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46(4) 295~310, 2017 295 綜論報告常見之球蟲藥及其使用規範 (1) 辛坤鎰 (2)(3) 林美峰 摘要 : 1990 ( ) McEvoy, 2002 (1) Integrated Open Systems Unit, Okinawa Institute of Science and Technology Graduate University. 1919-1 Tancha, Onna-son, Okinawa, Japan 904-0495. (2) 10672 3 155 50 (3) E-mail: mflin@ntu.edu.tw

296 緒 言 Waggoner et al., 1994 polyether ionophores synthetic coccidiostatschapman, 2007 monensin narasin lasalocid salinomycin maduramicin lasalocid analogues Lasalocid A B C D E Lasalocid A B C D E 10% halofuginonerobenidine diclazuril nicarbazin Yakkundi et al., 2002; McEvoy et al., 2003 tiamulin Umemura et al., 1985; Sakar et al., 1991; Croubels et al., 2001; Islam et al., 2009 1990 Committee for Medicinal Products for Veterinary Use; EMA/CVMP/EWP/706095/2015 球蟲病 (Coccidiosis) 之簡介 Eimeria spp. Isospora spp. Rypula et al., 2012Shirley and Lillehoj, 2012Mitchell et al., 2012Chartier and Paraud, 2012 Akpo et al., 2012

297 asexual cycle oocystsporozoitestrophozoitesschizonts merozoites male/female gametes sexual cycleoocyst sporulation Peek and Landman, 2011 球蟲藥之化學屬性 amprolium 啉 / diclazuril 50 streptomycetaceae 1967 Laidlomycin 呋 tetrahydrofuran rings spiroketal pharmacophore 1 Na + K + complexmichał Antoszczak and Huczynski, 2015 喹啉 quinolones 吡 pyridones alkaloidsguanidines triazine B 1 thiamine 2 喹啉 1960 quinoline nucleusbuquinolate Raines, 1968imine C=N Toltrazurilsix-membered heterocyclic ringsymmetric 1,3,5 asymmetric 1,2,4

298 1 呋 tetrahydrofuran rings spiroketal Figure 1 Chemical structures of polyether ionophores, featured in the tetrahydrofuran rings and spiroketal groups. 2 quinolones 喹啉 pyridones 吡 alkaloids guanidines triazine thiamine B 1 Figure 2 Chemical structures of synthetic coccidiostats. The major functional groups include quinolones, pyridines, alkaloids, guanidines, triazine derivative and thiamine analogue.

299 B1 3 B1 噻 thiamine pyrophosphotransferase 4 B1 B1 B1 Hamamoto et al., 2000 hydroxyethyl functionality B1 phosphorylationpyrophosphate B1 B1 B1 Bauchop and King, 1968; Patton et al., 1984> 0.07% B1 B1 yolk membrane Polin et al., 1963 3 Amprolium Thiamine B 1 Figure 3 Structural comparison of Amprolium (A) and Thiamine (B).

300 4 Thiamin-Pyrophosphokinase PDB ID: 1IG0Thiamin pyrophosphokinase amprolium docking simulation thiamin Figure 4 Structure of Thiamin-Pyrophosphokinase complex (PDB ID: 1IG0). Thiamin is shown as cyan compound, bound to the active site of pyrophosphokinase. Amprolium is shown as red. The computational docking simulation demonstrated that both thiamin and amprolium molecules competitively target on the same binding site. 球蟲藥之作用機制 一 聚醚類離子型球蟲藥 : Na + K + Ca 2+ Na + substrate oxidationatp hydrolysis Na + Ca 2+ Kart and Bilgili, 2008 ion carriers channel formers hydrophobic lipid bilayer 4 hydrogen bond donor 8 hydrogen bond acceptorfunctional group 2D lasalocid A, B, C, D E lasalocid-strontium complex 3D 5

301 6 Akkurt et al., 2007 Na + /K + gradient R1 R2 R3 R4 Lasalocid A Methyl Methyl Methyl Methyl Lasalocid B Ethyl Methyl Methyl Methyl Lasalocid C Methyl Ethyl Methyl Methyl Lasalocid D Methyl Methyl Ethyl Methyl Lasalocid E Methyl Methyl Methyl Ethyl 5 Lasalocid 2D (Akkurt et al., 2007) Figure 5 Chemical structure of Lasalocid and its analogues (Akkurt et al., 2007). 6 Lasalocid-strontium 3D lasalocid OH Figure 6 Structure of Lasalocid-strontium complex. Two lasalocid molecules interact with strontium (green) through a number of OH groups (red).

302 stereochemistry binding selectivity cavity Na +, K + Ca 2+, Mg 2+ Michał Antoszczak and Huczynski, 2015 peptidoglycan layer cytoplasmic membrane Gram-negative bacteria lipopolysaccharide layer outer membrane periplasmic space Kart and Bilgili, 2008 gramicidin hydrophilic channel Bergen and Bates, 1984 二 化學合成型球蟲藥 second-generation schizonts B 1 B 1 James, 1980 ethopabate 25:1.6 Clopidol decoquinate 喹 喹 nequinate Fry and Williams, 1984 folic acidparaaminobenzoic acid; PABA nicarbazin 1950 NAD + respiratory chain - C succinate-cytochrome C reductasedihydrofolate reductase Harder and Haberkorn, 1989 球蟲之防治計畫及成效 cross resistance :

303 一 單一投藥計畫 (single drug program) 喹啉 二 替換投藥計畫 (shuttle program) starter feed grower feed 三 循環投藥計畫 (rotation program) successive flocks clean-out Chapman et al., 2010 7 Chapman and Jeffers, 2014

304 7 rotation programchapman and Jeffers, 2014 6 1+2 5+6 shuttle program 3+4 drug-sensitive strains Figure 7 Proposal of a rotation program for a yearly broiler production cycle, involving six successive flocks. The program is composed of the application of chemotherapy and vaccine together with litter removal (clean out) after the second flock. The use of chemotherapy in the 1+2 and 5+6 flocks (blue and red) can include synthetic drugs, ionophores or the combination of both two classes. The application of vaccine can repopulate the drug-sensitive strains of coccidia with the hope to replace the drug-resistant strains in the broiler house. botanical extracts Arczewska- Włosek and Świ tkiewicz, 2012essential oil of oregano polyphenol carvacrol thymol antimicrobial

305 antioxidant activitiesbozkurt et al., 2013 ATP Ultee et al., 2002 Mohiti- Asli and Ghanaatparast-Rashti, 2015 Arczewska-Włosek and Świ tkiewicz, 2013; Bozkurt et al., 2016; Arczewska-Włosek et al., 2017 CocciVac 1952 subunit vaccines live vaccines DNA Peek and Landman, 2011 attenuatednon-attenuated sporulated oocysts / hatchery spray 14 Shirley et al., 2005; Arczewska-Włosek et al., 2017 Lehman et al., 2009 secondary enteritis CP 2%Arczewska- Włosek and Świ tkiewicz, 2014 球蟲藥之殘留及使用規範 1990 Kennedy et al., 1998non-compliant residues Huet et al., 2005 cross-contamination Glenn Kennedy et al., 1998; O Keeffe et al., 2007 Kennedy et al., 2000European Food Safety Authority EFSAmaximum residue levels

306 MRLs for target species by Commission Regulation No. 124/ 2009 Maximum Limits MLs for non-target species 1 1 Table 1 Tolerance limits of coccidiostats regulated by EU, US, FAO/WHO, Australia and Canada Substance Amprolium Hydrochloride a Tolerance limit (ug/kg) Regulations Muscle Skin and fat in natural proportions Liver Kidney Egg EU 200 200 200 400 1000 US 500-1000 1000 4000 Canada 500-1000 1000 7000 Australia 500-1000 1000 4000 EU 60 300 300 150 150 US - 1200 400 - - Lasalocid b Canada 100 400 400 400 - FAO/WHO 400 600 1200 600 - EU 30 150 150 100 - Maduramicin c US 240 480 720 - - Canada 100 400 500 1000 - EU 4000 4000 15000 6000 - US 4000 4000 4000 4000 - Nicarbazin d Canada 4000 4000 15000 800 - FAO/WHO 200 200 200 200 - EU 200 1300 800 350 - Robenidine 200 400 400 200 - Hydrochloride e Canada 100 200 100 100 - a: EU: Council Regulation (EEC) No. 2377/90. US: NADA Number: 013-149. Canada: Marketing Authorization for Maximum Residue Limits for Veterinary Drugs in Foods (August 2, 2017). Australia: 8 ppm in egg yolks. Codex; CX/RVDF 00/13. b: EU: EMA/CVMP/769137/2014. US: NADA Number: 096-298. Canada: same as above. FAO/WHO: Expert Committee on Food Additives. World Health Organization technical report series, 1 (2014). c: EU: Regulation (EU) No 388/2011. US: FDA 21CFR556.375. Revised as of April 1, 2015. Canada: same as above. d: EU: EFSA Journal 2010; 8(3):1551. US: FDA ANADA 200-478. Canada: same as above. FAO/WHO: Technical Report Series, No. 888, 1999. e: EU: The EFSA Journal 798, 1-15 (2008). Chickens for fattening and turkeys, respectively. Canada: same as above.

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J. 310 Chin. Soc. Anim. Sci. 46(4): 295~310, 2017 Review Coccidiostats and the regulations of their use Kun-Yi Hsin (1) and Mei-Fong Lin (2)(3) ABSTRACT Coccidiostats are broad-spectrum antibiotics used as anticoccidial agent and growth promotant in practice, commonly administered in animal feed for intensive production at recommended doses. The application of coccidiostats and their residues in food products are of increasing concern because of food safety issue. We summarize the chemical properties of several commonly used coccidiostats together with the mechanisms of action, anticoccidial programs and the relevant regulations of use. (Key Words: Coccidiosis, Polyether ionophores, Synthetic coccidiostats) (1) Integrated Open Systems Unit, Okinawa Institute of Science and Technology Graduate University. 1919-1 Tancha, Onna-son, Okinawa, Japan 904-0495. (2) Department of Animal Science and Technology, National Taiwan University, No.50, LN. 155, Sec. 3 Keelung Rd., 10672 Taipei, Taiwan. (3) Correspondence author, E-mail: mflin@ntu.edu.tw