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IMMUNOSTIMULATION OF FISHES BY HERBAL FEED INGREDIENTS


Rao, V. Y., Singh, M. R., Singh, A. and Chakrabarti, R. 2004. Potentiation of antibody production in Indian major carp Labeo rohita, rohu by Achyranthes aspera as a herbal feed ingredients. Aquaculture 238: 67-73

Labeo rohita, rohu (20017 g), were fed with two types of diets; an experimental diet, containing root extract (0.5%) of Achyranthes aspera (Amaranthaceae) as an ingredient and control diet without the root extract. After 4 weeks of feeding, fish were immunized with chicken RBC. Antigen-specific antibody response, total serum globulin and RNA/DNA ratio of spleen were determined for 4 weeks after immunization. Antigen-specific antibody and total serum globulin levels peaked on day 14 after immunization and gradually decreased towards day 28. Though hemagglutination antibody (HA) titers were always higher in the test group than the control group, total serum globulin level was significantly higher only on days 14 and 21. The sequential relationship between RNA/DNA ratio and protein level was found in the present study, as the RNA/DNA ratio was maximum on day 7 followed by higher serum protein level on day 14 in both groups. The RNA/DNA ratio was significantly (P<0.05) higher in test group of fish than the control group on days 7 and 14. These results showed the immunostimulatory activity of the prepared diet containing root extract of A. aspera.

 

Rao, V. Y. and Chakrabarti, R. 2004. Enhanced anti-proteases in Labeo rohita fed with diet containing herbal ingredients. Indian Journal of Clinical Biochemistry 19: 132-134.

Aqueous root extract of Achyranthes aspera was incorporated in the experimental diet of Labeo rohita (rohu). Control diet was prepared without root extract. Feeding of fishes with experimental diet has significantly (P<0.05) enhanced the serum anti-proteases level than fishes fed with control diet.

 

Rao, V. Y. and Chakrabarti, R. 2005. Stimulation of immunity in Indian major carp Catla catla with herbal feed ingredients. Fish & Shellfish Immunology 18: 327-334.

Catla catla, catla (150 20 g) were fed a diet containing seed of Achyranthes aspera (0.5%) and control diet without A. aspera for four weeks prior to and after ip injection with chicken erythrocytes. Fish were sampled for four consecutive weeks after immunization. Hemagglutination antibody titers were significantly higher in the test group of fishes compared with the control group. Serum globulin levels were significantly (Pt-test<0.05) higher in the test group than control group on days 14 and 21. Anti-trypsin activity due to total serum protease inhibitors and a1-antiprotease was also significantly (Pt-test<0.05) higher in the test group of fishes than the control. RNA/DNA ratio of spleen and kidney was also significantly (Pt-test<0.05) higher in test group than the control group. All these results confirm that A. aspera enhances the immunity of catla.

 

Vasudeva Rao, Y. and Chakrabarti, R. 2005. Dietary incorporation of Achyranthes aspera seed influences the immunity of common carp Cyprinus carpio. Indian Journal of Animal Sciences 75: 1097-1102.

Achyranthes aspera seed (0.5%) was incorporated in the diet for Cyprinus carpio (9017 g); control diet was prepared without the seed of Achyranthes. After four weeks of feeding, fish were immunized with chicken RBC and were sampled for four weeks after immunization. Antigen-specific antibody response, total serum protein, serum albumin and globulin, lysozyme, serum a1-protease inhibitor and a2-macroglobulin, and RNA/DNA ratio of spleen and kidney were determined. The immune response of the fish was enhanced when fed with experimental diet containing Achyranthes aspera. Hemagglutinating antibody titres were significantly (P<0.05) higher in Achyranthes treated group than control group. Serum globulin level was significantly (P<0.01) higher in the test group than the control one on second week. Serum a2-macroglobulin level was also significantly higher in test group than control, but there was no significant difference in a1-protease inhibitor level. RNA/DNA ratio was higher in the test group than the control group. All these results confirm that Achyranthes aspera enhances the immunity of Cyprinus carpio.

 

Vasudeva Rao, Y., Das, B.K., Jyotyrmayee, P. and Chakrabarti R. 2005 Effect of Achyranthes aspera on the immunity and survival of Labeo rohita infected with Aeromonas hydrophila. Fish & Shellfish Immunology 20: 263-273 
 

Achyranthes aspera seed was incorporated in the diets (at 0.01%, 0.1% and 0.5%) of Labeo rohita, rohu fingerlings (3.0  0.4 g). After 2 weeks, the fish were immunized with heat-killed Aeromonas hydrophila, and after a further 2 weeks the rohu were experimentally infected with Aeromonas hydrophila (ATCC 49140). After 7 days blood and serum were sampled to determine superoxide anion production, bactericidal activity, lysozyme, serum protein, albumin, globulin, serum glutamate oxaloacetate transaminase (SGOT), serum glutamate pyruvate transaminase (SGPT) and alkaline phosphatase (ALP). Superoxide anion production, serum bactericidal activity, lysozyme, ALP, serum protein, albumin:globulin ratio (A/G) were enhanced in Achyranthes treated groups compared to the control group. SGOT and SGPT levels were elevated in control group, but in Achyranthes treated groups the levels were similar to the uninfected-control group. Higher cumulative mortalities were observed in the control group (77%) up to day-9 after infection. This gradually decreased with increasing dose of Achyranthes, 66% mortality in 0.01% group, 57% mortality in 0.1% group and 28% mortality in 0.5% group. These results indicate that Achyranthes aspera stimulates immunity and increases resistance to infection in L. rohita.
 

Chakrabarti, R. and Vasudeva Rao. Y. 2006. Achyranthes aspera stimulates the immunity and enhances the antigen clearance in Catla catla. International Immunopharmacology 6/5: 782-790.

Achyranthes aspera, an Indian medicinal plant (family: Amaranthaceae) was incorporated in artificial fish diet, and fed to catla Catla catla. After 4 weeks of feeding, fish were immunized with bovine serum albumin (BSA), spleen and blood were sampled on weekly intervals for four times after immunization. Antigen-specific antibody level in serum was determined by ELISA. Antigen clearance was determined in spleen by immuno-electron microscopy. Achyranthes has significantly (P<0.05) enhanced the BSA-specific antibody titers than the untreated control group throughout the study period. The efficiency of antigen clearance was also enhanced in C. catla treated with Achyranthes.

Chakrabarti, R. and Rao, V. Y. 2012. Achyranthes aspera enhances the immunity and antigen clearance in common carp Cyprinus carpio. Journal of Fish Diseases (Wiley-Blackwell) 35: 389-392.

Achyranthes aspera, an Indian medicinal plant (Family: Amaranthaceae), was incorporated in the artificial diet of common carp Cyprinus carpio L (9017 g). Control group was fed with normal diet without Achyranthes aspera. After 4 weeks of feeding, fish were intraperitoneally immunized with bovine serum albumin. Blood samples were collected from both treated and control groups at weekly intervals after immunization; and spleen was sampled on second week to study the antigen clearance property of plant ingredient by immuno-electron microscopy. Antibody response specific to BSA was determined by ELISA. Achyranthes aspera significantly (P<0.05) enhanced the antigen-specific antibody response in Cyprinus carpio. Injected antigen, i. e. BSA was efficiently cleared from the spleen of treated fish.

Srivastava, P. K. and Chakrabarti, R. 2012. Effect of dietary supplementation of seed of Achyranthes aspera on the immune system of Labeo rohita fry. Israeli Journal of Aquaculture - Bamidgeh (IJA: 64.2012.786).

Labeo rohita (rohu) fry (0.547 0.01 g) were fed one of four diets containing 0 (control), 0.1, 0.5, or 1.0% Achyranthes aspera seed. After 30 days, the fry were immunized with chicken red blood cells (c-RBC). Blood samples were collected 7, 14, and 21 days after immunization. Significantly (P < 0.05) higher average weight (2.565 0.02 g) and SGR were obtained in fry fed the 1.0% diet compared to others. The increase in average weight was directly related to the increasing dose of seed. FCR was significantly (P < 0.05) lower in fry fed the 1.0% diet while total serum protein, albumin, and globulin were higher in treated groups than in the unsupplemented control. The antigen-specific antibody titer level was significantly (P < 0.05) lower in rohu fed the control diet than in rohu fed the supplemented diets. Titer levels were 32 - 128, 128 - 256, 256 - 1024, and 256 - 1024 in the 0, 0.1, 0.5, and 1.0% diets, respectively. Serum glutamic oxaloacetic transaminase (SGOT), serum glutamate pyruvate transaminase (SGPT), and alkaline phosphatase (ALP) were significantly (P < 0.05) higher in rohu fed the control diet than in rohu fed the supplemented diets while myeloperoxidas was lower in the former. Among the treated groups, myeloperoxidase was significantly (P < 0.05) higher in rohu fed the 1.0% diet (1.561-2.558, ? 450 nm) than in those fed other diets. The present study documents the immunostimulatory properties of A. aspera seed and finds that a dose of 1.0% might be suitable for rohu fry.

Srivastava, P. K. and Chakrabarti, R. 2012. Effect of dietary supplementation of Achyranthes aspera seed on Labeo rohita larvae challenged with Aeromonas hydrophila. Journal of Aquatic Animal Health (Taylor & Francis, DOI:10.1080/08997659.2012.694834).

Labeo rohita larvae were fed with four different diets containing Achyranthes aspera seed at three different levels of 0.1 (D1), 0.25 (D2) and 0.5% (D3) and control diet (D4). After 70 days, fish were injected intraperitoneally with live Aeromonas hydrophila. Mortality of fish was recorded for 7 days. In D4, first mortality was recorded within 12 h of exposure, whereas in all treated groups, it was recorded after 24 h. In D4, 50% fish died within 72 h, whereas in D3, 10 to 15% mortality was found between 72 and 84 h. The cumulative mortality rates were 50, 40, 35 and 15% in D4, D1, D2 and D3, respectively. Total tissue protein level was higher in 0.25 and 0.5% seed supplemented diets fed larvae compared to others. Glutamic oxaloacetic transminase, glutamate pyruvate transminase and thiobarbituric acid reactive substance levels were significantly (P < 0.05) lower in 0.5% seed supplemented diet fed larvae, whereas lysozyme and nitric oxide synthase levels were significantly (P < 0.05) higher in this group compared to others. Dietary supplementation of Achyranthes aspera seed at 0.5% level provided protection against oxidative stress, prevented tissue damage and enhanced disease resistance of carp larvae.

Chakrabarti, R., Srivastava, P. K., Kundu, K., Khare, R.S. and Banerjee, S. 2012. Evaluation of immunostimulatory and growth promoting effect of seed fractions of Achyranthes aspera in common carp Cyprinus carpio and identification of active constituents. Fish & Shellfish Immunology (Elsevier B.V.) 32: 839-843.

AImmunostimulatory and growth promoting properties of Achyranthes aspera seeds were studied with larvae of common carp Cyprinus carpio. Four experimental diets were prepared using raw (D1) and alcohol (D2), petroleum ether (D3) and 50% aqueous alcohol (D4) extracts of A. aspera seeds. Diet without seed served as control (D5). Fish were fed with test/control diet for 30 days and then immunized with 10 ml of c-RBC. Blood samples were collected 7 days after immunization. Survival (93 3%) of fish was significantly (P < 0.05) higher in D1 diet fed group compared to others. Highest specific growth rate was found in fish fed with diet D2. Significantly (P < 0.05) higher levels of serum protein and albumin were found in D1 and D3 compared to others. Highest serum globulin level was found in D1, which was followed by D3, D2, D4 and D5. Hemagglutination titer level was 5 - 18 folds higher in diet D3 fed fish compared to others. SGOT and SGPT levels were significantly (P < 0.05) higher in control group compared to the treated groups. Myeloperoxidase activity was significantly (P < 0.05) higher in D1 (2.513 0.27 l 450 nm) and D3 (2.38 0.07 ? 450 nm) diets fed groups compared to others. The best performance of fish was found in raw A. aspera seeds incorporated diet fed group and the active constituents were identified as ecdysterone and two essential fatty acids linolenic acid and oleic acid.

Chakrabarti, R., Srivastava, P., Verma, N. and Sharma, J. G. 2014. Effect of seeds of Achyranthes aspera on the immune responses and expression of some immune-related genes in carp Catla catla. Fish & Shellfish Immunology (Elsevier B.V., DOI: 10.1016/j.fsi.2014.03.007).

Evaluation of functional mechanism of immunostimulant in fish is essential for realizing their therapeutic potential. The impact of dietary supplementation of seeds of Achyranthes aspera on the immune system of Catla catla was studied. Catla (37.736.7 g) were fed with test diets containing 0.5 (D1) and 1.0% (D2) seeds of A. aspera and control diet (D3) for 50 days. Then fish were immunized with heat-killed Aeromonas hydrophila. Blood and tissue samples were collected after 7 days of immunization. Three fish were sacrificed for each parameter. Respiratory burst activity, bactericidal activity and antibody response were significantly (P < 0.05) higher in test diets fed catla compared to the control one. Nitric oxide synthase and serum lysozyme were significantly (P < 0.05) higher in fish fed with diet D2 compared to the other treatments. There were up-regulations of TNFa, lysozyme c and g gene expressions in kidney and IL-10, lysozyme c and g expressions in hepatopancreas of test diets fed fish compared to the control one. The expression of IL-10 was down-regulated in the kidney of D2 diet fed fish compared to others. In the gill, significantly (P < 0.05) higher expressions of TNFa was found in D1 diet feed fish and lysozyme c and in D2 diet fed fish compared to the other groups. This study showed tissue-specific gene expression pattern in catla. An inverse relationship was found between the expression of IL-10 in kidney of catla.


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