Anti-protozoan study of a medicinal herb, Bidens pilosa

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1 2017 JITMM Anti-protozoan study of a medicinal herb, Bidens pilosa Meng-Ting Yang, Tien-Fen Kuo, Yueh-Chen Wu, Cicero L.T. Chang and Wen-Chin Yang Taiwan International Graduate Program Molecular and Biological Agricultural Sciences Academia Sinica, Taipei, Taiwan

Outline 2 Introduction Tropical diseases and medicine Coccidiosis Current approaches to coccidiosis control Results Anti-coccidial properties of BP Anti-coccidial MOA of BP Conclusions

Introduction 3

Tropical diseases and medicine 4 Tropical diseases are infectious diseases that are prevalent in or unique to tropical and subtropical regions. They kill tens of millions of people every year and become one of the greatest challenges of the 21st century. Tropical medicine is important to global health and food safety. Topical diseases Bacteria Virus Protozoan Helminth Food animal production http://www.tropicalmedicine.ox.ac.uk/research; http://www.jitmm.com/client/displaycontent.php?cpid=1

Coccidiosis in chickens 5 Avian coccidiosis, a protozoan disease, is one of the most serious infectious diseases in poultry (Yang, W.C. (2016) ecam 2657981). Annual production of chickens is estimated to be 50 billion with a market worth of 60 billion USD. Economic loss caused by coccidiosis is estimated to cost over 800 million USD annually (Intl J Poultry Sci 2004, 3(11): 715-718). Coccidia (Eimeria species) 34% protein food for human Most of poultry industry is in topical country Noninfected chickens Infected chickens Looks Stools Gut lesions http://www.betabunny.com/predators/predators_bonus.htm; http://darwinian-medicine.com/protein-fat-and-carbohydrate-how-much-of-each-should-you-eat/

The life cycle of Eimeria species 6 Coccidiosis is caused by 9 Eimeria species (Yang, W.C. (2016) ecam 2657981). Evidence-Based Complementary and Alternative Medicine, Volume 2016, Article ID 2657981, 19 pages; BioMed Research International Volume 2015, Article ID 430610, 11 pages; http://www.thepoultrysite.com/articles/3615/necrotic-enteritis-coccidiosis-multifactorial-role-of-butyrate-on-intestinal-health/

Current approaches to coccidiosis control 7 Usage Drugs Vaccines Phytoagents Prevention Treatment Prevention Prevention Treatment Drug resistance + - ± Drug residue + - - Mode of action + +? Preventive use of antibiotic chemicals in poultry will be banned by 2021 in Europe and 2017 in USA. Thus, edible plants are emerging as attractive way to control coccidiosis. H. D. Chapman, Department of Poultry Science, University of Arkansas, Vet Parasitol (2011)181(2-4):97-105, Parasitol Int (2010) 59(4):506-11, Poult Sci (1997) 76(8):1156-63, Vet Parasitol (2008) 153(3-4):214-9, Vet Parasitol (2011) 182(2-4):121-6, Vet Parasitol (2012)186(3-4):170-7, Vet Parasitol (2012) 185(2-4):158-63, Parasitol Res (2003) 91(1):74-8, Vet Parasitol (2001) 96(4):257-63.

Herbal medicine-bidens pilosa Clinical experience for thousands years. Plant or plant-derived medicines are used in different categories of diseases. 8 Bidens pilosa (BP): Edible, palatable and easy-to-grow plant. Tropical or subtropical areas. It is listed as food by FAO of UN and MOHW of Taiwan. It has been reported to treat over 40 categories of diseases such as protozoan infection, bacterial infection, gut disorders, immune disorders, etc. The anti-coccidial property of BP is still elusive. Food and Agriculture Organization of the United Nations (FAO) The Ministry of Health and Welfare (MOHW) Evidence-Based Complementary and Alternative Medicine, Volume 2016, Article ID 2657981, 19 pages

Results 9

Anti-coccidial efficacy of BP in chickens. 10 0 Days Post-hatch 1 14 18 21 PBS Et Et+Mad Et+BP Et challenge OPG PBS, mad and BP feeding Survival rate (D 14 to 21) Body weight (D 14 to 21) Gut pathology (D 21) Mad:maduramycin BP:Bidens pilosa OPG:oocysts per gram of feces Survival Rate (%) 100 90 80 70 60 0 PBS (n=13) Et(n=15) Et+ Mad(n=15) Et+ 5%BP (n=12) Et+ 1% BP (n=13) Et+ 0.1% BP (n=9) 1 2 3 4 5 6 7 Days of post-infection P = 0.01 P = 0.03

BP increases body weight of chickens. 11 1. The chickens were given standard diet (Groups 1 and 2) and standard diet supplemented with 0.5% B. pilosa product (Groups 3 and 4) from days 1 to 21. On day 14, chickens in Groups 2 and 4 were orally inoculated with E. tenella at the dose of 1 10 4 sporulated oocysts per chicken. 2. Body weight gain (%) was calculated based on the formula: 100% (body weight on day 21 minus body weight on day 14)/body weight on day 14. 3. FCR stands for feed conversion ratio and it was obtained by normalization of feed intake to body weight gain. PLOS ONE DOI:10.1371/journal.pone.0146141 January 14, 2016

BP decreases gut pathology of chickens. 12

Field trial of BP on coccidiosis in chickens. 13

14 BP suppresses the invasion of E. tenella sporozoites into gut cells.

Effect of BP on the change of gut microbiota in chickens. 7 beneficial bacteria 15 harmful bacteria 15 Bilophila Lactobacillus Alistipes Rikenella Ruminococcus2 Megamonas Bacteroides Acinetobacter Clostridium IV Anaerostipes Anaeroplasma Enterococcus Campylobacter Flavonifractor Escherichia/Shigella Oscillibacter Pseodoflavonifractor Odoribacter Phascolarctobacterium Anaerotruncus Butyricicoccus Clostridium XIVb

A schematic model of BP in coccidiosis of chickens. 16 Life cycle of coccidia Gut microbiota Beneficial BP Harmful Anti-coccidial functions Other functions Mechanisms Red word: up/ Blue word: down

Conclusions 17 BP can control coccidiosis via multiple mechanisms, including interference with of Eimeria life cycle and modulation of gut bacteria. BP can serves as a novel remedy for coccidiosis in chickens.

18 Acknowledgement Dr. Wen-Chen Yang and lab crew Dr. Cicero L.T. Chang

19

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Thank you 21