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ThesisItem Open Access Assesment of cadmium toxicity in cattle of Eloor industrial area(Department of Veterinary Pharmacology and Toxicology,College of Veterinary and Animal Sciences, Mannuthy, 2001) Nisha, A R; KAU; Chandrasekharan, A M NairA study was undertaken to assess impact of environmental pollution with cadmium in cattle of Eloor industrial belt. Areas around Fertilizers and Chemicals Travancore (FACT), India, Aluminium Company (INDAL), Binani Zinc and Hindustan Insecticides Limited (HIL) were selected for the study. As the first step, retrospective analysis of case sheets was done in Eloor and Muppathadam Veterinary hospitals. In Eloor veterinary hospital out of the 1503 case sheets examined 851 digestive disorders, 126 repeat breeders, 75 respiratory diseases and 41 deficiency diseases were observed. Out of 5920 case sheets analysed in Muppathadam veterinary hospital 3348 digestive disorders, 1284 repeat breeders 398 respiratory diseases and 433 deficiency diseases were observed. Disease conditions like digestive disorders, deficiency diseases and repeat breeders are suggestive of cadmium toxicity. The field samples like water and forages, biological samples like blood, serum, urine dung and milk were collected from cattle in the industrial field localities. The cadmium content in this field and biological sample were estimated by Atomic Absorption Spectrophotometer. Animals kept in the University Livestock Farm, Mannuthy were taken as controls. Field samples like water and fodder were also collected from area around University Livestock Farm, Mannuthy. The cadmium content of water from Alupuram, Binanipuram, Eloor south, Eloor north ranges from 0.03 – 0.04 ppm. These were significantly higher than controls (0.01 ppm). The cadmium content of fodder from Alupuram, Binanipuram, Eloor south, Eloor north ranges from 2.60 – 6.53. These were also significantly higher than controls (0.65 ppm). The cadmium level of blood, serum, milk, urine and dung from cattle of Alupuram, Binanipuram, Eloor south and Eloor north were significantly higher than controls. Haematologocal values like total erythrocyte count, haemoglobin and packed cell volume showed significant decrease in catle of test areas than controls. Other haematologic values like differential leucocyte count, total leucocyte count, mean corpuscular volume, mean corpuscular haemoglobin and mean corpuscular haemoglobin concentration did not show any significant changes. Total serum protein and albumin values from cattle of Alupuram, Binanipuram, Eloor south and Eloor north were significantly lower than control values. Serum enzymes like aspartate amino transferase, alamine amino transferase and alkaline phosphatase levels were higher in cattle from test areas than controls. It can be concluded that field and biological samples collected from the vicinities around Fertilizers and Chemicals, Travancore (FACT), Binani Zinc and Indian Aluminium Company (INDAL) are contaminated with cadmium.ThesisItem Open Access Assessment of Dichlorodiphenyltrichloroethane (DDT ) residues in cattle of floor area(Department of Pharmacology and Toxicology, College of Veterinary and Animal Sciences, Mannuthy, 2003) Deepa, A K; KAU; Usha, P T AA study was conducted to assess the impact of environmental pollution with DDT in cattle of Eloor industrial belt. Hindustan Insecticides Limited (HIL) is the major pesticide manufacturing factory of the region producing pesticides like ODT. Hence Eloor was selected as the study area. Environmental samples like sludge, water and fodder, biological samples like blood, urine, dung and milk were collected from the cattle of Eloor area. Whole blood and serum samples were collected for haematology and serum biochemistry respectively. The collected samples were analysed for ODT residues in the gas liquid chromatograph (GLC). The values obtained were compared statistically with the samples collected from University Livestock Farm, Mannuthy. Higher levels of DOT residues were obtained from the environmental and biological samples from Eloor than the corresponding samples from Mannuthy. The mean levels of total 'nOT in environmental samples of Eloor were 1.463 ± 0.555 ppm (fodder), 0.00255 ± 0.00002 ppm (water) and 35.157 ± 0.198 ppm (sludge). Biological samples from Eloor contained mean total ODT of 0.201 ± 0.123 ppm (serum), 0.023 ± 0.006 (urine), 0.910 ± 0.689 ppm (dung) and 0.058 ± 0.019 ppm (milk). Environmental samples from Mannuthy contained mean total DDT of 0.062 ± 0.021 ppm, 0.011 ± 0.006 ppm, 0.000085 ± 0.00007 ppm in fodder, sludge and water respectively. Serum, urine and dung from cattle of Mannuthy contained mean total DOT of 0.252 ± 0.145 ppm, 0.030 ± 0.003 ppm and 0.026 ± 0.017 ppm respectively. No DOT residues were detected in the milk samples from Mannuthy. The haematological values from Eloor cattle remained within the normal range except differential leucocyte count which exhibited marked neutropenia and eosinophilia. Values of total protein and albumin were normal in both groups of animals. Serum enzymes like alanine aminotransferase and aspartate aminotransferase were significantly different fr~m Mannuthy sample but were within the normal range. It can be inferred that the DDT residues present in the field and biological samples were not high enough to cause toxicity in cattle. Variation in the haematological and biochemical parameters may be due to the presence of other environmental pollutants along with DDT in the discharged industrial effluents.ThesisItem Open Access Anti-ulcer effect of Azadirachta indica (neem) and Eupatorium triplinerve (Ayyappana) in rats(Department of Pharmacology and Toxicology, College of Veterinary and Animal Sciences, Mannuthy, 2005) Sangeetha Satheesan; KAU; Joy, A DThe present study was conducted in adult albino rats to assess the anti-ulcer effect of alcoholic extract and powder of leaves of Azadirachta indica and Eupatorium triplinerve in comparison with famotidine, a standard anti-ulcer drug. Ninety six rats weighing 150-200gm body weight of either sex divided into twelve groups were used for the study with eight rats in each group. Group 1 was administered the vehicle, five per cent gum acacia for seven days where as group 2 was administered aspirin at the dose of 200mg/kg for seven days. Group 3 was administered aspirin for seven days and from 8th day onwards, they were maintained by normal feeding and watering for 20 days to assess the natural healing. Famotidine, a standard anti-ulcer drug was given at the dose of 40mg/kg for 20 days following aspirin administration to the group 4 animals. The alcoholic extract and powder of A.indica and E.triplinerve were administered to the treatment groups (Groups 5-12) at the two dose levels of 250mg/kg, 500mg/kg and 500mg/kg, 1000mg/kg respectively for 20 days following aspirin administration. The control group as well as the aspirin group were sacrificed on the 8th day, whereas all the other groups were sacrificed on 28th day. The number of ulcers and severity (ulcer score) were determined with the help of magnifying lens and the ulcer index and healing index were calculated. Various biochemical parameters were studied to confirm the anti-ulcer activity of the plant preparations under study. The degree of lipid peroxidation, as well as the anti-oxidant enzyme status, namely, superoxide dismutase and catalase levels were assessed in gastric mucosa. The serum alkaline phosphatase activity was estimated on 28th day of the experiment. Body weight was taken at weekly intervals. Heamatological parameters such as total leucocyte count, differential count and haemoglobin count were determined to assess any changes in the haemogram. Histopathological study was also conducted to evaluate the severity of ulceration and healing process. The results indicated that all the treatment groups under study produced a significant decrease in ulcer index when compared to aspirin treated control group. The alcoholic extract at the two dose levels 250mg/kg and 500mg/kg as well as powder at the two dose levels 500mg/kg and 1000mg/kg of both A.indica and E.triplinerve were found to produce a dose-dependent healing effect which is comparable to that of famotidine. Administration of the herbal formulation at the various dose levels brought about a significant reduction in lipid peroxidation and an increase in the activities of anti-oxidant enzymes namely, superoxide dismutase and catalase, which suggest its efficacy in preventing free radical induced damage. The treatment reversed the increased activity of serum alkaline phosphatase observed in the aspirin treated group. There was no significant difference in the mean body weight gain between any of the groups. Haematological study revealed no significant change and all values fall within the normal range of blood value for the species under study. The results are substantiated by the histopathological studies, which confirmed that treatment with A.indica and E.triplinerve inhibited aspirin-induced necrosis, haemorrhage and congestion in gastric mucosa. Phytochemical analysis of the extracts revealed the presence of tannins, saponins, flavonoids, triterpenes, alkaloids, phenolic compounds which are known to affect the integrity of mucous membranes. In the present study, the fact that A.indica and E.triplinerve significantly reduced ulcer index in a dose-dependent manner supports their cytoprotective effect, which may be mediated by prostaglandins and the ulcer healing effect could be attributed to its predominant effect on the mucosal defensive factors rather than offensive factors.ThesisItem Open Access Immunomodulatory effect of fractions of ethanolic extract from Emblica officinalis (Amla) fruit pulp in mice(Department of Pharmacology and Toxicology, College of Veterinary and Animal Sciences, Mannuthy, 2005) Senthil Kumar, P K; KAU; Chandrasekharan Nair, A MThe immunomodulatory activity of acetone soluble and acetone insoluble fractions of ethanolic extract of Emblica officinalis was investigated in the present study. The extracts were also qualitatively tested for the presence of various active principles. One hundred and forty four male swiss albino mice were used to assess humoral and cellular immunity by feeding the extracts at dose level of 200 mg per kg body weight for 19 days. Few animals were also administered with dexamethasone at the dose rate of 0.75 mg per kg body weight intra peritoneally for seven days before the start of the experiment to suppress the immune system and thereby to study the effect of extracts on immunosuppressed animals also. The control group received vehicle alone (five percent gum acacia). Various physiological, haematological, biochemical, enzymatic and immunological parameters like body weight, relative organ weight, total leukocyte count, differential leukocyte count, total serum, protein, serum globulin, albumin-globulin ratio, quantification of superoxide dismutase and catalase, haemagglutination test, Jerene’s plaque forming assay to assess the humoral immune response and tests like delayed type of hypersensitivity, macrophage migration index, nitroblue tetrazolium reduction test to assess the cellular immune response were performed. Both the fractions increased the body weight, spleen weight and liver weight in normal as well as in immunosuppressed animals. A significantly increased total WBC counts on days 5, 12 and 19 and the distribution of lymphocytes on days 12 and 19 were seen in normal animals. In immunosuppressed animals, non-significant increase in both total leukocyte counts and distribution of lymphocytes from day zero to nineteen was seen. Total serum protein as well as serum globulin concentration was significantly increased and albumin globulin ratio was significantly decreased in immunocompetent animals treated with both the fractions on 12th and 19th day. However, both the fractions showed only a non-significant increase in total serum protein and serum globulin in normal as well as in immunosuppressed animals, from zero day to 19th day. Significant increase was noticed in superoxide dismutase and catalase level in immunocompetent animals treated with both the fractions on 19th day. A significant stimulation of humoral immune response as indicated by an increase in antibody titre and number of antibody producing cells on both 12th and 19th day of acetone soluble and acetone insoluble fraction treated immunocompetent animals was noted. Immunosuppressed animals also showed a non-significant increase in both antibody titre and number of antibody producing cells through out the experiment. The bone marrow cellularity and foot pad swelling reaction showed a significant stimulation in immunocompetent animals on 12th and 19th day. In immunosuppressed animals also, foot pad swelling response showed a significant increase on 19th day but bone marrow cellularity was not significant. In immunocompetent animals the acetone soluble and acetone insoluble fractions showed a significant increase in macrophage migration index (MMI) and Nitro blue Tetrazolium (NBT) dye reduction test value on 12th and 19th day. However, in immunosuppressed animals both the fractions showed only a non-significant increase. The phytochemical study on both the fractions of ethanolic extract of dried Emblica officinalis fruit pulp revealed that diterpenes and triterpenes were present in acetone soluble fraction and tannins, phenolic compounds, flavonoids, glycosides, and saponins were present in acetone insoluble fraction. Thus the present study showed the higher immunostimulant activity for acetone soluble fractions in immunocompetent as well as immunosuppressed animals.ThesisItem Open Access Comparative study of the hypoglycemic effect of Azadirachta indica (Neem),Ocimum sanctum Tulsi) and Tinospora cordifolia (Chittamruthu) and their combination in diabetic rats(Department of Pharmacology and Toxicology, College of Veterinary and Animal Sciences, Mannuthy, 2004) Archana Sathyan; KAU; Chandrasekharan Nair, A MThe present study was undertaken to assess and compare the hypoglycemic effects of alcoholic extracts of Azadirachta indica and Ocimum sanctum leaves and Tinospora cordifolia vines and their combination at 200 mg/kg dose rate in alloxan diabetic rats and to compare their efficacy with standard oral hypoglycemic drug, glibenclamide. The experiment was conducted in forty eight alloxan induced diabetic rats of either sexes for a period of 37 days. Rats were divided into six groups of eight each. Group I was administered alcoholic extract of Azadirachta indica at 200mg/kg for 30 days from 7th day onwards, Group II received Ocimum sanctum alcoholic extract at the same dose rate and Group III was given Tinospora extract at 200 mg/kg. A combination of alcoholic extracts of Azadirachta, Ocimum and Tinospora was fed to Group IV at 200 mg/kg body weight. Group V received glibenclamide at 0.5 mg/animal/day for 30 days and Group VI served as diabetic control. Blood glucose level was estimated on zero day, 7th, 15th, 21st, 30th and 37th day. Body weight was taken at weekly intervals. Serum cholesterol, serum triglyceride and liver glycogen were estimated at 37th day of the experiment. All the treatment groups (I, II, III and IV) showed a gradual increase in body weight during the experimental period, but it was seen that the body weights never returned to their original weights before the commencement of the experiment. Among the treatment Groups I, II and III, Group I given Azadirachta extract had the maximum decrease in blood glucose level. Group IV which received a combination of the three drugs had a higher reduction compared to Group I, II and III. Results suggested a synergistic effect of the combination. However, the effect was comparatively lower than that of glibenclamide. Significant reduction in serum cholesterol level was seen in Group I, II and III. Highest cholesterol level was seen in Group II and the lowest serum cholesterol level was seen in Group IV. Serum triglyceride level was highest in Group II and III. Group I had a comparatively lower serum triglyceride level. Lowest triglyceride level was shown by Group IV treated with combination. Treatment with all the three drugs resulted in no significant change in the liver glycogen levels. Combination treatment also failed to produce any significant alteration in the reduced liver glycogen levels. Group I, II and III had a liver glycogen levels comparable to that of the control Group VI. From the study, it can be concluded that among the three drugs, Azadirachta has the highest hypoglycemic and hypolipidemic effect. The combination treatment produced a comparatively higher effect than that of the three drugs, suggesting a synergistic action. However, the effects of the three drugs and their combination are less when compared to that of glibenclamide and the mechanism of action of the three plants are different from that of glibenclamide.ThesisItem Open Access Comparative study of anti-inflammatory and anti-nociceptive effect of Tinospora cordifolia (Chittamruthu)and Vitex negundo Linn.(Karinochi)in rats(Department of Pharmacology and Toxicology, College of Veterinary and Animal Sciences, Mannuthy, 2004) Jerald Irwin, A; KAU; Gopakumar, NAnti-inflammatory and anti-nociceptive effect of T. cordifolia, V. negundo and the combination of two herbal agents were assessed and compared in rats. Carrageenin induced rat paw oedema and tail flick method of nociception were adopted for anti-inflammatory and anti-nociceptive screening respectively. Diclofenac potassium at the rate of 3 mg/kg was used as a standard drug for both the studies. Anti-inflammatory effect of T. cordifolia at the rate of 100 mg/kg, V. negundo at the rate of 100 mg/kg and the combination of these agents at 50 mg/kg of each were studied in rats. Test drugs at the rate of 1000 mg/kg for both T. cordifolia and V. negundo and 500 mg of each agent in combination were given for anti-nociceptive study in rats. Anti-inflammatory effect of T. cordifolia was found effective in first phase and V. negundo in the second phase of carrageenin induced inflammation. The combination of the herbal agents produced a uniform significant inhibition in both the phases. Anti-nociceptive effect was found significant for both the herbal agents and their combination, in tail flick method of nociception. This suggested a central mediated mechanism of anti-nociception by both the agents. Adrenal parameters like gland weight, ascorbic acid and cholesterol for anti-inflammatory study suggested that T. cordifolia and V. negundo had a preventive effect on the inflammation induced changes in adrenal gland. However combination of the herbal agents was found to decrease the preventive action of the individual agents against inflammation induced changes in adrenal gland. The involvement of endogenous modulatory system for the anti-nociceptive effect of T. cordifolia and V. negundo was in contrary because of the anonymous increase of adrenal ascorbic acid and decrease of adrenal cholesterol. T. cordifolia was found effective in lowering the serum cholesterol whereas combination was not that much effective. None of the treatments were found to inhibit the lipid peroxidation induced by inflammation in plasma. Combination of T. cordifolia and V. negundo showed a significant peak increase of plasma lipid peroxide level. The rise in serum level of AST and ALT in both inflammation and nociception were not inhibited by the treatments. Haematological parameters for all groups were within the normal range. However an increase in neutrophil count than lymphocyte was noticed in carrageenin induced inflammation. Both the studies showed an increase in neutrophil count without an increase in total leukocyte count for T. cordifolia. V. negundo showed an increase in total leukocyte count in anti-inflammatory study. A decrease in total leukocyte count and neutrophil count was made by the combination of T. cordifolia and V. negundo in anti-inflammatory study. However there was an increase of total leukocyte count for combination of agents in anti-nociceptive study. Total erythrocyte count and haemoglobin concentration were increased by T. cordifolia in both the studies, whereas they were normal for combination of treatment.ThesisItem Open Access Influence of Piper longum (Linn) on the anti-inflamatory and anti-nociceptive effect of diclofenac in rats(Department of Pharmacology and Toxicology, College of Veterinary and Animal Sciences, Mannuthy, 2004) Seema, S; KAU; Joy, A DThe present study was undertaken to assess the influence of P. longum on the anti-inflammatory and anti-nociceptive effects of diclofenac in rats. The experiment was conducted in two stages. In the first stage the anti-inflammatory effect was assessed by carrageenin induced paw oedema method. Four groups of eight albino rats each were used for the study. First group was kept as control which received tween 80 only. Group II served as the positive control which received diclofenac at the dose of 3 mg/kg on the seventh day. Groups III and IV were given 100 mg/kg and 200 mg/kg P. longum extract respectively for seven days, and 3 mg/kg diclofenac on the seventh day. The percentage inhibition of paw oedema was 64.26 and 62.32 for groups III and IV, while it was 67.38 for the diclofenac treated group II. No significant change was noticed in the weight of adrenal gland in groups II, III and IV as compared with that of control. The adrenal ascorbic acid and adrenal cholesterol levels were reduced in all the treatment groups (Group II, III and IV), compared with the control group (Group I). The maximum reduction was observed in the diclofenac alone treated group. In the groups treated with P. longum extract and diclofenac, the reduction in the level of adrenal ascorbic acid and adrenal cholesterol was more for the 100 mg/kg P. longum treated group. The serum AST levels were significantly higher in Group II compared with the control group. The serum ALT and cholesterol levels showed no significant variation between groups. Variation in the haematological parameters between groups was not significant. For evaluating the influence of P. longum extract on the anti-nociceptive effect of diclofenac, four groups of eight rats each were used.The results of the anti-nociceptive study also revealed a reduction in the anti-nociceptive activity of diclofenac in the groups treated with P. longum extract and diclofenac combinations compared with the diclofenac alone treated group. The change in weight of the adrenals was not significant in the groups II, III and IV compared with that of the control. The adrenal ascorbic acid and adrenal cholesterol levels were found to be depleted in all the three treatment groups (Group II, III and IV) compared with the control group (group I). The depletion was maximum in the diclofenac treated group. In the groups treated with P. longum extract and diclofenac, the reduction in the level of adrenal ascorbic acid and adrenal cholesterol was more for the 100 mg/kg P. longum treated group. Variations in serum parameters like AST and cholesterol were not significant between groups. The serum ALT levels for groups given diclofenac alone and those given the P. longum extract and diclofenac combinations were significantly increased compared with that of the control group. The ALT level was maximum in the diclofenac treated group. The haematological parameters showed no significant variation between the groups. Phytochemical study on the alcoholic extract of P. longum revealed the presence of alkaloids. The results of the study indicate that the rats given diclofenac alone showed a greater anti-inflammatory and anti-nociceptive effect than those administered P. longum extract in combination with diclofenac. This may be due to the microsomal enzyme induction which enhances the metabolism of diclofenac. Thus the experimental findings indicate that co-administration of P. longum extract decreases the anti-inflammatory and anti-noceptive effect of diclofenacThesisItem Open Access Haematological and biochemical changes in cattle of Eloor with special reference to endosulfan(Department of Pharmacology and Toxicology, College of Veterinary and Animal Sciences, Mannuthy, 2002) Jyotsana Menon; KAU; Gopakumar, NA study was undertaken to assess the impact of environmental pollution with endosulfan in cattle of Eloor industrial belt. Hindustan Insecticide Limited is the major industrial unit handling endosulfan is situated here. So this area was selected for the conduct of study. Field samples like sludge, water and fodder and biological samples like blood, dung, urine and milk of cattle in the study area were collected. The collected samples were analysed for endosulfan level by gas liquid chromatography and compared (statistically) with those of control samples collected from University Livestock Farm (ULF), Mannuthy. A higher level of endosulfan could be detected in the field and biological samples ofEloor cattle than control animals. Milk samples collected from both Eloor area and ULF, Mannuthy had no detectable level of endosulfan in them except for one sample from the study area. Fodder was suspected to be the main source of endosulfan to cattle. A significant change was noticed in lymphocyte count, neutrophil and eosinophil count of the animals in affected area. Other haematologic values like total leucocyte count, total erythrocyte count, packed cell volume and erythrocyte indices (MC V, MCH, MCHC) did not show any significant di fferences. Eventhough the biochemical parameters like the creatinine phosphokinase and BUN level showed significant difference between test and control animals, these were within the normal range. It can be inferred that the levels of endosulfan in the field and biological samples were not sufficient enough to cause toxic symptoms and the variations in haematological and biochemical parameters might be due to role of other contaminants apart from endosulfan. However it could be concluded that field and biological samples collected from vicinities of Hindustan insecticides Ltd. were contaminated with endosulfan. The higher than normal endosulfan level in all samples may be looked with caution because of its toxic nature.ThesisItem Open Access Tranquilizing property of Clitoria ternatea Linn. (Shankupushpam), Acorus calamus Linn.(Vayampu) and Vitex leucoxylon Linn.(Atta nocchi) in rats(Department of Pharmacology and Toxicology, College of Veterinary and Animal Sciences, Mannuthy, 2001) Suresh N, Nair; KAU; Venugopalan, KThe study was conducted in sixty four adult albino rats of either sex to assess tranquillizing property of alcoholic extract of whole plant of Clitoria ternatea, roots and rhizomes of Acorus calamus and leaves of Vitex leucoxylon at two dose levels of 250 mg/kg and 500 mg/kg of body weight and compare the effect to the standard tranquillizer chlorpromazine at dose rate of 7 mg/kg. The control group was given gum acacia. The dose levels of these drugs were as follows Plants Amount of Extract (mg/kg) Clitoria ternatea 250(0 I) 500(02) Acorus calamus 250(03) 500(04) Vi/ex leucoxylon 250(05) 500(06) Chlorpromazine 7(07) Control (gum acacia) 7mllkg(08) Level of tranquillization was measured using three parameters (l) depression of spontaneous motor activity, measured using actaphotometer (2) depression of forced locomotor activity measured by decrease in time of permanence in a rota-rod and (3) decrease in aggressive behaviour, measured by aggressive behaviour test score. Haematological parameters like total erythrocytic count, total leucocytic count, differential leucocytic count and haemoglobin percentage were determined to assess any change in haemogram by these drugs. The results of present study were as follows. Clitoria ternatea possess maximum tranquillizing property among the three. At 250 mg/kg it produced significant depression of spontaneous motor activity and forced locomotor activity than control but less than the standard drug. At 500 mg/kg it produced almost similar effect, as standard at certain time intervals but altogether the effect was less than the standard. Aggressive behaviour was also reduced at both dose rates. Clitoria ternatea at 500 mg/kg was more effective than Clitoria ternatea 250 mg/kg. Acorus calamus follows Clitoria lernatea in producing tranquillization. At 250 mg/kg as well as 500 mg/kg dose rates it produced significant depression of spontaneous motor activity, forced locomotor activity and aggressive behaviour. Acorus calamus at 500 mg/kg was more effective than Acorus calamus 250 mg/kg . Vitex leucoxylon possess least effect among the three in producing tranquillization. But at 250 mg/kg as well as 500 mg/kg dose rates it produced significant depression of spontaneous motor activity, forced locomotor activity and aggressive behaviour. Vitex leucoxylon at 500 mg/kg was more effective than Vitex leucoxylon 250 mg/kg. Haematological studies revealed no significant change in haemogram by any of the plant drugs. The exact mechanism of tranquillization by these plants and their possible toxicities needs further detailed study