Zweil, H., Kamel, K., AbdelRahman Zahran, S., AbdELRahman, M. (2015). Effects of Acacia Nilotica Leaf Extract Supplementation on Physiological Parameters and Antioxidant Activity in Growing Rabbits. Journal of the Advances in Agricultural Researches, 20(2), 216-227. doi: 10.21608/jalexu.2015.161389
Hassan Zweil; Kamel Kamel; Soliman AbdelRahman Zahran; Mohamed Hassan AbdELRahman. "Effects of Acacia Nilotica Leaf Extract Supplementation on Physiological Parameters and Antioxidant Activity in Growing Rabbits". Journal of the Advances in Agricultural Researches, 20, 2, 2015, 216-227. doi: 10.21608/jalexu.2015.161389
Zweil, H., Kamel, K., AbdelRahman Zahran, S., AbdELRahman, M. (2015). 'Effects of Acacia Nilotica Leaf Extract Supplementation on Physiological Parameters and Antioxidant Activity in Growing Rabbits', Journal of the Advances in Agricultural Researches, 20(2), pp. 216-227. doi: 10.21608/jalexu.2015.161389
Zweil, H., Kamel, K., AbdelRahman Zahran, S., AbdELRahman, M. Effects of Acacia Nilotica Leaf Extract Supplementation on Physiological Parameters and Antioxidant Activity in Growing Rabbits. Journal of the Advances in Agricultural Researches, 2015; 20(2): 216-227. doi: 10.21608/jalexu.2015.161389
Effects of Acacia Nilotica Leaf Extract Supplementation on Physiological Parameters and Antioxidant Activity in Growing Rabbits
1Department of Animal and Fish Production, Faculty of Agriculture (Saba Basha), University of Alexandria, Alexandria, Egypt.
2Agriculture Research Center, Animal Production Research Institute, Cairo, Egypt.
3Faculty of Agriculture Saba basha- Alexandria University
Abstract
In the present study the antioxidant activity of Acacia nilotica leaf in growing rabbits was performed. Thirty weaned V-line rabbits aged 35 days (5 weeks) weighed 510g±8.30 (mean±SE) used for the study which lasted for 84 days (12 weeks). Animals were divided equally and randomly into three groups (10 in each one). The first group was fed ad libitum a commercial pelleted diet, while the other groups (second and third) were fed the same diet plus administration with oral daily Acacia nilotica leaf extract at a dose 100 (low dose) and 200 mg /kg body weight (high dose) respectively for 7 weeks. Treatment with aqueous ethanolic leaf extract of Acacia nilotica resulted in significant (p < 0.05) increase in blood plasma total protein, albumin and globulin, while blood plasma aminotransferase (AST) and alanine- aminotransferase (ALT), alkaline phosphatase (ALP), urea, triglyceride (TG) and glucose levels were significantly decreased as compared to untreated group (control). Results showed that the Acacia nilotica leaves extract has significant antioxidant effect by increasing blood plasma Glutathione peroxidase (GPx) and catalase and decrease blood plasma thiobarbituric acid-reactive substances (TBARS) activity as compared to control. The effect of Acacia nilotica leaves extract showed better response as dose dependent. This potential activity of Acacia nilotica leaf might be due to the presence of its phytochemicals or the collective action of many active ingredients. It could be concluded that Acacia nilotica leaf extract treatment significantly improved physiological and antioxidant activity of growing rabbit and this improved was dose dependent.
Rabbit production is an important branch of animal production. Rabbit meat is of high quality and safety. The susceptibility of rabbits to various infection diseases and high mortality of young rabbits after weaning were studies. Frankič et al. (2009) reported that the main scope in animal husbandry to ensure good performance of farm animals and get quality animal products. In this aspect, herbs and spices are not just appetite and digestion stimulants, but can, with impact on other physiological functions, help to ensure good health and welfare of the animals, which could positively affect their performance. The use of phyto-additives and their extracts in rabbit husbandry offers an acceptable way to improve welfare and health (Szaboova, et al,. 2008).
Acacia contains variety of bioactive components such as phenolic acids, alkaloids, terpenes, tannins and flavonoids which are responsible for numerous biological and pharmacological properties like hypoglycemic, anti-inflammatory, anti-bacterial, anti-platelet aggregatory, anti-hypertensive, analgesic, anticancer, and anti-atherosclerotic due to their strong antioxidant and free radical scavenging activities (Sulaiman and Gopalakrishnan, 2011). Similar conclusion was reported by Seigler (2003) who reported that acacia species including amines and alkaloids, cyanogenic glycosides, cyclitols, fatty acids and seed oils, fluoroacetate, gums, non- protein amino acids, terpenes, hydrolysable tannins, flavonoids and condensed tannins.Phenolics are largest group of phytochemicals and accounts for most of the antioxidant activity in plants or plant products (Okpuzar, et al., 2009).
Sharma et al. (2014) investigated the antibacterial, antifungal, antiviral, and immunomodulatory potential of hot aqueous extract of Acacia nilotica leaves. They found that on dry matter basis, the filtered acacia nilotica leaves had a good extraction ratio (33.46%) and was found to have carbohydrates, glycosides, phytosterols, phenolic compounds, saponins, and flavonoids as major constituents. and these supports its use and availability in folk medicine. The efficacy of these compounds is already well established for antimicrobial activities (Mustafa, et al. 1999). Similar results were found of phytoconstituents of different extracts of Acacia nilotica, namely, aqueous extract (Kalaivani, et al. 2011), methanolic, and ethanolic extracts (Solomon-Wisdom and Shittu, 2010).
Thus, the present study was planned to study the possibly improvement effects of extract of Acacia nilotica leaves on physiological and antioxidant parameters in growing rabbits.
MATERIALS AND METHODS
The present study was carried out at the Rabbit Research Laboratory, in the Animal and Fish Production Department, Faculty of Agriculture (Saba Basha) during breeding season from October to February (winter, 2012).
Preparation of extract
Acacia nilotica Leaves (10g) were soaked and washed with plenty of water. The Acacia nilotica leaves extract was prepared in 100 ml of 80% aqueous methanol after crushing and macerating Acacia nilotica leaves (Alharbi and Azmat, 2011). After 3 days, the supernatant was completely removed by a boiling water bath at 45оC. The obtained residue was kept in the refrigerator for further use. The extract was made up to a known volume with distilled water just before oral administration.
Diets and animals
Thirty weaned V-line rabbits aged 35 days (5 weeks) and weighed 510g±8.30 (mean±SE) used for the study which lasted for 84 days (12 weeks) and were divided equally and randomly into three groups (10 in each one). The first group was fed ad libitum a commercial pelleted diet according to NRC (1977) recommendations and kept untreated and served as a control. The other groups (second and third) were fed the same diet plus administration with oral daily Acacia nilotica leaf extract at a dose 100 (Low dose) and 200 mg /kg body weight (High dose) respectively for 7 weeks. All the experimental animals were healthy and clinically free from internal and external parasites and were kept under the same management and hygienic conditions.
Experimental procedure
Blood samples were collected from the marginal ear vein every other week from five rabbits from each group. Plasma was separated by centrifugation at 4000 rpm for 20 minutes and kept -20oC until blood analysis.
Stored plasma samples were analyzed for total proteins, albumin, activity asparate-aminotransferase (AST) and alanine- aminotransferase (ALT), alkaline phosphatase (ALP), urea, triglyceride (TG), and glucose using commercial kits.
Blood plasma Thiobarbituric acid-reactive substances (TBARS) were measured in by using the method of Tappel and Zalkin (1959). Catalase (CAT) activity was determined using the Luck method involving the decomposition of hydrogen peroxide (Luck, 1974). Glutathione peroxidase (GPx) activity was assayed using the method of Chiu et al. (1976).
Data were analyzed as a completely randomized design (Steel and Torrie, 1981) using the general linear model procedure of SAS (1986). Means were statistically compared using least significant difference (LSD) test at 0.05 significance level (Steel and Torrie, 1981).
The following model was used:
Yijk = μ + ai + bj + abij + eijk
where Yijk, experimental observation; μ, overall mean; ai, treatment effect; bj, week effect; abij, interaction effect of treatment and week; eijk, random error.
Results and Dissection
Effects of acacia extract on physiological parameters:
Blood plasma total protein, albumin and globulin:
The data in Table (1) found that oral supplementation with low and high doses of Acacia nilotica extract cause significant increase in plasma total protein, albumin and globulin compared with unsupplemented group. The effect of weeks on pervious parameters was not significant.
The data in Table (1) showed that there was significant interaction between treatment and weeks in plasma TP, Alb and globulin (Glb). The highly significant values of plasma TP, Alb and Glb were obtained in the group supplemented with low dose of acacia extract at the eighth of week. Acacia nilotica leaves are very digestible and have high levels of protein (Fagg, 2001). Kannan et al. (2013) found that Acacia nilotica elevated plasma total protein in Wistar rats.
Wu and Tsai (2006) showed that mice fed chitosan (isolated from papaya latex), at the dose of 2.5 g/kg body weight increased serum immunoglobulin amounts. The increase of total protein in blood rabbits fed papaya latex may be associated with improvement of crude protein digestibility (El-Kholy et al., 2008).
Table (1): Overall means (mean ±SE) and the interaction of blood plasma total protein (TP), albumin (Alb) and globulin (Glb) of growing rabbits as affected by aqueous extract acacia nilotica leaves supplementation
Item
TP
Alb
Glb
(g/100ml)
(g/100ml)
(g/100ml)
Effect of Treatment (T)
Control
6.12
±
0.15b
4.01
±
0.09 b
2.11
±
0.15 b
Acacia LD (T1)
7.40
±
0.17a
4.46
±
0.10 a
2.94
±
0.13 a
Acacia HD (T2)
7.08
±
0.14a
4.48
±
0.06 a
2.60
±
0.15 a
P value
0.0001
0.0001
0.0004
Effect of Weeks (W)
Week1
6.61
±
0.10
4.15
±
0.07
2.45
±
0.11
Week3
6.92
±
0.17
4.33
±
0.04
2.58
±
0.17
Week5
6.95
±
0.22
4.43
±
0.12
2.52
±
0.23
Week7
6.99
±
0.35
4.34
±
0.16
2.65
±
0.23
P value
0.2685
0.1783
0.8408
Interaction (T*W)
T1*W1
6.30
±
0.17 e
3.93
±
0.16de
2.37
±
0.30cd
T1*W3
6.23
±
0.24e
4.26
±
0.02bcd
1.98
±
0.23cd
T1*W5
6.52
±
0.40ed
4.18
±
0.15cd
2.34
±
0.43cd
T1*W7
5.41
±
0.13f
3.66
±
0.20e
1.75
±
0.19d
T2*W1
6.66
±
0.16cde
4.22
±
0.04bcde
2.44
±
0.19cd
T2*W3
7.57
±
0.17ab
4.24
±
0.07bcd
3.33
±
0.10ab
T2*W5
7.20
±
0.17bcd
4.64
±
0.26ab
2.56
±
0.21c
T2*W7
8.19
±
0.40a
4.75
±
0.26a
3.44
±
0.20a
T3*W1
6.86
±
0.08bcde
4.31
±
0.08abcd
2.54
±
0.03c
T3*W3
6.94
±
0.05bcde
4.50
±
0.05abc
2.44
±
0.08cd
T3*W5
7.14
±
0.50bcd
4.47
±
0.21abc
2.67
±
0.55abc
T3*W7
7.37
±
0.26bc
4.62
±
0.05abc
2.75
±
0.31abc
P value
0.0014
0.0323
0.0460
a,b,c,d,e,f Means within a column not sharing similar superscripts are significantly different (P<0.05). W1,3,5,7 represent the weeks of treatment.
As Acacia nilotica leaves contains flavonoides, polyphenolic compounds, tannins, glycosides, organic acids and coumains (El-Shanawany, 1996), the anti-microbial activity of plant leaves might responsible for the anti-bactrial activity of plants. Increased globulin concentration may be an indication of increased immunity in the rabbits since the liver will be to synthesize enough globulins for immunologic action as mentioned by Summonu and Oloyede (2007).
Blood plasma ALT, AST and ALP
The data in Table (2) reported that supplemented growing rabbits with Acacia nilotica leave extract caused significant decreased in blood plasma AST, ALT and ALP compared with control group.
Table (2): Overall means (mean ±SE) and the interaction of blood plasma ALT, AST and ALP of growing rabbits as affected by aqueous extract acacia nilotica leaves supplementation
Item
ALT
AST
ALP
(mg/dl)
(mg/dl)
(mg/dl)
Effect of Treatment (T)
Control
29.3
±
0.40a
57.1
±
1.79 a
144.3
±
3.52 a
Acacia LD
26.5
±
0.43 b
50.1
±
1.77 b
116.4
±
4.58 b
Acacia HD
24.4
±
1.02 c
48.3
±
1.44 b
113.2
±
6.12 b
P value
0.0001
0.0001
0.0001
Effect of Weeks (W)
Week1
28.5
±
0.52 a
56.6
±
1.66 a
143.4
±
5.97 a
Week3
27.6
±
0.63 a
53.2
±
2.06 ab
125.5
±
5.36 b
Week5
25.7
±
1.10 b
51.1
±
2.02 bc
117.8
±
4.16 bc
Week7
25.0
±
1.11 b
46.5
±
2.10 c
111.8
±
7.85 c
P value
0.0004
0.0008
0.0001
Interaction (T*W)
T1*W1
28.4
±
0.82ab
56.9
±
3.66a
146.0
±
3.96a
T1*W3
29.4
±
0.75a
57.3
±
4.90a
150.3
±
2.50a
T1*W5
29.7
±
1.08a
58.3
±
3.61a
131.4
±
8.68ab
T1*W7
29.8
±
0.61a
56.0
±
3.18a
149.3
±
9.02a
T2*W1
28.5
±
0.42ab
57.5
±
2.66a
141.7
±
9.05a
T2*W3
27.3
±
0.29abc
51.6
±
3.59ab
115.9
±
5.98bc
T2*W5
25.2
±
0.58c
50.4
±
2.04abc
109.7
±
2.73c
T2*W7
24.9
±
0.86cd
41.0
±
0.55d
98.4
±
4.41cd
T3*W1
28.8
±
1.40ab
55.4
±
2.78a
142.5
±
6.55b
T3*W3
26.0
±
1.44bc
50.8
±
1.16ab
110.4
±
5.22bc
T3*W5
22.2
±
2.05ed
44.7
±
1.63bcd
112.2
±
5.32bc
T3*W7
20.4
±
0.85e
42.3
±
1.18cd
87.8
±
2.21d
P value
0.0013
0.0064
0.0097
a,b,c,d,e,f Means within a column not sharing similar superscripts are significantly different (P<0.05). W1,3,5,7 represent the weeks of treatment.
The results in Table (2) showed that the lowest significant values of the pervious parameters were obtained at the end of experimental period compared with the binging of treatment. The interaction between the treatment and time was shown in blood plasma ALT, AST and ALP where the lowest significant values was obtained in group three at the eighth week.
Aspartate transaminase (AST) and alanine transaminase (ALT) are enzymes associated with the conversion of amino acids to ketoacids. They are pathophysiological marker enzymes used to assess tissue damage (Sriram and Subramanian, 2011). Interestingly, Acacia nilotica did not show any such side effects. There was a decrease in plasma AST and ALT levels compared to control group in Acacia nilotica extract treated groups which indicated non-toxic and tissue protective nature of Acacia nilotica.
Blood plasma urea, TG and glucose
The data in Table (3) reported that supplemented growing rabbits with water Acacia nilotica leaves extract caused significant decreased in blood plasma TG and glucose levels compared with control group. No significant difference was found for blood plasma urea concentration. The results in Table (3) showed that the lowest significant values of blood plasma glucose and the highest blood plasma urea level was shown at the weeks7, compared with the binging of treatment. The significant interaction between the treatment and time was shown in blood plasma urea, TG and glucose, where the lowest significant values was obtained in group three at the eighth week for plasma TG and glucose.
Glucose is not only a necessary nutrient for the development and growth of food-producing animals, but also a potent signal molecular that regulates protein synthesis (Goichon et al., 2011). Therefore, the lower level of circulating glucose suggests a high efficiency of glucose and protein use for a healthy animal through nutritional perspectives (Yin and Cheng, 2003), which at least partially contributes to the improvement of growth performance and feed efficiency in poultry.
Effects of acacia extract on antioxidant parameters:
Data in Table (4) showed that administration with Acacia nilotica extract caused a significant decrease in blood plasma TBARS and significant increase in blood plasma GPx and CAT levels. The effect of Acacia nilotica extract was dose dependent. Acacia nilotica leaves extract for 8 weeks resulted in significant change in pervious parameters during experimental period.
The significant interaction between treatments and weeks was shown in blood plasma TBARS, GPx and CAT levels. The lowest significant interaction in TBARS and the highest significant interaction in GPx and CAT were obtained at weeks eight in high Acacia nilotica leaves extract dose treatment group. The results of the present study clearly indicated that aqueous extract of Acacia nilotica leaves exhibited higher antioxidant and free radical scavenging potentials.
Table (3): Overall means (mean ±SE) and the interaction of blood plasma urea, TG and glucose of growing rabbits as affected by aqueous extract of Acacia nilotica leaves supplementation
Item
Urea
TG
Glucose
(mg/dl)
(mg/dl)
(mg/dl)
Effect of Treatment (T)
Control
53.7
±
1.32
67.5
±
1.80a
133.0
±
2.9a
Acacia LD
53.1
±
1.52
59.2
±
1.53 b
109.9
±
2.7b
Acacia HD
53.2
±
0.62
48.5
±
2.39 c
108.3
±
3.4b
P value
0.8766
0.0001
0.0001
Effect of Weeks (W)
Week1
48.1
±
1.34c
61.9
±
1.84
123.9
±
3.6a
Week3
50.5
±
0.99c
59.9
±
2.68
118.3
±
3.7ab
Week5
53.7
±
1.18b
53.3
±
3.68
113.2
±
4.8b
Week7
61.1
±
1.01a
58.4
±
4.59
112.9
±
5.5b
P value
0.0001
0.0905
0.0340
Interaction (T*W)
T1*W1
46.3
±
1.22g
63.4
±
2.98abc
124.2
±
8.16abc
T1*W3
48.9
±
1.96gf
68.6
±
4.12ab
135.0
±
5.00ab
T1*W5
57.5
±
2.06bc
64.1
±
5.66abc
134.0
±
5.34ab
T1*W7
62.0
±
1.15ab
73.8
±
4.10a
138.6
±
3.46a
T2*W1
48.9
±
4.06fg
60.8
±
2.67bc
122.8
±
3.84bcd
T2*W3
48.1
±
0.14fg
58.2
±
3.64bc
110.5
±
2.05cde
T2*W5
50.9
±
1.79efg
55.2
±
4.51c
102.6
±
3.45e
T2*W7
64.7
±
0.72a
62.6
±
4.31abc
103.9
±
6.19e
T3*W1
49.0
±
1.08fg
61.4
±
4.32bc
124.7
±
7.25abc
T3*W3
54.5
±
0.40cde
53.0
±
3.73c
109.5
±
3.15de
T3*W5
52.6
±
1.16def
40.6
±
4.40d
103.0
±
6.26e
T3*W7
56.6
±
0.71cd
38.8
±
5.06d
96.1
±
2.53e
P value
0.0003
0.0131
0.0042
a,b,c,d,e,f,g Means within a column not sharing similar superscripts are significantly different (P<0.05). W1,3,5,7 represent the weeks of treatment.
Table (4): Overall means (mean±SE) and the interaction of blood plasma TBARS, GPx and CAT of growing rabbits as affected by aqueous extract Acacia nilotica leaves supplementation
Item
TBARS
GPx
CAT
(nmol/ml)
(U/ml)
(U/ml)
Effect of Treatment (T)
Control
1.643
±
0.04a
0.882
±
0.01c
1.275
±
0.01c
Acacia LD
1.218
±
0.02b
1.011
±
0.02b
1.395
±
0.02b
Acacia HD
1.177
±
0.02c
1.134
±
0.03a
1.507
±
0.03a
P value
0.0001
0.0001
0.0001
Effect of Weeks (W)
Week1
1.325
±
0.01b
0.938
±
0.01c
1.280
±
0.02c
Week3
1.416
±
0.06a
1.014
±
0.03b
1.416
±
0.04a
Week5
1.326
±
0.07b
1.065
±
0.05a
1.425
±
0.04a
Week7
1.317
±
0.10b
1.020
±
0.05b
1.448
±
0.04a
P value
0.0001
0.0001
0.0001
Interaction (T*W)
T1*W1
1.314
±
0.01c
0.934
±
0.03cd
1.286
±
0.02de
T1*W3
1.746
±
0.02b
0.924
±
0.01d
1.304
±
0.04de
T1*W5
1.674
±
0.07b
0.833
±
0.02e
1.253
±
0.02e
T1*W7
1.839
±
0.01a
0.838
±
0.03e
1.258
±
0.03e
T2*W1
1.337
±
0.03c
0.946
±
0.02cd
1.289
±
0.04de
T2*W3
1.299
±
0.01c
0.995
±
0.04c
1.384
±
0.03cd
T2*W5
1.204
±
0.02d
1.122
±
0.01b
1.453
±
0.07bc
T2*W7
1.032
±
0.02e
0.983
±
0.01cd
1.454
±
0.03bc
T3*W1
1.326
±
0.01c
0.935
±
0.02cd
1.265
±
0.05e
T3*W3
1.201
±
0.01d
1.122
±
0.03b
1.562
±
0.05ab
T3*W5
1.100
±
0.01e
1.240
±
0.02a
1.570
±
0.05a
T3*W7
1.079
±
0.03e
1.240
±
0.03a
1.632
±
0.02a
P value
0.0001
0.0001
0.0004
a,b,c,d,e Means within a column not sharing similar superscripts are significantly different (P<0.05). W1,3,5,7 represent the weeks of treatment.
Natarajan and Srinivasan (2015) reported that Acacia nilotica leaves for 6 weeks resulted in a marked decrease in plasma TBARS and marked increase in superoxide dismutase (SOD), CAT and GPx activities as compared to alloxan induced diabetic rats. They added that Acacia nilotica leaf treatment increased the antioxidants and may there by help to control free radicals, as Acacia nilotica leaf has been reported to be rich in flavonoids and phenolic compounds, well-known antioxidants and also to possess in vitro free radical scavenging and antioxidant activity (Kalaivani and Mathew, 2010).
In conclusion, Acacia nilotica leaf extract treatment significantly improved physiological parameters and antioxidant activity of growing rabbit and this improvement was dose dependent.
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