Water soluble fraction of Tinospora cordifolia leaves enhanced the non-specific immune mechanisms and disease resistance in Oreochromis mossambicus
Introduction
The intensive culturing of fish generates a stressful environment to them and the consequent suppression of the immune system, rendering the fish susceptible to the infectious diseases. To prevent financial losses due to diseases, fish farmers must take sufficient preventive measures. Antibiotics and other chemotherapeutics used to control these diseases can result in the development of drug resistant pathogens, environmental pollution and accumulation of residues in fish. Though vaccination is an effective prophylactic method for controlling fish diseases, vaccines are relatively expensive and since they are very specific to particular pathogens [1], there is a need for developing and using number of vaccines against corresponding number of microbial diseases against which the fish have to be protected. But the use of immunostimulants for the disease prevention in fish is considered as a promising alternative to vaccines [2], [3] because of its broad spectrum activity and so its cost effectiveness.
Recently, growing interest has been paid to the immune stimulating function of medicinal herbs in aquaculture. The herbal immunostimulants which have been tested includes saponin [4], glycyrrhizin [5], aloe [6], Ocimum sanctum extracts [7], [8], azadirachtin [9], [10], Oro glo layer dry, a xanthophyll preparation from Marigolds [11], Viscum album, Urtica dioica and Zingiber officinale [12], Radix astragalin seu Hedysari and Radix angelicae sinensis [13], [14], Astragalus radix and Scutellari radix [15] and Achyranthes aspera [16], [17], [18], Solanum trilobatum [19] and Eclipta alba [20] extracts.
Tinospora cordifolia is a large, glabrous, deciduous climbing shrub belonging to the family Menispermaceae. It is distributed throughout tropical Indian subcontinent and China, upto an altitude of 300 m [21]. In Indian Ayurvedic system of medicine, the powder and aqueous extract of the plant is used in debility, hepatitis, dyspepsia, jaundice and other liver afflictions. Its antidiabetic [22], anti-inflammatory [23] and diuretic activities [24] have also been reported. A variety of constituents have been isolated from T. cordifolia belonging to different classes such as alkaloids, diterpenoid lactones, glycosides, steroids and sesquiterpenoids [25]. Some of the active principles of T. cordifolia were found to possess immunomodulatory activity in mice [26]. Although a few studies on the immunomodulatory effect of T. cordifolia in mammalian species have been reported [27], [28], this is probably the first detailed study in fish on the effect of water soluble fraction of T. cordifolia leaves on the non-specific immune mechanisms and disease resistance in Mozambiquetilapia, Oreochromis mossambicus against Aeromonas hydrophila.
Section snippets
Experimental animal and their maintenance
O. mossambicus, an exotic freshwater teleost fish was used in this study. Male fish weighing 25 ± 5 g (for non-specific humoral assays and disease resistance studies) and 50 ± 5 g (for non-specific cellular assays) were obtained from a local fish farmer and acclimated for two weeks in 150 L fiber reinforced plastic (FRP) tanks. They were kept at the ambient, uncontrolled temperature of 28 ± 2 °C under natural photoperiod. Canister external water filters (Eheim, Germany) were used to maintain
Serum lysozyme activity
As shown in Fig. 1, the serum lysozyme activity measured in units ml−1 was higher in all the treated groups than the untreated control group on all the days tested. The difference was highly significant (P < 0.001) throughout the study period. The maximal enhancement of lysozyme activity was observed on day 6 in the 6 and 60 mg kg−1 treated groups (833.58 units ml−1, P < 0.001; 737.3358 units ml−1, P < 0.001) and on day 8 in the 600 mg kg−1 treated group (656.5 units ml−1, P < 0.001). Among the
Discussion
The present study revealed that intraperitoneal administration of water soluble fraction of T. cordifolia leaf significantly increased the non-specific humoral and cellular responses and disease resistance against A. hydrophila.
Lysozyme level is a measurable humoral component of the non-specific defence mechanisms. Lysozyme hydrolyzes β-1-4-glucosidic linkages between N-acetylmuramicacid and N-acetyl-d-glucosamine residues present in the mucopolysaccharide cell wall of a variety of bacterial
Acknowledgements
The financial support of the University Grants Commission, New Delhi for the investigation is gratefully acknowledged.
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