doi:10.5455/vetworld.2013.396-399 Estimation of humoral immune response in rabbits fed with Cucurbita maxima seeds

Aim : The objective of the study was to estimate the humoral immune response in rabbits treated with Cucurbita maxima seeds. Materials and Methods: Thirty six male Newzealand White rabbits were divided into six groups (I, II, III, IV, V, VI) of six in each. Group I was the untreated control. Group II was treated with dexamethasone sodium (2 mg/Kg, i.m) for 7 days. Group III was treated with levamisole hydrochloride at 2.5 mg/kg (s.c) thrice a week. Group IV was treated with Cucurbita maxima seeds. Group V was treated with levamisole and dexamethasone and Group VI was treated with dexamethasone and Cucurbita maxima seeds. The seed was given @ 1000 mg/kg orally for 10 days. Antibody titre and serum immunoglobulin concentration were estimated along with haematology. Results: Dexamethasone caused significant decreases in the antibody titre, immunoglobulin concentration where as Curcurbita maxima, Dexamethasone + Curcurbita maxima and dexamethasone + levamisole groups showed significant increase in these entities. There were no significant differences in RBC count, Haemoglobin contents among all the groups studied.


Introduction
response of Cucurbita maxima seeds may benefit human and animal population in case of infectious Several herbs have been used traditionally to conditions.The present study was undertaken to find prevent and treat human and animal diseases.Recently, out the effect of Cucurbita maxima seeds on the scientific evaluation of plants and preparations of plant humoral immune response in rabbits.origin medications have received more attention [1].One of these plants is Cucurbita maxima, popularly

Materials and Methods
known as pumpkin is cultivated throughout the world Thirty six male Newzealand White rabbits with for use as vegetable as well as medicine.It has been the bodyweight of 1000-1500g were divided into six used traditionally as medicine in many countries such groups (I, II, III, IV, V, VI) of six in each.Group I was as India, China, Argentina, Brazil and Iran [2].The the untreated control.Group II was treated with seeds of Cucurbita maxima contain necessary fatty dexamethasone sodium (Immunosuppressive).Group acids, vitamin A and C, Calcium and Zinc [3].Pumpkin III was treated with levamisole hydrochloride has been tried successfully for its antioxidant, anti-(Immuno stimulant).Group IV was treated with malarial, antitumour, anti-inflammatory, antibacterial, Cucurbita maxima seeds (shade dried crude powder anti fungal cardioprotective and anthelmintic preparation).Group V was treated with levamisole and properties [4][5][6][7][8][9][10][11].Pumpkin seed has been used in dexamethasone and Group VI was treated with traditional medicine in North America and Mexico dexamethasone and Cucurbita maxima seeds.since long as an antihelmintic agent and for supportive The rationale behind the use of Cucurbita maxima treatment in functional disorders of the bladder and for with dexamethasone was to study the influence of difficulties in urination [12,13].Stimulation of the Cucurbita maxima seed on dexamethasone induced immune system is highly desirable for the treatment of immunosuppression status of rabbits [1,15].For immunodeficiency and infectious diseases [14].The comparison, dexamethasone was given with levamisole anti-inflammatory and cardio protective effects of (Group IV) as the latter is known to restore corticosteroid pumpkin may relate to an influence of plant compounds induced depletion of Lymphocytes.Levamisole Hcl on immunocompetent cells.The evaluation of immune was given at 2.5 mg/kg subcutaneously thrice a week, dexamethasone sodium was given at 2mg/kg intra muscularly for seven days and Cucurbita maxima was given @ 1000 mg/kg orally for 10 days.ions completely.The contents of the cellophane tube All the six groups of rabbits were immunized with were dissolved in 5 ml of normal saline.The globulin typhoid-H antigen at the dose of one ml subcutaneously concentration was measured spectrophotometrically st th and the O.D. values obtained were multiplied by 0.70.on 1 day and 15 day.Blood was collected and serum separated on day 0,7,14 and 21 for the estimation of Total erythrocyte count (RBC): It was done by Hayem's antibody titre and total serum immunoglobuline levels.dilution method.Haemoglobulin (Hb) estimation was For estimation of anibody titre, widal tube done by Sahil's acid haematin method.Total leukocyte agglutination test was performed using widal test kits count (WBC) was done by standard dilution method.(Span Diagnostics Ltd, Surat, India).
Statistical analysis: The data obtained were statistically Estimation of total serum immunoglobulin: It was analysed as per method of Snedecor, G.W. and carried out by ammonium sulphate precipitation and Cochran,W.G. [17].dialysis method [16].5 ml of saturated ammonium

Results
sulphate solution was added slowly to 5 ml serum in a test tube to get half saturation of ammonium sulphate in Dexamethasone caused significant decrease in the sample.The contents were mixed continuously by the antibody titre (14.17 ± 2.17) when compared to all swirling the tube without agitating excessively.The other groups.Groups treated with Curcurbita maxima, mixture was allowed to stand for 5 minutes so that Dexamethasone + Curcurbita maxima and dexamethaglobulin precipitates.The material was centrifuged for sone + levamisole groups showed significant increase 10 minutes at high speed.The precipitate was allowed in antibody titre (153.33 ± 23.09, 170 ± 29.08, 183.33 ± to settle down at the bottom of the tube.The 29.02, respectively) and the results were comparable supernatant fluid (containing albumin) was poured off among each other (Table-1).and the precipitate was redissolved in 1 ml distilled The dexamethasone treated group showed water.The dissolved precipitate was put in a semi significant decrease in total serum immunoglobulin permeable cellophane tube, and each end of tube was level when compared to all other groups.Groups   The results of the study suggest that Cucurbita significant increase in the total serum immunoglobulin maxima seed powder has the ability to modulate levels (41.40 ± 0.21 mg/ml, 41.43 ± 0.15 mg/ml, 40.78 humoral immune response in normal and immuno-± 0.13 mg/ml respectively) (Table-2).There were no suppressed rabbits.significant differences in total erythrocyte count, This study is a part of M.V.Sc thesis submitted by In the present study, dexamethasone produced a the first author to Tamil Nadu Veterinary and Animal significant reduction in antibody titre when compared Sciences University (TANUVAS).The author is to control which is in accordance with the findings of thankful to TANUVAS for providing the facilities to Pruett et al [18].Levamisole produced significant carry out the study.increase in titre when compared to control and was able

Competing interests
to overcome the dexamethasone suppressed antibody titre.Similar results were obtained by Babiuk and Authors declare that they have no competing interest.Mishra [19].Cucurbita maxima was able to produce a globulin level in immune suppressed animals and the

Table - 2
. Effect of immunomodulators on total serum immunoglobulin levels (Mean ± SEM) mg/ml