EFFECT OF ROOT OF ECLIPTA ALBA ON BIOCHEMICAL
PARAMETERS OF CLARIAS BATRACHUS
Mishra Preeti* and Gupta Seema
Department of Zoology, Government N.P.G. College of Science, Raipur 492010,
Chhattisgarh, India.
ABSTRACT
The aim of the current study was to assess the effect of
aqueous/ethanolic root extracts of Eclipta alba on biochemical
variables (total serum protein, albumin, globulin and A/G ratio) of
Clarias batrachus. Fishes of average weight (70-80 g) were randomly
divided in three groups (n=20) control and treated with 10 and 20 ppm
of root extracts (aqueous/ethanolic) up to 28 days. Serum was collected
on first, second, third and fourth week after treatment. ANOVA shows
a significant effect of treatment (p<0.01) on all variables except
albumin treated with ethanolic extract. Significant increase in treated
groups was observed in total serum protein, albumin and globulin at
almost all sampling points. Decreasing trend was noticed in A/G ratio
in both the extracts treated group at each sampling point. The results suggested that the root
extracts of E. alba may help in the activation of the immune system and may be beneficial in
aquaculture used as a natural immunostimulant.
KEYWORDS: Clarias batrachus, Eclipta alba root, extracts, biochemical parameters.
INTRODUCTION
Fishes are said to be a gold in the water and plays an important role in nation’s economy[1].
Aquaculture is one of the fastest growing industries to meet the food demand of huge
population[2].The main purpose of fish culture is to obtain the maximum yield of nutritive,
disease free and healthy fishes. When fishes are cultured artificially and the population
reaches highest level, the possibility of infections may increase that leads to significant loss
of the fish fauna. Therefore, the strengthening of the immune system by the use of harmless
substances is the best remedy for the production of healthy fishes.
Volume 5, Issue 7, 1935-1941. Research Article ISSN 2277– 7105
*Corresponding Author
Dr. Mishra Preeti
Department of Zoology,
Government N.P.G.
College of Science,
Raipur 492010,
Chhattisgarh, India. Article Received on 20 May 2016,
Revised on 10 June 2016, Accepted on 01 July 2016
Plants have been used since ancient time for treatment of many ailments and diseases.
Recently, plants have received more attention for their immune stimulating functions in fish
culture[3] and are used as an alternative of the drug and antibiotic to control fish diseases. The
medicinal plants have specific secondary metabolites by which they can stimulate the
immune system of any organism[4].
Biochemical parameters of blood help in detection of fish health[5]. Serum proteins are known
as circulating proteins, used in forming defensive molecules that help the body to fight
against infections. The serum proteins are spliced into two group viz. albumins and globulins.
Albumins are a predictor of health. Globulin has an important role as defensive
proteins.Various medicinal plants are believed to be useful in strengthening the immune
system of the organism.
Eclipta alba (L) family - Asteraceae, traditionally known as Bhringaraja, used as traditional
medicine in many countries. Its pharmacological activities viz., analgesic, antimicrobial,
antiviral, antifungal, antinociceptive, anti-inflammatory, antioxidant, anti-hyperglycemic,
hepatoprotective, immunomodulatory, hair growth and wound healing activities were
summarized by[6]. It carries several secondary metabolites like coumestans, alkaloids,
thiophanes, flavonoids, polyacetylene, triterpens and their glycosides.[7]
The above properties of E.alba disclosed its medicinal importance. Therefore, this study was
planned to investigate the effect of the two doses (10 and 20 ppm) of root extracts
(aqueous/ethanolic) of Eclipta alba on biochemical parameters of Clarias batrachus.Such
study may be quite worthy and useful in productuion of healthy fishes.
MATERIAL AND METHODS Preperation of plant extract
Eclipta alba was collected from nearby area of Raipur (CG) India. Roots were separated,
washed thoroughly with deionozed water to remove soil particles, shade dried at room
temperature and pulverized by electric blender. Soxhlet apperatus was used to extract the
plant root under study with water/e t h an ol a s a s o l v e nt[ 8 ]. The total yield was estimated
as 13.18 % (aqueous) and 11.00% (ethanol) from 15 g of root powder. The extracts were
stored at 4°C as stock solution. These extracts were used as per the requirement to
Experimental model
Fish - Experimental fish Clarias batrachus of average weight 70±5.0g were purchased from
local fish market. They were disinfected by 0.1% KMnO4[9] and acclimatized for one week
under laboratory conditions.
Experimental design
Fishes of mixed sexes were separated into three groups (n=20) of control received vehicle
only and treated with 10 & 20 ppm of root extract (aqueous/alcoholic) in circular cemented
tank containing 10 litter water. Test fishes were fed with goat liver ad libitum every alternate
day and water was changed before treatment.
Sample collection
Blood sample was collected from unanesthized experimental fish at every four successive
seven days in tubes without anticoagulant and allowed to clot at room temperature. The
serum was separated by spinning down in centrifuging at 3000 rpm for 15-20 minutes. The
supernatant was collected and stored in sterile vials at -20°C for further analysis.
Biochemical estimation
Total protein content was estimated using Bovine Serum Albumin[10] (BSA) as standard and
globulin was determined by biuret method. Albumin was calculated by substracting the
globulin value from total protein. Serum A/G ratio was estimated by the method of Reinhold
et al. (1950).[11]
Statistical analysis of data
The experimental data recorded were statistically analysed by two way ANOVA at p<0.05
and differences among means were inerpretated using DMRT.
RESULTS
Results of present study are presented in Table 1 (aqueous) and Table 2 (ethanolic) after
exposure of 10 and 20 ppm doses of root extract of E. alba on C. batrachus.
Progressive weekly increase in total protein, albumin and globulin and decreasing trends in
A/G ratio were observed. ANOVA detected a significant treatment effect (p< 0.01) on all
estimated variables in both the extracts treated groups except albumin exposed to ethanolic
extract. The duration and interaction between treatment and duration were found to be
DISCUSSION
The result of the present study revealed that the total serum protein and globulin level
increased significantly in both (aqueous/ethanolic) extracts treated group with whereas a little
bit increase in albumin level was noticed in the group treated with aqueous extract. Reducing
trend was observed in A/G ratio. The A /G ratio differed insignificantly in treated groups as
compared to control group.
The increased level of total serum proteins, albumins and globulins were also reported in
Labeo rohita fingerlings treated with Withaniasomnifera root powder[12] and enhanced level
of serum protein, albumin, globulin and declined A/G ratio fed with Achyranthus aspera
mixed diet[13]. In rats,[14] reported significantly increased total protein, globulin and dose
dependent increase in albumin level with aqueous root extract of Pelargonium reniforme and
increased production of total protein and no significant change in albumin level after
treatment with aqueous extract of Ocimum sanctum was reported by[15]. Similar findings were
reported in Catla catla treated with ethanolic extract of Cynodon dactylon mixed diets.[16]
Table: 1. Effect of root extract (aqueous) of E. alba on biochemical parameters of C.batrachus
Day
Treatment
ANOVA
Control 10ppm 20ppm
TSP (g/dl)
T** Dns T x Dns -0.11$ 07 5.12 ± 0.05 a C 5.42 ± 0.04 a B 5.53 ± 0.04 a A
14 5.14 ± 0.04 a B 5.48 ± 0.03 a A 5.56 ± 0.03 a A 21 5.13 ± 0.01 a B 5.55 ± 0.03 a A 5.61 ± 0.04 a A 28 5.17 ± 0.01 a B 5.53 ± 0.03 a A 5.58 ± 0.03 a A
Albumin(g/dl)
T** Dns T x Dns -0.05$ 07 2.09 ± 0.01 a B 2.14 ± 0.01 a A 2.15 ± 0.01 a A
14 2.08 ± 0.02 a C 2.15 ± 0.02 a B 2.20 ± 0.02 a A 21 2.08 ± 0.01 a B 2.16 ± 0.01 a A 2.19 ± 0.01 a A 28 2.10 ± 0.01 a B 2.16 ± 0.01 a A 2.14 ± 0.01 a AB
Globulin(g/dl)
T** Dns T x Dns -0.11$ 07 3.03 ± 0.05 a B 3.28 ± 0.03 a A 3.38 ± 0.04 a A
14 3.06 ± 0.05 a B 3.34 ± 0.03 a A 3.37 ± 0.03 a A 21 3.04 ± 0.01 a B 3.39 ± 0.02 a A 3.43 ± 0.04 a A 28 3.07 ± 0.01 a B 3.38 ± 0.03 a A 3.43 ± 0.03 a A
A/G ratio
T** Dns, T x Dns -0.03$ 07 0.69 ± 0.01 a A 0.65 ± 0.01 a B 0.64 ± 0.01 a B
14 0.68 ± 0.01 a A 0.65 ± 0.01 a B 0.65 ± 0.01 a B 21 0.68 ± 0.01 a A 0.64 ± 0.01 a B 0.64 ± 0.01 a B 28 0.68 ± 0.01 a A 0.64 ± 0.01 a B 0.63 ± 0.01 a B
Values are expressed as Mean ± SE. Means in a column followed by different lower case
letters and mean in a row followed by different capital letters are significantly different at 5%
Table: 2. Effect of E.alba root (ethanolic) extract on biochemical parameters of C. batrachus
Day Treatment
ANOVA
Control 10ppm 20ppm
TSP (g/dl)
07 5.18 ± 0.05 a B 5.50 ± 0.03 a A 5.61 ± 0.01 a A
T** Dns T x Dns -0.15$
14 5.13 ± 0.05 a B 5.58 ± 0.04 a A 5.66 ± 0.04 a A
21 5.22 ± 0.04 a B 5.63 ± 0.04 a A 5.68 ± 0.06 a A
28 5.20 ± 0.03 a B 5.64 ± 0.06 a A 5.65 ± 0.10 a A
Albumin(g/dl)
07 2.11 ± 0.05 a A 2.18 ± 0.07 a A 2.17 ± 0.01 a A
Tns Dns T x Dns -0.15$
14 2.00 ± 0.05 a B 2.18 ± 0.03 a A 2.18 ± 0.04 a A
21 2.14 ± 0.03 a A 2.17 ± 0.04 a A 2.15 ± 0.06 a A
28 2.07 ± 0.01 a A 2.17 ± 0.05 a A 2.13 ± 0.06 a A
Globulin(g/dl)
T** Dns T x Dns -0.12$
07 3.07 ± 0.07 a B 3.33 ± 0.04 b A 3.44 ± 0.04 a A
14 3.13 ± 0.06 a B 3.40 ± 0.04 ab A 3.48 ± 0.02 a A
21 3.09 ± 0.02 a B 3.46 ± 0.03 a A 3.53 ± 0.06 a A
28 3.14 ± 0.02 a B 3.47 ± 0.03 a A 3.52 ± 0.03 a A
A/G ratio
T** Dns T x Dns -0.06$
07 0.69 ± 0.03 a A 0.66 ± 0.03 a A 0.63 ± 0.00 a B
14 0.64 ± 0.02 a A 0.64 ± 0.01 a A 0.63 ± 0.01 a A
21 0.70 ± 0.01 a A 0.63 ± 0.01 a B 0.61 ± 0.02 a B
28 0.66 ± 0.01 a A 0.63 ± 0.01 a A 0.61 ± 0.01 a A
Values are expressed as Mean ± SE. Means in a column followed by different lower case
letters and mean in a row followed by different capital letters are significantly different at 5%
level by DMRT. *p<0.05; ** p<0.01;***p<0.001, ns= not significant, $ = LSD at 5% level
Enhanced level of serum protein was reported by[17] in Catla catla fed with Plumbago rosea
diet. Presence of flavonoids, glycosides and alkaloids in aqueous and ethanolic extract of root
was reported[18], which may be responsible for elevation in total protein and globulin.[19]
Enhanced levels of serum total protein, albumin, and globulin are related with a stronger
innate immune response in fishes[3]. Increased total protein and decreased A/G ratio is the
indicator of health. Increment in globulin level may indicate stimulation of immune response
by increasing defensive molecule[20]. The increased albumin level indicates that the extract of
E. alba may promote liver functioning. Decline in the A/G ratio of test fishes might be due to
CONCLUSION
The present study reveals enhanced biochemical parameters in C. batrachus when treated
with the two extracts (aqueous and ethanolic) of root of E. alba. The use of both the extract of
E.alba may benefit to boost the immune system and helps in production of healthy fishes.
ACKNOWLEDGEMENT
Authers are thankful to University Grant Commission for financial support.
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