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Anticoccidial Activity of Traditional Chinese Herbal Dichroa Febrifuga Lour. Extract Against Eimeria Tenella Infection in Chickens

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Anticoccidial Activity of Traditional Chinese Herbal Dichroa Febrifuga Lour. Extract Against Eimeria Tenella Infection in Chickens Parasitol Res (2012) 111:2229–2233 DOI 10.1007/s00436-012-3071-y ORIGINAL PAPER Anticoccidial activity of traditional Chinese herbal Dichroa febrifuga Lour. extract against Eimeria tenella infection in chickens De-Fu Zhang & Bing-Bing Sun & Ying-Ying Yue & Qian-Jin Zhou & Ai-Fang Du Received: 27 April 2012 /Accepted: 30 July 2012 /Published online: 17 August 2012 # Springer-Verlag 2012 Abstract The study was conducted on broiler birds to evalu- use of anticoccidial drugs (Hao et al. 2007). The domestic ate the anticoccidial efficacy of an extract of Chinese traditional poultry industry of People's Republic of China primarily relies herb Dichroa febrifuga Lour. One hundred broiler birds were on medical prophylaxis. But the emergence of problems re- assigned to five equal groups. All birds in groups 1–4were lated to drug resistance and drug residues of antibiotics in the orally infected with 1.5×104 Eimeira tenella sporulated chicken meat has stimulated us to seek safer and more effica- oocysts and birds in groups 1, 2 and 3 were medicated with cious alternative control strategies (Lai et al. 2011). 20, 40 mg extract/kg feed and 2 mg diclazuril/kg feed, respec- Chinese traditional herbal medicines have been utilized for tively. The bloody diarrhea, oocyst counts, intestinal lesion human and animal health for millenniums. Currently, phyto- scores, and the body weight were recorded to evaluate the therapies are investigated as alternative methods for control- anticoccidial efficacy. The results showed that D. febrifuga ling coccidian infections. A number of herbal extracts have extract was effective against Eimeria infection; especially been proven to be efficient to control coccidiosis. Metwaly et 20 mg D. febrifuga extract/kg feed can significantly increase al. (2012) reported the effect of Khodary date fruit extract body weight gains and reduce bloody diarrhea, lesion score, from Phoenix dactylifera on the coccidiosis caused by Eime- and oocyst excretion in comparison to infected-unmedicated ria papillata. The date fruit extract can decrease the fecal control group. oocyst output and reduce the parasites in the intestinal villi and associated histopathological lesions. It was evaluated that curcumin (diferuloylmethane), which is extracted from the Introduction perennial herb turmeric (Curcuma longa) can inhibit Eimeria tenella sporozoites viability and infectivity in vitro (Khalafalla Coccidiosis may cause severe intestinal diseases. Coccidiosis et al. 2011). del Cacho et al. (2010)showedthatartemisinin is distributed worldwide in poultry and wild birds and is of extracted from the natural herb Artemisia annua can alter the major economic impact in poultry production due to mortality oocyst wall formation, reduce the sporulation rate, and inhibit and morbidity. Epidemiologic studies have shown that the expression of sarcoplasmic–endoplasmic reticulum calcium mortality of fowl can range from 5 to 70 % (Du and Hu ATPase in macrogamete stage. De Pablos et al. (2010)studied 2004; Michels et al. 2011). Economic losses caused by avian the anticoccidial efficacy of maslinic acid. The results showed coccidiosis, including decreased productivity, usage of cocci- the anticoccidial index (ACI) for maslinic acid was 210.27 diostat, and vaccines, are estimated to amount to $3 billion compared 173.09 for control treatment. Maslinic acid can annually (Dalloul and Lillehoj 2006; Lai et al. 2011; Williams decrease lesions and oocyst counts and increase the body 1999; Zhang et al. 2012b; Michels et al. 2011). In China alone, weight. Naidoo et al. (2008) demonstrated that three plant the estimated expenditure is about $30–60 million due to the extracts, Tulbaghia violacea, Vitis vinifera, and Artemisia afra, can improve the feed conversion ratios of Eimeria- : : : : infected chickens in comparison to the untreated control group D.-F. Zhang B.-B. Sun Y.-Y. Yue Q.-J. Zhou A.-F. Du (*) and T. violacea significantly decreases oocyst excretion. Institute of Preventive Veterinary Medicine, Michels et al. (2011) also proved that infected chickens when College of Animal Sciences, Zhejiang University, Hangzhou 310058, People’s Republic of China treated with coumestans from Eclipta alba displayed higher e-mail: [email protected] weight gains, improved feed efficiency and significantly 2230 Parasitol Res (2012) 111:2229–2233 decreased fecal oocysts and macroscopic lesions. Youn and 500 ml chloroform at room temperature for 1 day. The Noh (2001) screened 15 different Asian herbal extracts for mixtures were heated with reflux of chloroform for 2 h. activity against E. tenella. Extract from Sophora flavescens After filtration, the residues were suspended in 500 ml chlo- Aiton was the most effective preparation as indicated by roform and extracted. The combined solvent was evaporated survival rates, diarrhea scores and lesion scores, body by heating. The extracted residue was grounded and stored weight, and oocyst excretion. Many other plant extracts, in sealable plastic bag at 4 °C until use. such as tannins, xanthohumol, flaxseed oil, and betaine, have been screened to be effective in controlling Eimeria Experimental design infection in vitro or in vivo (Molan et al. 2009; Allen et al. 1996, 1997; Allen 2007; Augustine et al. 1997;Augustine A total of one hundred 12-day-old birds were weighed and and Danforth 1999). randomly distributed into five groups of 20 birds. Birds in Halofuginone hydrobromide is a synthesized analog of groups 1 and 2 were provided with 20 or 40 mg extract in febrifugine, a constituent of the leaf and root of Chinese 1 kg feed from day 12 to day 21. Group 3 was treated with medicinal herb Dichroa febrifuga Lour. (Chinese name 2 mg diclazuril/kg feed from day 14 to day 21, while group Cháng Shān), which is an anticoccidial drug approved by 4 and group 5 served as untreated controls throughout the the European Union and the US Food and Drug Adminis- experiment. On day 14, birds in groups 1, 2, 3, and 4 were tration Agency (Zhang et al. 2012a). Therefore we have orally infected with 1.5×104 sporulated E. tenella oocysts. evaluated the anticoccidial effects of a chemical extract of The clinical signs were observed and recorded each day this herb as supplement in the broiler feed. until the end of the experiment. Evaluation of anticoccidial effect Materials and methods ACI½ ACI ¼ ðrelative ratio of body weight gains þ survival Birds and parasites rateÞðÞoocyst value þ median lesion scores was calculat- ed to evaluate the anticoccidial efficacy (Johnson and Reid One-day-old male broilers were bought from a local hatchery 1970;Maetal.2011). in Hangzhou, People's Republic of China. The birds were On the seventh day post-infection, the survival rate (SR) reared in clean wire cages in hygienically controlled of each group was calculated and birds were weighed to coccidian-free environment and provided with coccidiostat- determine the percentage increase in body weight gains free feed and water without any antibiotics. All procedures (piBWG) and relative ratio of body weight gains (rBWG). related to the animals and their care met the international SR was investigated as the number of surviving chickens/the guiding principles for biomedical research involving animals, number of initial chickens. The piBWG was calculated as issued by the Council for the International Organizations of as piBWG ¼ ðbody weight after infectionÀ body weight Medical Sciences (1985) and the World Association for before infectionÞ = ðÞÂbody weight before infection 100%. the Advancement of Veterinary Parasitology guidelines The rBWG was calculated for each group according to for evaluating the efficacy of anticoccidial drugs in chick- the following equation: rBWG ¼ ðÞpiBWG in each group = ens and turkeys as previously mentioned (Holdsworth et al. ðpiBWG in group of noninfected-unmedicated controlsÞ 2004) along with the Guidelines for Keeping Experimental 100%. Animals issued by the government of People's Republic of After being weighed, all surviving birds were subse- China. quently euthanized. The overall general appearances of ce- The sporulated oocysts of E. tenella Shanghai strain were cal lesions were recorded by two skilled persons who were kindly gifted by Dr. Bing Huang, Shanghai Veterinary Re- not informed on the group allocation of the respective birds. search Institute, China Academy of Agricultural Sciences. Lesion scores were assessed according to the method of The oocysts were stored in 2.5 % potassium dichromate Johnson and Reid (1970). Generally, cecal lesions were reagent at 4 °C and propagated in 2-week-old chickens ranked from 0 (normal) to 4 (dead or extremely severe) before use. based on the macroscopical appearance of the intestine. Oocysts in pooled caecum contents of each bird were Preparation of extract counted using a hemocytometer counting technique and the result was expressed as oocysts per gram (OPG) (9, According to the properties and previous described methods 11). Oocyst value ¼ ðÞOPG in each group =ðOPG in group of (Jiang et al. 2005; Murata et al. 1998), a modified chloro- infected-unmedicated controlsÞ100. Protection rate (in form reflux method was used to extract febrifugine. Briefly, percent) was calculated asð OPG from infected-unmedicated 100 g dry powder of D. febrifuga root was macerated in control birds À OPG from medicated birdsÞ = ðOPG from Parasitol Res (2012) 111:2229–2233 2231 infected-unmedicated control birdsÞ100% (Du and Wang 2005; Rose and Mockett 1983). Diarrhea scores were recorded between day 4 and day 7 post-infection according to Du and Hu (2004). Briefly, the percentage of samples containing visible blood was estimat- ed for each group and scored as follows: 0, no hemorrhagic samples; 1, 25 % of the samples are hemorrhagic; 2, 50 % of the samples are hemorrhagic; 3, 75 % of the samples are hemorrhagic; and 4, all the samples are haemorrhagic. Statistical analyses The data were analyzed using the software SPSS 16.0 for Windows (SPSS Inc., Chicago, IL). The parameters of body weight gains and OPG were analyzed by using one-way analysis of variance and mean values were compared using Fig.
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