murwani and Bayuardhi 2007, Murwani 2009, Murwani et al 2011).

Experimental design and statistical analysis:

A Completely Randomized Design with 4 treatments and 6 replicates was employed. Each replicate consisted of 2 hens. Serum GPT and GOT were analyzed by ANOVA and when means were significantly different (p<0.01) Duncan’s multiple range test was carried out. To compare means of GPT and GOT after 6 and 18 days supplementation T-test was performed (p<0.01). Statistical analyses were done using Excel.

Results and discussion

The results showed that body weight of layers at the end of experiment were the same as the weight at the beginning. Serum GOT and GPT were presented in Table 1 and it showed that alternate supplementation ofA. catechu seed and A. cordifolia leaf powder for 6 or 18 days did not affect serum GOT of the hens. Serum GOT was originated not only from liver but also from heart, skeletal muscle, intestine, kidney, and brain tissues (Murwani et al 2011). Supplementation of the powder up to 18 days which did not affect serum GOT indicated that the active constituent of the powder up to 0.1% did not cause injury to the tissues. The same was true for serum GPT after 6 days supplementation. Interestingly however, after 18 days supplementation, serum GPT in control groups (no supplementation) was significantly higher than all supplemented groups. Serum GPT of all supplemented groups were significantly reduced compared to control group at the end of 18 days supplementation.

Table 1. Serum GOT and GPT of laying hens after 6 and 18 days alternate supplementation of A. catechu seed and A. cordifolia leaf powder







GOT (U/L) :

6 days







18 days













GPT(U/L) :

6 days







18 days

46.1 a

8.5 b

8.3 b

9.4 b


1.15 x 10-7






Means in the same row without common letter are different at P <0.01
, between 6 and 18 days : ns = not significant, sig= significant at P <0.01

Serum GPT was mainly originated from the liver and reduced enzyme activity could indicate the presence of liver cells regeneration. This regeneration could be due to the active compound of A.cordifolia leaves such as ursolic acid, oleanolic acid, saponin, alkaloid, flavonoid which stimulate cell proliferation and collagen synthesis (Bonte et al 2003, 2006, Lim et al 2007, Miladiyah and Prabowo 2012, Morikawa et al 2015). The leaves also contain protein which can improve blood flow and hence nutrients supply to tissue cells to speed replacement of the damage cells (Chuang et al 2007). However, this possibility must be further studied by histophatology. On the other hand an increase in serum GPT of hens with no supplementation could be due to an increase in hepatic work load of the 42 weeks old producing hens used in this study. The hepatic load was associated with liver metabolic activity in egg yolk synthesis and host defense against environmental challenge. Such hepatic load of host defense was shown by a study which found an elevated level of serum transaminases in hens infested with coccidia (El-Maksoud 2014).



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Received 17 November 2016; Accepted 15 March 2017; Published 1 May 2017

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