• No results found

EFFECT OF ROOT OF ECLIPTA ALBA ON BIOCHEMICAL PARAMETERS OF CLARIAS BATRACHUS

N/A
N/A
Protected

Academic year: 2020

Share "EFFECT OF ROOT OF ECLIPTA ALBA ON BIOCHEMICAL PARAMETERS OF CLARIAS BATRACHUS"

Copied!
7
0
0

Loading.... (view fulltext now)

Full text

(1)

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

(2)

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

(3)

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

(4)

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%

(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

(6)

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.

REFERENCES

1. Jawale S, Fartade A, Borde S. Biochemical studies on Caryophyllidean tape worms in

fresh water fish Clarias batrachus. Recent Research in Science and Technology, 2011; 3:

35-36.

2. FAO. The state of World Fisheries and Aquaculture (SOFIA), 2004.

3. Wiegertjes G F, Stet R J M, Parmentier H K, Van Muiswinkel W B, “Immunogenetics of

disease resistance in fish: A comparative approach”. Develop. Compar. Immuno., 1996;

20: 365-381.

4. Ardo L, Yin G, Xu P, Varadi L, Szigeti, G, Jeney Z, Jeney G. Chinese herbs (Astragalus

membranaceus and Lonicera japonica) and boron enhance the non specific immune

response of Nile Tilapia (Oreochromis niloticus) and resistance against Aeromonas

hydrophila. Aquaculture, 2008; 275: 26-33.

5. De Pedro N, Guijarro A E, Lopez-Patino M A, Martinez- Alvarez R, Delgado M. Daily

and seasonal variation in haematological and blood biochemical parameters in tench

Tinca tinca. Aquaculture, Res., 2005; 36: 85-96.

6. Craig W J. Health promoting properties of common herbs. Am. Journal of Clinical

Nutrition, 1999; 70: 491S-9S.

7. Neeraja PV, Margaret E. Eclipta alba (L.) Hassk: A valuable Medicinal herb. IJPCR,

2012; 2:4: 188-197

8. Gupta S., Mishra P., Effect of leaf extract of Eclipta alba on hematological parameters of

Claras gariepinus (Burchell, 1822).World Journal of Pharmaceutical Research, 2014;

3(9): 4860-4870.

9. Joshi P K, Harish D, Bose M. Effect of lindane and malathione exposure to certain blood

parameters in a fresh water teleost fish Clarias batrachus. Pollution Resources, 2002; 21:

(7)

10.Lowry O H., Rosebrough N J., Farr A L., Randall R J., Protein measurment with the folin

phenol reagent. J.Biol. Chem., 1951; 193: 265-275.

11.Oser B L. Hawk’s physiological chemistry. 14 ed., Tata McGraw Hill Publishing Co.

Ltd., New Delhi, 1965.

12.12. Sharma A, Deo A D, Tandel Riteshkumar S, Chanu T I, Das A. Effect of Withania

somnifera (L. Dunal) root as a feed additive to immunological parameters and disease

resistance to Aeromonas hydrophila in Labeo rohita (Hamilton) fingerlings. Fish and

Shellfish Immunology, 2010; 29: 508-512.

13.Rao V Y, Das B K, Jyotymayee P, Chakrabarti R. Effect of Achyranthes aspera on the

immunity and survival of Labeo rohita infected with Aeromonas hydrophila. Fish &

Shellfish Immunology, 2006; 20: 263-273.

14.Adewusi E A, Afolayan A J. Safety evaluation of the extract from the roots of

Pelargonium reniforme Curtis in male wistar rats. African Journal of Pharmacy and

Pharmacology, 2009; 3: 368-373.

15.Jeba R C, Vaidyanathan R, Rameshkumar G. Immunomodulatory activity of aqueous

extract of Ocimum sanctum in rat. International Journal of Pharmaceutical and

Biomedical Research(IJPBR), 2011; 2: 33-38.

16.Kaleeswaran B, Ilavenil S, Ravikumar S. Changes in biochemical, histological and

specific immune parameters in Catla catla (Ham.) by Cynodon dactylon (L.) Journal of

King Saud University (Science), 2012; 24: 139-152.

17.Innocent B X, Fatima M S A, Siva rajani S. Immune response of Catla catla fed with oral

immunostimulant Plumbago rosea and post challenged with Aeromonas hydrophila.

International Journal of Applied Pharmaceutical Technology, 2011; 2: 447-454.

18.Mishra P, Gupta S. Phytochemical Screening of Eclipta alba. Asian Journal of Animal

Science, 2013; 8(2): 112-114.

19.Rajendran R, Hemlatha S, Akasakalai K, Madhukrishna C H, Sohil B, Sundaram R M.

Hepatoprotective activity of Mimosa pudica leaves against Carbon tetra chloride induced

toxicity. Journal of Natural products, 2009; 2:116-122.

20.Puri A, Saxena R, Saxena R P, Saxena K C, Srivastava V, Tandon J S. Immunostimulant

References

Related documents

present study, the inhibition of the mTOR signaling pathway, SIVA, and cell proliferation, apoptosis was observed when treated with plumbagin in HCC cells. To investigate whether

Section II contained questions (mostly multiple-choice) on the approaches used to diag- nose and treat infections (for example, use and character- istics of surveillance

of 1 also showed the presence of an anomeric proton as a doublet at δ 5.40 suggesting a sugar residue in its structure which was identified as L-rhamnosyl

This approach utilizes a clinic visit spread out over 1 day or 2 days in which the patient is scheduled for radiographic studies, laboratory tests, history and physical

The first chapter briefly introduces the background, objective and contribution, making clear the structure of the research. And the second chapter elaborates the major theories

Conclusions: Excessive daytime sleepiness in First Nations peoples living on reserves in rural Saskatchewan is associated with factors related to respiratory co-morbidities,

Figure 1 Fractional composition of phospholipids from bronchoalveolar lavage fluid BALF), tracheal wash (TW) and induced sputum (IS).. PC, phosphatidylcholine; PG,

aspirin in patients at risk of ischemic events; CARDIA: coronary artery risk development in young adults; CARDS: Collaborative Atorvastatin Diabetes Study; CCBs: calcium