9.0 ANTHELMINTIC STUDIES

Transcription

1 9. ANTHELMINTIC STUDIES India is an agricultural country and its natural wealth makes a substantial contribution to its economy. Worldwide about 3 million cases of helminthiasis is exist at present, either as single or as mixed infection. However since helminthic infection are usually asymptomatic and not in morbidity, they are not always regarded as a serious disease. The extent of debility and ill health caused by worms are related to the type and duration of the infection. The prevalence of helminthic infections in domestic animals in India is rather more and many infections exist for which either no treatment is available or existing remedies are not completely useful and are uncertain. Controls of helminthiasis is essential, it has not yet received the attention it deserves. 9.1 Invitro ANTHELMINTIC STUDIES The anthelmintic activities of various extracts were studied by using the method of R.G.Mali et al. the anthelmintic activity was evaluated on adult Indian earthworm, round worm and tapeworm due to its anatomical and physiological similarity with the intestinal parasites of human being. METHOD Five groups of approximately equal size earthworm, roundworm and tapeworm (consisting of six worms in each group) were released in 5ml of desired formulation. Each worm was treated with 1% gum acacia in normal saline, albendazole (25, 5, mg. /ml.) and herbal extracts in different concentration (25, 5, mg. /ml) as well as desired formulations. The time taken for paralysis was noted when the worms did not revive even in the normal saline solution. Death was concluded when the worms lost their motility followed by fading away of their body color. 15

2 FIGURE 58 FIGURE 59 EARTH WORM 151

3 Figure -6 EARTH WORM FIGURE 61 EARTH WORM 152 E 62

4 FIGURE - 62 EARTH WORM FIGURE 63 FIGURE

5 EARTH WORM TABLE - 32 ETHANOLIC EXTRACT OF PLANTS AGAINST EARTHWORM 154

6 S.No Drug Name Paralysis Time (in Minutes) Death Time (In Minutes) 1. Standard Albendazole 14.8 ± ± mg /ml 2. Standard Piperazine 35. ± ± mg/ml 3. Ethanolic solution of Dalbergia sissoo 15mg/ml 18. ± ± mg/ml 14.5 ± ± mg/ml 11.5 ± ± Ethanolic solution of Cassia fistula 15mg/ml 3mg/ml 6mg/ml 5. Ethanolic solution of Cissus quadrangularis 15mg/ml 3mg/ml 6mg/ml 6. Ethanolic solution of Clitoria ternatea 15mg/ml 3mg/ml 6mg/ml 7. Ethanolic solution of Amorphophyllus campanulatus 15mg/ml 3mg/ml 6mg/ml 8. Formulation 1 15mg/ml 3mg/ml 6mg/ml 9. Formulation 2 15mg/ml 3mg/ml 6mg/ml Mean ± SEM 47.3 ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ±

7 FIGURE 65 Comparative Anthelmintic Study on Earth Worm By Dalbergia sissoo Vs Standard Time in Minutes Death Time Paralysis Time Pip 15mg/ml Alb 15mg/ml1 Dalbergia 15mg/ml Dalbergia 3mg/ml Dalbergia 6mg/ml Drugs FIGURE 66 Compartive Anthelmintic Study On Earthworm By Cassia fistula Vs Standard Time in Minutes Death Time Paralysis Time Pip 15mg/ml Alb 15mg/ml1 Cassia 15mg/ml Cassia 3mg/ml Cassia 6mg/ml Drugs 156

8 FIGURE 67 Comparative Anthelmintic Study on Earth Worm By Cissus quadrangularis Vs Standard Time in Minutes Death Time Paralysis Time Pip 15mg/ml Alb 15mg/ml1 Cissus 15mg/ml Cissus 3mg/ml Cissus 6mg/ml Drugs 4.2 FIGURE 68 Comparative Anthelmintic Study on Earth Worm By Clitoria ternatea Vs Standard Time in Minu Death Time Paralysis Time Pip 15mg/ml Alb 15mg/ml1 Clitoria 15mg/ml Clitoria 3mg/ml Clitoria 6mg/ml Drugs 157

9 FIGURE 69 Comparative Anthelmintic Study on Earth Worm By Amorphophyllus campanulatus Vs Standard Time in Minutes Death Time Paralysis Time Pip 15mg/ml Alb 15mg/ml1 Amorphophyllus 15mg/ml Amorphophyllus 3mg/ml Amorphophyllus 6mg/ml Drugs 158

10 FIGURE 7 FIGURE

11 FIGURE 72 FIGURE 73 16

12 FIGURE 74 FIGURE -75 CLITORIA TERNATEA 161

13 FIGURE -76 AMORPHYLLUS CAMPANULATUS FIGURE

14 FIGURE

15 TABLE-33 ETHANOLIC EXTRACT OF PLANTS AGAINST ROUNDWORM S.N o Drug Name Paralysis Time (in Minutes) 1. Standard Albendazole 25mg/ml 55.6 ± mg/ml 37.7 ±.4943 mg/ml 22. ± Standard Piperazine 25mg/ml 5mg/ml mg/ml 3. Ethanolic solution of Dalbergia sissoo 25mg/ml 5mg/ml mg/ml 4. Ethanolic solution of Cassia Fistula 25mg/ml 5mg/ml mg/ml 5. Ethanolic solution of Cissus quadrangularis 25mg/ml 5mg/ml mg/ml 6. Ethanolic solution of Clitoria Ternatea 25mg/ml 5mg/ml mg/ml 7. Ethanolic solution of Amorphophyllus campanulatus 25mg/ml 5mg/ml mg/ml 8. Formulation 1 25mg/ml 5mg/ml mg/ml 9. Formulation 2 25mg/ml 5mg/ml mg/ml Mean ± SEM 12.5 ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ±.6145 Death Time (In Minutes) ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ±

16 FIGURE -79 Comparative Anthelminitic Study on Round Worm by Dalbergia sissoo Vs Standard Time in Minutes 15 Death Time Paralysis Time Pip 25mg/ml Albe 25mg/ml Dalbergia 25mg/ml Dalbergia 5mg/ml Dalbergia mg/ml Drugs FIGURE -8 Comparative Anthelminitic Study on Round Worm by Cassia fistula Vs Standard Time in Miutes Death Time Paralysis Time Pip 25mg/ml Albe 25mg/ml Cassia 25 mg/ml Cassia 5 mg/ml Cassia mg/ml Drugs 165

17 FIGURE -81 Comparative Anthelminitic Study on Round Worm by Cissus quadrangularis Vs Standard Time in Minutes Death Time Paralysis Time Pip 25mg/ml Albe 25mg/ml Cissus 25mg/ml Cissus 5 mg/ml Cissus mg/ml Drugs FIGURE -82 Comparative Anthelminitic Study on Round Worm by Clitoria ternatea Vs Standard Time in MInutes Death Time Paralysis Time Pip 25mg/ml Albe 25mg/ml Clitoria 25 mg/ml Clitoria 5 mg/ml Clitoria mg/ml Drugs 166

18 FIGURE -83 Comparative Anthelminitic Study on Round Worm by Amorphophylluscampanulatus Vs Standard Time in Minutes Death Time Paralysis Time Pip 25mg/ml Albe 25mg/ml Amorphophyllus 25mg/ml Amorphophyllus 5mg/ml Amorphophyllus mg/ml Drugs FIGURE -84 Comparative Anthelminitic Study on Round Worm by Formulation I Vs Standard Time in Minutes Death Time Paralysis Time Pip 25mg/ml Albe 25mg/ml Form. I 25mg/ml Form. I 5mg/ml Form. I mg/ml Drugs 167

19 FIGURE -85 Comparative Anthelminitic Study on Round Worm by Formulation 2 Vs Standard Time in Minutes Death Time Paralysis Time Pip 25mg/ml Albe 25mg/ml Form. 2 25mg/ml Form. 2 5mg/ml Form. 2 mg/ml Drugs 168

20 FIGURE -86 FIGURE

21 FIGURE -88 FIGURE

22 FIGURE -9 FIGURE

23 FIGURE -92 AMORPHOPHYLLUS CAMPANULATUS FIGURE

24 FIGURE

25 TABLE-34 ETHANOLIC EXTRACT OF PLANTS AGAINST TAPEWORM S.No Drug Name Paralysis Time (in Minutes) 1. Standard Albendazole 25mg/ml 8.6 ± mg/ml 72.8 ± mg/ml 6.2 ± Standard Piperazine 25mg/ml 5mg/ml mg/ml 3. Ethanolic solution of Dalbergia sissoo 25mg/ml 5mg/ml mg/ml 4. Ethanolic solution of Cassia Fistula 25mg/ml 5mg/ml mg/ml 5. Ethanolic solution of Cissus quadrangularis 25mg/ml 5mg/ml mg/ml 6. Ethanolic solution of Clitoria Ternatea 25mg/ml 5mg/ml mg/ml 7. Ethanolic solution of Amorphophyllus campanulatus 25mg/ml 5mg/ml mg/ml 8. Formulation 1 25mg/ml 5mg/ml mg/ml 9. Formulation 2 25mg/ml 5mg/ml mg/ml Mean ± SEM ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ±.8817 Death Time (In Minutes) 24.3 ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ±

26 FIGURE -95 Comparative Anthelmintic Study on Tape Worm by Dalbergia sissoo Vs Standard Time in Minutes Death Time Paralysis Time Pipe 25mg/ml Albe 25mg/ml Dalbergia 25mg/ml Dalbergia 5mg/ml Dalbergia mg/ml Drugs FIGURE -96 Comparative Anthelmintic Study on Tape Worm by Cassia fistula Vs Standard Time in Minutes Death Time Paralysis Time Pipe 25mg/ml Albe 25mg/ml Cassia 25 mg/ml Cassia 5 mg/ml Cassia mg/ml Drugs 175

27 8 FIGURE - 98 Comparative Anthelmintic Study on Tape Worm by Clitoria ternatea Vs Standard 7 6 Time in Minutes Death Time Paralysis Time Pipe 25mg/ml Albe 25mg/ml Clitoria 25 mg/ml Clitoria 5 mg/ml Clitoria mg/ml Drugs FIGURE -97 Comparative Anthelmintic Study on Tape Worm by Cissus qudrangularis Vs Standard Time in Minutes Death Time Paralysis Time Pipe 25mg/ml Albe 25mg/ml Cissus 25mg/ml Cissus 5 mg/ml Cissus mg/ml Drugs 176

28 5 FIGURE 99 Comparative Anthelmintic Study on Tape Worm by Amorphophyllus campanulatus Vs Standard T I m e i n M i n u t e s Death Time Paralysis Time Pipe 25mg/ml Albe 25mg/ml Amorphophyllus 25mg/ml Drugs Amorphophyllus 5mg/ml Amorphophyllus mg/ml FIGURE Comparative Anthelmintic Study on Tape Worm by Formulation 1 Vs Standard FIGURE -11 Time in Minutes FIGURE Pipe 25mg/ml Albe 25mg/ml Form1-25mg/mlForm1-5mg/ml Form1 - mg/ml Drugs Death Time Paralysis Time 177

29 Comparative Anthelmintic Study on Tape Worm by Formulation 2 Vs Standard Time in Minutes Death Time Paralysis Time Pipe 25mg/ml Albe 25mg/ml Form. 2-25mg/ml Form. 2-5mg/ml Form. 2 - mg/ml Drugs FIGURE

30 9.2 INVIVO ANTHELMINTIC STUDIES Animals and management This study was undertaken at Tamilnadu Veterinary and Animal Sciences University Research Centre situated in Dharmapuri. Naturally infected sheep and goats were selected and divided into seven groups of 1 animals each respectively. The animals were individually ear tagged. Group I animals were treated with Albendazole at 1 mg. /kg. and other groups were treated with Formulation I, II, III, IV, V and VI respectively. Faecal samples were collected from sheep and goat before and after the treatment with anthelmintics. It is to be expected that the eggs discharged by the majority of the helminthes find their way out along with the faeces. Hence, a critical examination of the drug passed by the animal at least on 3 successive days would indicate the causative agent. Egg Counting Technique Stoll s Method Three gram of faeces was weighed in to a test tube graduated to 45ml. The tube is then filled upto 45 ml mark with N/1 sodium hydroxide solution and 1-12 glass beads are added and shaken to get homogeneous suspension of the faecal material. The emulsion was allowed to settle till all the froth disappeared..15 ml of the emulsion was pipetted out after light shaking and spread the pipette fluid evenly on a grease free slide. All the eggs found in the samples were counted. This number multiplied by gives the number of eggs per gram of faeces. The average of two such counts should be taken. Multiplication of this number (Egg per Gram of faeces: EPG) by the total weight of the faeces passed during the day gives the total output of eggs. 179

31 G This count was made on three consecutive days to obviate all idiosyncrasies. The average EPG numbers show an indication of the worm burden, which indicate the severity of the disease. FIGURE -12 INVIVO ANTHELMINTIC STUDIES ON GOAT Goat Goat 18

32 Pretreatm ent TABLE 35 AMOUNT OF HELMINTHES PRESENT IN NATURALLY INFECTED GOAT UNDER TREATMENT Control Albendaz Formulation ole I II III IV V VI ± ± 15.9 ± ± 153. ± ± ± ± rd day ± ± ± ± ± ± ± ± th day 14.9 ± ± ± ± ± ± ± ± st day ± ± ± ± ± ± ± ±.567 Mean ± SEM 181

33 FIGURE -13 Amount of Helminthes present in the naturally infected Goat under treatment Amount of Helminthes st day Pre treatment Control Albendazole Formulation 1 Formulation 2 Formulation 3 Formulation 4 Formulation 5 Formulation 6 Drugs 182

34 FIGURE -14 COMPARATIVE STUDY OF ANTHELMINTIC ACTIVITY AMONG FORMULATIONS VS STANDARD IN NATURALLY INFECTED GOAT Amount of helminthes Pre treatment Pre treatment Pre treatment Pre treatment 21st day rd 3rd day day 21st day 21st day 21st day 21st day 21st day Control Albendazole Formulation 1 Formulation 2 Formulation 3 Formulation 4 Formulation 5 Formulation

35 Pretreatment 71.3 ±.5976 TABLE 36 AMOUNT OF EPG VALUE OF NATURALLY INFECTED GOAT UNDER TREATMENT Control Albendazo Formulation le I II III IV V VI ± ± 74. ± ± 735. ± 83.4 ± ± rd day 73.3 ± ± ± ± ± ± ± ± th day 75.2 ± ± ± ± ± ± ± ± st day 721. ± ± ± ± ± ± ± ±.6671 Percentage Efficacy 92.31% 91.39% 9.54% 87.58% 85.71% 89.15% 79.4% Mean ± SEM 184

36 FIGURE-15 AMOUNT OF EPG VALUE OF NATURALLY INFECTED GOAT UNDER TREATMENT Amount of Egg Per Gram st day Pretreatment Control Albendazole Formulation I Formulation II Formulation III Formulation IV Formulation V Formulation VI 185

37 FIGURE-16 Comapartive study of Formulations Vs Standard based on Egg Per Gram Pretreatment Pretreatment Pretreatment Pretreatment Pretreatment 21st day Amout of Egg Per Gram rd 3rd day day 21st day 21st day 21st day 21st day 21st day Control Albendazole Formulation I Formulation II Formulation III Formulation IV Formulation V Formulation VI

38 FIGURE-17 Anthelmintic Efficacy of Formulations & Standard in Goat Percentage Efficacy Percentage Efficacy 75 7 Control Albendazole Formulation I Formulation II Formulation III Formulation IV Formulation V Formulation VI 187

39 FIGURE-18 INVIVO ANTHELMINTIC STUDIES ON SHEEP Sheep Sheep 188

40 TABLE 37 AMOUNT OF HELMINTHES PRESENT IN NATURALLY INFECTED SHEEP UNDER TREATMENT Pretreatment ±.7277 Control Albendazole Formulation I II III IV V VI ± ± ± ± ± ± ± rd day ± ± ± ± ± ± ± ± th day ± st day ±.7722 Mean ± SEM 26.4 ± ± ± ± ± ± ± ± ± ± ± ± ± ±

41 FIGURE-19 Amount of Helminthes present in the naturally infected sheep under treatment Amount of helminthes st day Pre treatment Control Albendazole Formulation 1 Formulation 2 Formulation 3 Formulation 4 Formulation 5 Formulation 6 19

42 FIGURE-11 COMPARATIVE STUDY OF ANTHELMINTIC ACTIVITY AMONG FORMULATIONS VS STANDARD IN NATURALLY INFECTED SHEEP Amount of Helminthes Pre treatment Pre treatment Pre treatment Pre treatment Pre treatment 21st day st day Control Albendazole Formulation 1 Formulation 2 Formulation 3 Formulation 4 Formulation 5 Formulation st day 21st 21st day day

43 TABLE -38 AMOUNT OF EPG VALUE OF NATURALLY INFECTED SHEEP UNDER TREATMENT Control Pretreatment 64. ±.766 Albenda zole 67.4 ±.6186 Formulation I II III IV V VI ± ± 68.5 ± ± 62.1 ± ± rd day 62. ± ± ± ± ± ± ± ± th day 645. ± ± ± ± ± ± ± ± st day 635. ± ± ± ± ± ± ± ±.768 Percentage Efficacy 84.32% 81.2% 81.95% 64.71% 76.11% 81.45% 77.52% Mean ± SEM 192

44 FIGURE-111 AMOUNT OF EPG VALUE OF NATURALLY INFECTED SHEEP UNDER TREATMENT Amount of Egg Per Gram st day Pretreatment Control Albendazole FormulationI FormulationII FormulationIII Formulation IV FormulationV Formulation VI 193

45 FIGURE-112 Comparative study of Formulations Vs Standard based on Egg Per Gram in sheep Amount of Egg Per Gram Pretreatment Pretreatment Pretreatment Pretreatment Pretreatment 21st day rd 3rd day day 21st day Control Albendazole FormulationI FormulationII FormulationIII Formulation IV FormulationV Formulation VI 21st day 21st day 21st 21st day day

46 FIGURE-113 Anthelmintic Efficacy of Formulations & Standard in Sheep Percentage Efficacy FormulationI FormulationII FormulationIII Formulation IV FormulationV Formulation VI Albendazole Control Percentage Efficacy

47 FIGURE-114 EGG OBSERVED IN FECRT 196