Research Journal of Animal, Veterinary and Fishery Sciences ___________________________ ISSN 2320 – 6535 Vol. 2(5), 1-5, May (2014) Res. J. Animal, Veterinary and Fishery Sci. International Science Congress Association 1 A Study on the Immunoprotective effect of the Medicinal Plant Aloe Vera on the Common Carp Cyprinus Carpio (L) Valsa Judit Anto A. and V. Balasubramanian Post Graduate and Research Department of Zoology, Ayya Nadar Janaki Ammal (Autonomous) College, Sivakasi, INDIA Available online at: www.isca.in, www.isca.me Received 8th April 2014, revised 10th May 2014, accepted 22nd May 2014 AbstractThe present study has been carried out to evaluate the immuno stimulatory effect of Aloe vera. A.vera is an active compound and it contains protein, amino acids, and enzymes. Different concentrations (250,500,750mg/kg of food) of extract of Aloe vera were fed to common carp, Cyprinus carpio for 30 days. Fish pathogen, Aeromonas hydrophila with 0.1ml of 10CFU/ml was injected intraperitonialy. The cumulative percentage mortality, oxygen consumption and opercular movement were calculated and observed. After the treatment, cumulative percentage mortality, increased opercular movement and oxygen consumption were observed. Thus Aloe vera acts as an immunostimulant to enhance the activity of fish. Keywords: Cyprinus carpio, Aeromonas hydrophila, Aloe vera, cumulative percentage mortality, Opercular movement, Oxygen consumption. IntroductionImmunostimulants are valuable for the prevention and control of fish disease in aqua culture as they represent an alternative and supplementary treatment to vaccination. The immunostimulants also have additional effects such as growth enhancement and increase in the survival rates of the fishes under stress. The common carp C.carpio is commercially an important fish species in aquaculture. India is rich in rivers, ponds and streams whose major fish fauna consists of C.carpio. The aquaculture products were serving as a stable food for people. Riverine fishery is one of the essential economic components of the fishery sector of any state. The fish can eat vegetarian diet of water plants but prefer to scavenge the bottom for insects, crustaceans, crawfish, and benthic worms. The common practice of fish importation for aquaculture was in the form of fingerling (2-3 inches in total length). However, in recent years, eggs, fish larvae (less than 1 inch) and even brood fish are also imported. Such practices contribute to the spread of fish pathogen especially viruses. Aeromonas hydrophila, the most common bacterial pathogen in fresh water fish, Aeromonas species are enteropathogens which possess virulence properties, such as, the ability to produce enterotoxins, cytotoxins, haemolysins and ability to invade epithelial cells. The main virulence factors of A.hydrophilaspecies that can be associated with gastroenteritis. Immunostimulants seems to be representing a useful alternative to vaccination and chemotherapy in the control of fish disease. They can enhance the non specific immune response. Recently in aqua culture, scores of plant extracts have been tested and used with good results in the control of bacterial and viral disease. Aloe vera is an active compound and it includes polysaccharide, mannans, anthraquinones, lectins, salicylic acid, urea, nitrogen, cinnamic acid, phenol, and sulphur. It also contains amino acids, lipids, sterols, tannin and enzymes. The world health organization (WHO) reports that over 70% of the world population needs herbal medicine for primary health care. These medicines are cheaper, non toxic biodegradable and bio compatible. There is a growing interest among researcher and feed companies for the incorporation of herbs in fish feeds as immunostimulants. Fish are an integral component of aquatic ecosystems. In addition to being a desired resource for users of the aquatic habitat, they play important role in energy flow, cycling of nutrients and maintaining community balance in the ecosystem10. Aloe vera is a perinial succulent cactus like plant, which grows in hot dry climates. Aloe vera supplementation can increase the resistance to A.hydrophila and A.Septicemia11. The relative percent survival was found to be increased in the fish fed with Aloe vera so that Aloe vera can enhance some of specific and non specific immune responses.ie lysosome activity, serum bacterial activity, total protein and Ig M level. Most of the herbs and herb extracts can be given orally which is the most convenient method of immunostimulation. This study helps to get a wider acceptance for the usage of medical plants in disease prevention and improved health management in aquaculture practices. Material and Methods Collection and maintenance of Cyprinus carpio: Fingerlings of C. carpio were collected from Manju fish farm at Kallidaikurichi, Tirunelveli district, Tamil Nadu, India. The collected fishes (10±1g) were acclimated to laboratory Research Journal of Animal, Veterinary and Fishery Sciences ________________________________________ ISSN 2320 – 6535 Vol. 2(5), 1-5, May (2014) Res. J. Animal, Veterinary and Fishery Sci. International Science Congress Association 2 conditions for 20 days in non chlorinated bore well water. Fishes were reared in the experimental tanks for 30 days in treatment with plant extract supplemented food of different concentrations. Feed for the fish: The fish feed was prepared from groundnut oil cake, wheat bran and soya bean which were purchased from the local market and were made into the fine granules mechanically. The fine granules were then, made into powder by using a mixie. The powdered groundnut oil cake, soya bean and wheat bran were mixed in the ratio of 5:2:1 and sterilized. A multivitamin tablet was also added. It was made into small pellets12. Preparation of experimental feed: The experimental fish were fed with a prepared feed as described earlier with little modification; 250,500,750 mg/kg A.vera leaf extract were mixed with sterilized dry diet and fed to healthy normal fish. After 30 days of feeding a challenge test was conducted by injecting the fish pathogen A. hydrophila, 0.1 ml of 10 CFU/ml of A. hydrophila was intraperitonally injected for control and experimental fishes. Every seven days, the Cumulative percentile mortality, oxygen consumption and opercular movement were studied. Cumulative percentile mortality: It was estimated by observing the number of fish survived throughout the experiment. The survival rate of common carp, Cyprinus carpiowas obtained by dividing the number of the fish survived to the total number of fishes. Number of fishes survived Survival rate = Total number of fishes Oxygen consumption in fish: The oxygen consumption of the fingerlings of the control and experimental fish was estimated by Winklers method. Opercular movement: The fish is taken in a beaker containing water. The number of opercular movements for a minute was recorded with the help of a stop watch in the control and the experimental fish. The triplicate observation was recorded from each sample for the control and the experimental fishes. Results and Discussion The decrease in haemoglobin concentration indicates the fish inability to provide sufficient oxygen to the tissues13. It is found that, generally mortality was not observed in 250mg/kg fish group (table-1). The control group showed higher range of (40%) mortality. No Mortality was observed in any group of fish throughout the experimental period14. The survival rate challenging the fish, Labeorohita wih A. hydrolium was enhanced when the fishes fed with the diets containing Magnifera kernel15. The relative percent survival was found to be increased in fish fed with A.vera and is able to enhance specific and non specific immune response in the C.carpio. The opercular movement of fishes in experimental group increased after the treatment (table 2). The length of the fish increased during the period and opercular beats and respiratory rate were also increased. Opercular beat, respiratory rate depend upon size of fish and stress of temperature. Similar finding was observed like a rise in atmospheric temperature due to natural variation would directly influence the water temperature and the ectothermic fishes16. The temperature and growth rate are as modulators of the metabolic capacity of fish17. The 96 hours LC50 of the household detergents Det-I is 28.5mg/litre and Det-II 41.75mg/litre in Garramullya. They also noticed changes in behavioural responses of Garamullya like opercular movement was 12-15 times more faster than controlled, frequent surfacing, loss of nervous control, try to jump out of media18. The experimental group of 250 mg/kg showed increased Oconsumption of fish (table 3). But the control group showed decrease in oxygen consumption level. The experimental fish treated with 10 mg of turmeric powder showed increase in the 2 consumption of fish, when compared to 20mg and 30 mg of turmeric powder19. Table-1 Cumulative percentage mortality (%) of fingerlings of common carp, C. carpio fed with different concentrations of Aloevera and intraperitonially challenged with 0.1ml of 10cells of Aeromonas hydrophila Dose (mg/kg) of plant extract Days after administration 0 7 14 21 28 35 Control 0 20 30 40 40 40 250 0 0 0 0 0 0 500 0 0 10 20 20 20 750 0 0 20 30 30 30 Research Journal of Animal, Veterinary and Fishery Sciences ________________________________________ ISSN 2320 – 6535 Vol. 2(5), 1-5, May (2014) Res. J. Animal, Veterinary and Fishery Sci. International Science Congress Association 3 Figure-1 Percentage mortality (%) of fingerlings of common carp, C. carpioTable-2 The opercular movement (No/ minute)of fingerlings of Common carp, C. carpio fed with different concentrations of Aloe vera and intraperitonially challenged with 0.1ml of 10CFU/ml of A.hydrophila. Each value (mean ± SD) represents the average performance of five estimations Dose(mg/kg) of plant extract Days after administration 0 7 14 21 28 35 Control 88.33 ±1.00 86.66 ±1.52 83.00 ±1.00 81.00 ±1.52 77.33 ±1.00 76.11 ±2.04 250 86.00 ±2.01 91.33 ±1.00 94.00 ±1.52 97.33 ±0.57 99.66 ±1.52 103.00 ±2.00 500 89.30 ±1.00 93.00 ±2.08 98.30 ±1.52 99.60 ±2.00 102.00 ±2.05 105.30 ±1.52 750 90.33 ±2.08 93.66 ±1.00 95.30 ±2.00 98.33 ±2.00 103.00 ±1.08 106.30 ±1.00 Figure-2 The opercular movement (No/ minute) of fingerlings of Common carp, C. carpio Research Journal of Animal, Veterinary and Fishery Sciences ________________________________________ ISSN 2320 – 6535 Vol. 2(5), 1-5, May (2014) Res. J. Animal, Veterinary and Fishery Sci. International Science Congress Association 4 Table-3 Oxygen consumption (mg/g/h) of fingerlings of common carp, C. carpio fed with different concentrations of Aloevera and intraperitonially challenged with 0.1ml of 10cells of Aeromonas hydrophila Dose(mg/kg) of plant extract Days after administration 0 7 14 21 28 35 Control 0.265 ±0.003 0.231 ±0.003 0.208 ±0.002 0.154 ±0.002 0.142 ±0.002 0.129 ±0.002 250 0.141 ±0.001 0.184 ±0.002 0.203 ±0.002 0.207 ±0.001 0.218 ±0.001 0.236 ±0.001 500 0.105 ±0.001 0.113 ±0.003 0.143 ±0.003 0.155 ±0.001 0.174 ±0.003 0.183 ±0.002 750 0.094 ±0.001 0.065 ±0.002 0.085 ±0.003 0.113 ±0.002 0.123 ±0.002 0.141 ±0.002 Figure-3 Oxygen consumption (mg/g/h) of fingerlings of common carp, C. carpioConclusionAccording to this study, the plant extract treatment of Aloveraincreased the survival rate of the fish, C.carpio. The oxygen consumption and opercular movement of the disease induced fish is low, when compared to the fish treated with the plant extract. 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