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In vivo anthelmintic activity of Butea monosperma against Trichostrongylid nematodes in sheep
Fitoterapia 77 (2006) 137 – 140 www.elsevier.com/locate/fitote
Short report
In vivo anthelmintic activity of Butea monosperma against Trichostrongylid nematodes in sheep Zafar Iqbal a,⁎, Muhammad Lateef a , Abdul Jabbar a , Muhammad Nabeel Ghayur b , Anwarul Hassan Gilani b a
b
Department of Veterinary Parasitology, University of Agriculture, Faisalabad-38040, Pakistan Department of Biological and Biochemical Sciences, The Aga Khan University Medical College, Karachi-74800, Pakistan Received 12 November 2004; accepted 17 November 2005 Available online 4 January 2006
Abstract Seeds of Butea monosperma administered as crude powder (CP) at doses of 1, 2 and 3 g/kg to sheep naturally infected with mixed species of gastrointestinal nematodes exhibited a dose and a time-dependent anthelmintic effect. The maximum reduction of 78.4% in eggs per gram of feces (EPG) was recorded on day 10 after treatment with 3 g/kg. Levamisole (7.5 mg/kg), a standard anthelmintic agent, exhibited 99.1% reduction in EPG. © 2005 Elsevier B.V. All rights reserved. Keywords: Butea monosperma; Anthelmintic; Nematodes; Sheep
1. Plant Butea monosperma (Lam.) Kuntze (Leguminosae), seeds provided from local market (Faisalabad, Pakistan), were identified and authenticated by Ass. Prof. Mansoor Hameed, Taxonomist. Samples were preserved in the Herbarium of the Department of Botany, University of Agriculture, Faisalabad. 2. Uses in traditional medicine B. monosperma is traditionally used for worms, constipation, piles, diabetes and congested throat [1]. 3. Previously isolated constituents Glycosides in leaves and flowers, alkaloids in seeds [2]. 4. Tested material Seeds shade dried, ground to powder (CP) and stored at 4 °C until use. ⁎ Corresponding author. Tel.: +92 41 9201 106 / +92 41 9200 161x3130. E-mail address: [email protected] (Z. Iqbal). 0367-326X/$ - see front matter © 2005 Elsevier B.V. All rights reserved. doi:10.1016/j.fitote.2005.11.013
138
Z. Iqbal et al. / Fitoterapia 77 (2006) 137–140
5. Animals Twenty sheep of either sex (young stock ≤ 1 year, weighing 18–24 kg) were used for the experiment conducted in the Livestock Experiment Station, Rakh Kherewala (Punjab, Pakistan). Before the start of experiment, animals were confirmed to be naturally infected with mixed species of gastrointestinal nematodes (Haemonchus contortus, Trichostrongylus colubriformis, T. axei, Oesophagostomum columbianum, Strongyloides papillosus and Trichuris
A
Eggs /g of faeces
1000
Control Levamisole (7.5 mg/kg) CP
750
*
500
** 250
0
*** 2
1
3
[Dose] g/kg
B Eggs /g of faeces
1000
*P < 0.05 **P < 0.01 ***P < 0.001
750
** 500
***
250
0
*** 1
2
3
[Dose] g/kg
C Eggs /g of faeces
1000
750
**
500
*** 250
0
*** 1
2
3
[Dose] g/kg Fig. 1. Bar diagram showing the dose-dependent (1–3 g/kg) anthelmintic activity of B. monosperma crude powder (CP) in sheep naturally infected with mixed species of gastrointestinal nematodes at (A) 5 days PT, (B) 10 days PT and (C) 14 days PT. Activity is compared with that of positive control levamisole. Values shown are mean ± SEM of four determinations; *P b 0.05, **P b 0.01, ***P b 0.001 vs. negative control.
Z. Iqbal et al. / Fitoterapia 77 (2006) 137–140
139
ovis) by qualitative and quantitative fecal examination using standard parasitological procedures [3]. Identification of nematode eggs in the feces was done using standard methods [4,5]. 6. Studied activity Twenty animals were randomly divided into five groups of four animals each and assigned to different treatments. Group 1 (control) received no treatment. Group 2 (positive control) received a single oral dose of 7.5 mg/kg levamisole HCl (Nilverm 1.5% w/v; Animal Health Division, ICI Pakistan Limited). Groups 3, 4 and 5 were treated with single doses of CP at 1, 2 and 3 g/kg, respectively. Fecal samples of each group were collected in the morning, starting from day 0 pre-treatment and at day 5, 10 and 14 post-treatment (PT) and were evaluated for the presence of worm eggs by salt floatation technique [4]. The eggs were counted by the McMaster method [3]. Egg count percent reduction (ECR) was calculated using the following formula: ECRð%Þ ¼
Pre treatment egg count per gram Post treatment egg count per gram 100 Pre treatment egg count per gram
7. Statistical analysis The data were statistically analyzed using SAS software [6]. Results were expressed as mean ± standard error of mean (SEM, n = number of experiments). The results were considered significant with P b 0.05. 8. Results and conclusions The crude powder of B. monosperma seeds (CP) showed a dose-dependent (1–3 g/kg) and a time-dependent anthelmintic activity in sheep (Fig. 1). CP showed a maximum reduction of 78.4% in eggs per gram of feces (EPG) on day 10 post-treatment which was maintained till day 14 with the dose of 3 g/kg. In comparison to this, the standard anthelmintic agent levamisole [7] exhibited 99.1% (P b 0.001) reduction in EPG with 7.5 mg/kg. The anthelmintic activity of different species of Butea has been reported against Ascaridia galli [8], Ascaris lumbricoides [9], earthworms [10,11], Toxocara canis [12], Oxyurids, Dipylidium caninum and Taenia [13]. Therefore, it seems that its use in the ethno-veterinary system of Pakistan as an anthelmintic agent is justified, particularly by the sheep farmers who do not have access to modern anthelmintics due to their non-availability or high cost. However, the dose of B. monosperma to be used for its anthelmintic effect needs to be standardized for different animal species because a dose (1.0 g/kg) that proved highly effective against A. galli in poultry and against hookworms in dogs, did not work against H. contortus, or O. columbianum, in sheep [8].
Acknowledgements This research was funded by the University of Agriculture, Faisalabad (Pakistan) under promotion of research scheme.
References [1] [2] [3] [4]
Kapoor LD. Butea monosperma. CRC handbook of Ayurvedic medicinal plants. Boca Raton: CRC Press; 1990. p. 86. Kalesaraj R, Kurup PA. Indian J Pharmacol 1962;24:63. Soulsby EJL. Helminths, arthropods and protozoa of domesticated animals. London: English Language Book Society; 1982. MAFF. Parasitological laboratory techniques, Tech Bull., No. 18. Ministry of Agriculture Fisheries and Food Manual of Veterinary. London: Her Majesty's Stationary Office, 1979. [5] Thienpont D, Rochette F, Vanparjis OFJ. J Res Foun Beerse 1979;45. [6] SAS. Statistical analysis system: user's guide. North Carolina: Statistical Institute; 1998. [7] Edwards G, Breckenridge AM. Clin Pharmacokinet 1988;15:67.
140 [8] [9] [10] [11] [12] [13]
Z. Iqbal et al. / Fitoterapia 77 (2006) 137–140 Joshi HC. Orissa Vet J 1970;5:5. Lal J, Chandra S, Sabir M. Indian J Pharmacol Sci 1978;40:97. Chandra S, Sabir M. Indian J Pharm Sci 1978;5:97. Shilaskar DV, Parashar GC. Indian J Indig Med 1989;6:49. Kalesaraj R, Kurup PA. Indian J Med Res 1968;56:1818. Narayana K, Setty DRL, Rao HS, Kamalapur PN. Mysore J Agric Sci 1976;10:98.
Short report
In vivo anthelmintic activity of Butea monosperma against Trichostrongylid nematodes in sheep Zafar Iqbal a,⁎, Muhammad Lateef a , Abdul Jabbar a , Muhammad Nabeel Ghayur b , Anwarul Hassan Gilani b a
b
Department of Veterinary Parasitology, University of Agriculture, Faisalabad-38040, Pakistan Department of Biological and Biochemical Sciences, The Aga Khan University Medical College, Karachi-74800, Pakistan Received 12 November 2004; accepted 17 November 2005 Available online 4 January 2006
Abstract Seeds of Butea monosperma administered as crude powder (CP) at doses of 1, 2 and 3 g/kg to sheep naturally infected with mixed species of gastrointestinal nematodes exhibited a dose and a time-dependent anthelmintic effect. The maximum reduction of 78.4% in eggs per gram of feces (EPG) was recorded on day 10 after treatment with 3 g/kg. Levamisole (7.5 mg/kg), a standard anthelmintic agent, exhibited 99.1% reduction in EPG. © 2005 Elsevier B.V. All rights reserved. Keywords: Butea monosperma; Anthelmintic; Nematodes; Sheep
1. Plant Butea monosperma (Lam.) Kuntze (Leguminosae), seeds provided from local market (Faisalabad, Pakistan), were identified and authenticated by Ass. Prof. Mansoor Hameed, Taxonomist. Samples were preserved in the Herbarium of the Department of Botany, University of Agriculture, Faisalabad. 2. Uses in traditional medicine B. monosperma is traditionally used for worms, constipation, piles, diabetes and congested throat [1]. 3. Previously isolated constituents Glycosides in leaves and flowers, alkaloids in seeds [2]. 4. Tested material Seeds shade dried, ground to powder (CP) and stored at 4 °C until use. ⁎ Corresponding author. Tel.: +92 41 9201 106 / +92 41 9200 161x3130. E-mail address: [email protected] (Z. Iqbal). 0367-326X/$ - see front matter © 2005 Elsevier B.V. All rights reserved. doi:10.1016/j.fitote.2005.11.013
138
Z. Iqbal et al. / Fitoterapia 77 (2006) 137–140
5. Animals Twenty sheep of either sex (young stock ≤ 1 year, weighing 18–24 kg) were used for the experiment conducted in the Livestock Experiment Station, Rakh Kherewala (Punjab, Pakistan). Before the start of experiment, animals were confirmed to be naturally infected with mixed species of gastrointestinal nematodes (Haemonchus contortus, Trichostrongylus colubriformis, T. axei, Oesophagostomum columbianum, Strongyloides papillosus and Trichuris
A
Eggs /g of faeces
1000
Control Levamisole (7.5 mg/kg) CP
750
*
500
** 250
0
*** 2
1
3
[Dose] g/kg
B Eggs /g of faeces
1000
*P < 0.05 **P < 0.01 ***P < 0.001
750
** 500
***
250
0
*** 1
2
3
[Dose] g/kg
C Eggs /g of faeces
1000
750
**
500
*** 250
0
*** 1
2
3
[Dose] g/kg Fig. 1. Bar diagram showing the dose-dependent (1–3 g/kg) anthelmintic activity of B. monosperma crude powder (CP) in sheep naturally infected with mixed species of gastrointestinal nematodes at (A) 5 days PT, (B) 10 days PT and (C) 14 days PT. Activity is compared with that of positive control levamisole. Values shown are mean ± SEM of four determinations; *P b 0.05, **P b 0.01, ***P b 0.001 vs. negative control.
Z. Iqbal et al. / Fitoterapia 77 (2006) 137–140
139
ovis) by qualitative and quantitative fecal examination using standard parasitological procedures [3]. Identification of nematode eggs in the feces was done using standard methods [4,5]. 6. Studied activity Twenty animals were randomly divided into five groups of four animals each and assigned to different treatments. Group 1 (control) received no treatment. Group 2 (positive control) received a single oral dose of 7.5 mg/kg levamisole HCl (Nilverm 1.5% w/v; Animal Health Division, ICI Pakistan Limited). Groups 3, 4 and 5 were treated with single doses of CP at 1, 2 and 3 g/kg, respectively. Fecal samples of each group were collected in the morning, starting from day 0 pre-treatment and at day 5, 10 and 14 post-treatment (PT) and were evaluated for the presence of worm eggs by salt floatation technique [4]. The eggs were counted by the McMaster method [3]. Egg count percent reduction (ECR) was calculated using the following formula: ECRð%Þ ¼
Pre treatment egg count per gram Post treatment egg count per gram 100 Pre treatment egg count per gram
7. Statistical analysis The data were statistically analyzed using SAS software [6]. Results were expressed as mean ± standard error of mean (SEM, n = number of experiments). The results were considered significant with P b 0.05. 8. Results and conclusions The crude powder of B. monosperma seeds (CP) showed a dose-dependent (1–3 g/kg) and a time-dependent anthelmintic activity in sheep (Fig. 1). CP showed a maximum reduction of 78.4% in eggs per gram of feces (EPG) on day 10 post-treatment which was maintained till day 14 with the dose of 3 g/kg. In comparison to this, the standard anthelmintic agent levamisole [7] exhibited 99.1% (P b 0.001) reduction in EPG with 7.5 mg/kg. The anthelmintic activity of different species of Butea has been reported against Ascaridia galli [8], Ascaris lumbricoides [9], earthworms [10,11], Toxocara canis [12], Oxyurids, Dipylidium caninum and Taenia [13]. Therefore, it seems that its use in the ethno-veterinary system of Pakistan as an anthelmintic agent is justified, particularly by the sheep farmers who do not have access to modern anthelmintics due to their non-availability or high cost. However, the dose of B. monosperma to be used for its anthelmintic effect needs to be standardized for different animal species because a dose (1.0 g/kg) that proved highly effective against A. galli in poultry and against hookworms in dogs, did not work against H. contortus, or O. columbianum, in sheep [8].
Acknowledgements This research was funded by the University of Agriculture, Faisalabad (Pakistan) under promotion of research scheme.
References [1] [2] [3] [4]
Kapoor LD. Butea monosperma. CRC handbook of Ayurvedic medicinal plants. Boca Raton: CRC Press; 1990. p. 86. Kalesaraj R, Kurup PA. Indian J Pharmacol 1962;24:63. Soulsby EJL. Helminths, arthropods and protozoa of domesticated animals. London: English Language Book Society; 1982. MAFF. Parasitological laboratory techniques, Tech Bull., No. 18. Ministry of Agriculture Fisheries and Food Manual of Veterinary. London: Her Majesty's Stationary Office, 1979. [5] Thienpont D, Rochette F, Vanparjis OFJ. J Res Foun Beerse 1979;45. [6] SAS. Statistical analysis system: user's guide. North Carolina: Statistical Institute; 1998. [7] Edwards G, Breckenridge AM. Clin Pharmacokinet 1988;15:67.
140 [8] [9] [10] [11] [12] [13]
Z. Iqbal et al. / Fitoterapia 77 (2006) 137–140 Joshi HC. Orissa Vet J 1970;5:5. Lal J, Chandra S, Sabir M. Indian J Pharmacol Sci 1978;40:97. Chandra S, Sabir M. Indian J Pharm Sci 1978;5:97. Shilaskar DV, Parashar GC. Indian J Indig Med 1989;6:49. Kalesaraj R, Kurup PA. Indian J Med Res 1968;56:1818. Narayana K, Setty DRL, Rao HS, Kamalapur PN. Mysore J Agric Sci 1976;10:98.