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Int. J. BioRes. 2(11): 29-33 November, 2010 Biswas et al. Full Length Research Paper

EFFICACY OF HERBAL AND COMMERCIALLY AVAILABLE ANTIPROTOZOAL
DRUGS ON PIGEON TRICHOMONIASIS
*P G Biswas, N Begum, M A A Mamun, M A Bari and M M H Mondal
Department of Parasitology, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh ABSTRACT
Efficacy of four herbal and five commercially antiprotozoal drugs was evaluated in vitro and in vivo on pigeon trichomoniasis (causal agent Trichomonas gallinae). Experimental groups were treated with these medicines at different concentration (%) and doses and control groups were not treated with any drug. Among the herbal drugs (thankuni, neem, bishkatali and guava), thankuni 20% conc. showed the highest efficacy (80% & 75% in in vitro and in vivo condition, respectively) whereas guava (10% conc.) had the lowest efficacy (38.46% and 33.33% in in vitro and in vivo condition, respectively). Among the commercial drugs (metronidazole, tinidazole, ornidazole, secnidazole, nitazoxanide), metronidazole, ornidazole & secnidazole (30mg/kg bwt.) had the highest efficacy (100%) in both in vitro and in vivo condition whereas nitazoxanide (30mg/kg bwt.) had the lowest efficacy (84.61% and 83.33% in in vitro and in vivo condition, respectively). In both herbal and commercial drugs, efficacy was highest when higher concentration and doses were used. No exception was recorded. The present study suggests that metronidazole, ornidazole & secnidazole are the most effective drugs tested against pigeon trichomoniasis and metronidazole, also proved cost effective. Key words: Efficacy, antiprotozoal drugs, trichomoniasis, pigeon
INTRODUCTION
Trichomonas gallinae is the causative agent of trichomoniasis, a parasitic disease that mainly affects columbiforms and birds of prey, but on occasions also galliform, anseriform, passeriform and psittacine species (Stabler, 1954; Baker, 1986; Cole, 1999). It is the causative agent of canker in pigeons and causes a variety of pathologic manifestations depending on the strain of the parasite and species of bird it infect (Baker, 1986; Cooper and Petty, 1988). Pathogenic strains cause caseous, fibronecrotic lesions in the oral cavity, pharynx and upper digestive tract that generally lead to the death of the affected bird due to starvation or secondary bacterial infections (Stabler, 1954; Mesa et al., 1961; Ho fle et al., 2000; Real et al., 2000). Pigeons are also susceptible to the spread of certain pathogenic strains of T. gallinae to internal organs such as the liver (Narcisi et al., 1991). In young the infection is severe and even fatal. Mortality can reach up to 80-90% or more in squabs (Soulsby, 1982). An overall prevalence of these protozoa has been recorded 32% from Italy (Catelli et al., 1999), 11% from Chile (Toro et al., 1999). In Bangladesh, Begum et al. (2008) reported 67.33% prevalence of trichomoniasis in pigeon. An in-depth study regarding the effect of herbal and commercially available antiprotozoal drugs on pigeon trichomoniasis due to Trichomonas gallinae is required to prevent the mortality and morbidity. In Turkey Aydn et al. (2000) described 100% and 87.5% efficacy rates for metronidazole and carnidazole, respectively against Trichomonas gallinae in naturally infected pigeons. In China Luo et al. (2006) tested efficacy of common antitrichomonal drugs against Trichomonas gallinae from pigeon in vitro. Munoz et al. (1998) tested the in vivo and in vitro sensitivity of Trichomonas gallinae to some nitromidazole drugs in Spain. But in Bangladesh the efficacies of both herbal and commercially available antiprotozoal drugs have yet been established for trichomoniasis in pigeon. For this regard, the proposed investigation has been taken to address properly the in vitro and in vivo efficacy of both herbal and commercially available drugs on pigeon trichomoniasis. MATERIALS AND METHODS
The study was conducted from July 2008 to June 2009. In this study, efficacy of five commercially available drugs, namely,
metronidazole, tinidazole, ornidazole, secnidazole, nitazoxanide and fresh leaves of four herbal plants, namely, Thankuni
(Centella asiatica), Neem (Azadirachta indica), Bishkatali (Polygonum hydropiper), Guava (Psidium guajava) were
evaluated. For this, the research work was conducted in three phases: firstly, culture of the Trichomonas gallinae (sample);
secondly, in vitro drug testing and lastly, in vivo drug testing.
Culture of Trichomonas gallinae (sample)
In this study, the observations were made on the samples collected from the upper digestive tract of domestic pigeons (Columba livia). The throats of pigeons were swabbed and swabbing was inoculated into glucose-broth-serum medium (Richardson & Kendall, 1963). Antibiotic (penicillin 5,000 units and dihydrostreptomycin 1,000 microgram) was added to avoid contamination and the samples (Trichomonas gallinae) were incubated at 37°C for 3 to 6 hours. Generation time of Trichomonas gallinae was recorded as 3.30 hours to 5.50 hours (Soulsby, 1982). Int. J. BioRes. 2(11): 29-33 November, 2010 Biswas et al. In vitro drug testing
The efficacy of nine common antitrichomonal drugs (five commercial & four herbal) against Trichomonas gallinae were evaluated in vitro. All the cultured samples in which Trichomonas gallinae stayed in breeding stable phase were allocated to experimental groups and control groups randomly. In experimental groups, these drugs were given at different concentration (%) and doses, respectively and no drugs were used in control group. In this study, 10% and 20% concentration of herbal plants were used and the used dose rates of commercial drugs were 30mg/kg bwt and 20mg/kg bwt. After cultivation for 2 to 6 hours at 37˚C, the number of survived parasites (Trichomonas gallinae) were counted and compared with the control groups and relative survival rate were used to evaluate the efficacy. In vivo drug testing
About 240 Trichomonas gallinae infection free domestic pigeons (Columba livia) were collected from different local markets
of Mymensingh confirmed by microscopic examination of crop contents. In this study, these birds were grouped into 20 equal
groups, each group contained 12 birds. Experimental infection was given in 18 groups and 2 groups remained as control. After
5 days of infection, 18 groups were treated with 9 different drugs (herbal and commercial) in different concentration (%) &
doses. In this study, 10% and 20% concentration of herbal plants were used and the used dose rates of commercial drugs were
30mg/kg bwt and 20mg/kg bwt. Treatment was continued at five consecutive days with (metronidazole, tinidazole &
nitazoxanide) and single dose treatment was given with ornidazole and secnidazole. Observation was made on the samples
collected from the upper digestive tract (crop) of the pigeons and examined under microscope, from first day of treatment to
7th day after finishing of treatment. Comparison was made between the control groups & those of treated groups.
Statistical analysis
The data was analyzed statistically by using x2-test (Comparison of several proportion) (Mostafa, 1989). RESULTS AND DISCUSSION
Herbal drugs
Among the herbal plants, fresh juice of thankuni showed the highest efficacy (80%) at 20% concentration in in vitro condition whereas efficacy was 75% at same concentration in in vivo condition. This result could not be compared due to paucity of relevant literature. However, the plant contains sitosterol and tannin which have antiprotozoal activity (Daniel, 2005). Fresh juice of neem showed 71.42% efficacy at 20% concentration against Trichomonas gallinae, in vitro condition. On the other hand, in in vivo condition neem showed 66.66% efficacy at same concentration. This result could not be compared and contrasted due to lack of relevant literature. Neem has several medicinal uses. Neem leaves contains flavonid, quercetin, nimbiodol and tannins (Ghani, 2003). Flavonid, quercetin, nimbiodol (Khalid et al., 1986) and tannin (Daniel, 2005) have antiprotozoal activity. Besides this, neem leaves contain vitamin C, which also act as antioxidant (Kayser et al, 2002). Table 1: Efficacy of herbal drugs against Trichomonas gallinae in in vitro condition χ2 value
* Means significant at 1% level; #Pre -treated live T. galline counted; ^Post -treated live T.galline counted Table 2: Efficacy of herbal drugs against Trichomonas gallinae in in vivo condition χ2 value
Int. J. BioRes. 2(11): 29-33 November, 2010 Biswas et al. It was observed that, fresh juice of bishkatali showed 69.23% efficacy at 20% concentration against Trichomonas gallinae, in
vitro
condition. In vivo condition, bishkatali showed 66.66% efficacy at 20% concentration. This result could not be compared
due to paucity of relevant literature. Bishkatali leaves contain flavonid and quercetin (Ghani, 2003) which have antiprotozoal
activity (Khalid et al., 1986). In vitro condition, Fresh juice of guava showed 42.85% efficacy at 20% concentration against
Trichomonas gallinae. On the other hand, in in vivo condition guava showed 41.66% efficacy at 20% concentration. This
result could not be compared and contrasted due to lack of relevant literature. But guava has several medicinal uses. Guava
leaves contain tannin (Ghani, 2003). Tannin has antiprotozoal activity (Daniel, 2005). Besides this, guava leaves contain
vitamin C, which also act as antioxidant (Kayser et al, 2002).
Commercial drugs
During this study, five commercially available drugs, namely, metronidazole, tinidazole, ornidazole, secnidazole, nitazoxanide were used. In vitro and in vivo condition, different commercial drugs showed different efficacy in different doses. Metronidazole, ornidazole & secnidazole (30mg/kg) showed the highest efficacy (100%) whereas nitazoxanide (20mg/kg) was the least effective (84.61%) in in vitro condition. In vivo condition, metronidazole, ornidazole and secnidazole (30mg/kg) also showed the highest efficacy (100%) whereas tinidazole and nitazoxanide (20mg/kg) were the least effective (83.33%). Table 3: Efficacy of commercial drugs against Trichomonas gallinae in in vitro condition χ2 value
Table 4: Efficacy of commercial drugs against Trichomonas gallinae in in vivo condition χ2 value
These findings are supported by the findings of some other scientists in different parts of the world. Munoz et al. (1998)
showed a greater potency of ornidazole than the nitroimidazole derivatives against Trichomonas gallinae in in vitro condition.
Franssen and Lumeji (1992) also proved several nitroimidazoles including metronidazole, dimetridazole, ronidazole and
carnidazole, as effective drugs against Trichomonas gallinae in in vitro condition. Luo et al. (2006) observed that
dimetridazole and tinidazole were able to inhibit and control Trichomonas gallinae in in vitro condition. In Turkey, Aydn et
al
. (2000) described 100% and 87.5% efficacy rates for metronidazole and carnidazole, respectively against Trichomonas
gallinae
in naturally infected pigeons.
Cost effectiveness and highest efficacy for commercial drugs in Bangladesh aspect
Different drugs had different market prices. The prices were metronidazole 400mg/tab MRP=1tk and highest efficacy 100%, tinidazole 500mg/tab MRP=4tk and highest efficacy 94.11%, ornidazole 500mg/tab MRP=7tk and highest efficacy 100%, Int. J. BioRes. 2(11): 29-33 November, 2010 Biswas et al. secnidazole 1g/tab MRP=16tk and highest efficacy 100%, nitazoxanide 500mg/tab MRP=10tk and highest efficacy 92.30%. Metronidazole showed highest efficacy and cost effective than other commercial drugs. Table 5: Cost effectiveness and highest efficacy for commercial drugs in Bangladesh aspect %= percent; MRP= maximum retail price; tk= taka CONCLUSION
It was proved that commercial drugs were comparatively more effective than used herbal plants. The present study suggests that metronidazole is the most effective drugs tested against pigeon trichomoniasis and also cost effective. ACKNOWLEDGEMENTS
The authors thankfully acknowledge the financial assistance of the Bangladesh Agricultural University Research System (BAURES) to carry out this research. REFERENCES
Aydn, L., Coskum, S. and E. Gulegen. 2000. Efficacy of carnidazole (SpatrixReg.) and metronidazole (FlagylReg.) against Trichomonas gallinae in naturally infected pigeons. Acta Parasitologica Turcica 24 (1):65-66. Baker, J. R. 1986. Trichomoniasis, a major cause of vomiting in budgerigars. Veterinary Record 18:447–449. Begum, N., Mamun, M. A. A., Rahman, S. A. and A. S. M. Bari. 2008. Epidemiology and pathology of Trichomonas gallinae in the common pigeon (Columba livia). Journal of the Bangladesh Agricultural University 6(2): 301-306. Catelli, E., Poglayen, G., Terregino, C., Orlando, C., Tonulli, A., Issa-Gadale, O., Roda, R. and A. Agnoletti. 1999. Survey of endoparasits of digestive tract of Columba livia (Gmelin, 1787) in Florence, Selezino Veterinaria 2: 75-85. Cole, R. A. 1999. Trichomoniasis. In: Friend M, Franson JC (eds), Field manual of wildlife diseases: general field procedures and diseases of birds. USDI, Geological Survey-BRD, Washington D C, USA, pp 201–206. Cooper, J. E. and S. J. Petty. 1988. Trichiomoniasis in free-living goshawks (Accipiter gntilis gentilis) from Great Britain. Journal of wildlife disease 24: 80-87. Daniel, M. 2005. Medicinal plants: chemistry and properties. Science Publishers, Enfield, New Hampshire, USA, pp. 225. Franssen, F. F. J. and J. T. Lumeij. 1992. In vitro nitroimidazole resistance of Trichomonas gallinae and successful therapy with an increased dosage of ronidazole in racing pigeons (Columba livia domestica). Journal of Veterinary Pharmacology and Therapeutics 15: 409-415. Ghani, A. 2003. Medicinal Plants of Bangladesh with Chemical Consituents and Uses. 2nd edn, Asiatic Society of Bangladesh, Ho fle, U., Blanco, J. M., Palma, L. and P. Melo. 2000. Trichomoniasis in Bonelli’s eagle (Hieraaetus fasciatus) nestlings in south-west Portugal. In: Lumeij J T, Remple J D, Redig P T, Lierz M and Cooper J E (eds), Raptor biomedicine III. Zoological Education Network, Lake Worth, Florida, pp 45–52. Kayser, O. F., Kiderlen, A. and S. L. Croft. 2002. Studies in Natural Product Research. Atta-Ur-Rahaman (editor), Freie University Press, Berlin, pp. 198. Khalid, S. A., Farouk, A., Geary, T. G. and J. B. Jensen. 1986. Potential anti-malarial candidate from African plants. An in vitro approach using Plasmodium falciparum. Journal of Etbnopharmacology 14: 45-51. Luo, F., Weng, Y., Zhang, J., Zhu, X. and G. Li. 2006. Efficacy of common antitrichomonal drugs against Trichomonas gallinae from pigeon in vitro. Veterinary Science in China 36(12): 980-982. Mesa, C. P., Stabler, R. M. and M. Berthrong. 1961. Histopathological changes in the domestic pigeon infected with Trichomonas gallinae (Jones’ Barn Strain) Avian Diseases 5:48–60. Mostafa, G. M. 1989. Methods of Statistics, 4th edition, Karim Press and Publication, Dhaka, pp. 296-299. Munoz, E., Castella, J. and J. F. Gutierrez. 1998. In vivo and in vitro sensitivity of Trichomonas gallinae to some nitroimidazole drugs. Veterinary Parasitology 78(8):239-246. Narcisi, E. M., Sevoian, M. and B. M. Honigberg. 1991. Pathologic changes in pigeons infected with a virulent Trichomonas gallinae strain (Eiberg). Avian Diseases 35:55–61. Int. J. BioRes. 2(11): 29-33 November, 2010 Biswas et al. Real, J., Manosa, S. and E. Munoz. 2000. Trichomoniasis in a Bonelli’s eagle population in Spain. Journal of Wildlife Richardson, U. F. and S. B. Kendall. 1963. Veterinary Protozology, 3rd edition, Oliver and Boyd, Edinburgh and London. Soulsby, E. J. L. 982. elminths, Arthropods and Protozoa of Domesticated Animals, 7th ed. Bailiere, Tindall and Cassel Ltd. Stabler, R. M. 1954. Trichomonas gallinae: a review. Experimental Parasitology 3:368–402. Toro, H., Saucedo, C., Borie, C., Gough, R. E. and H. Alcaino. 1999. Health status of free living pigeons in the city of Santiago. Avian Pathology 28(6): 619-623.

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