臺灣博碩士論文加值系統

犬小焦蟲疾病（Babesia gibsoni）在臨床治療上仍面臨許多挑戰，在傳統及新的治療方法中似乎都沒有辦法完全根除，容易發現復發或是抗藥性的問題，而且治療方式無法利用單一藥物做到有效治療，往往都是複合式的雞尾酒療法才能達到治療效果。本實驗的研究目的為評估自然感染犬小焦蟲的犬隻分別使用clindamycin、doxycycline和metronidazole（簡稱CDM組）和diminazene加上CDM（簡稱DCDM）的治療效果比較。兩組分別為CDM組7隻，DCDM組9隻，由於DCDM組中有兩隻病犬進行輸血治療，故未列入血液學變化的統計分析，兩組在療程結束後，停藥兩周採血進行PCR檢測判定有無感染。CDM治癒率100%（7/7），DCDM為89%（8/9其中一隻死亡）。在血液學變化部分進行two-way ANOVA分析，結果為兩組之間並未發現對HCT、PLT的結果影響具有顯著差異（分別為p=0.09; p=0.44），但兩組之間對於HGB是影響具有顯著差異（p=0.016），但兩組別中HCT、HBG、PLT皆有顯著增加的趨勢，另外進行兩組的無母數分析發現第七天的HCT、HGB、PLT，兩組之間有顯著性差異（分別為p=0.013; p=0.019; p=0.037），表示DCDM組的HCT、HGB、PLT恢復狀況較CDM組治療後第七天有明顯回升。ALT，ALKP兩組分別有4/7、2/8在使用口服藥後出現上升的現象，而且兩組各有一隻犬隻的GGT上升超過正常值許多，但TBIL並無異常。最終結論是，兩組的治療結果沒有顯著性差異，而且效果良好，但是DCDM組於治療後第七天的恢復結果都比CDM組來的好，而治療過程中DCDM組發生一隻急性自體免疫性溶血而死亡，因為樣本數不夠多，無法推測是否與diminazene治療相關，因此CDM是犬小焦蟲症方便有效的治療選擇。

Canine babesiosis which is a protozoal, tickborne, hemolytic disease with worldwide distribution and global significance and specially caused by Babesia gibsoni is a big challenge and critical issue in the treatment until right now. It seems not to be complete eradicated by a traditional single drug or some new combination treatments such as azithromycin with atovaquone, and clindamycin, diminazene, with imidocarb, and enrofloxacin, doxycycline with metronidazole. After treatment, canine babesiosis is easily to relapse again or have drug-resistant problems which have been studied recently. The combination treatment protocol is more efficiency than the single drug administrated to canine babesiosis in most studies. It is the life-threaten and emergency disease which needs the appropriate treatment in dogs. The aim of this study is to compare the efficacy of a combination of clindamycin, doxycycline, and metronidazole (CDM) to that of diminazene and clindamycin-doxycycline-metronidazole (DCDM) for the treatment of natural B. gibsoni infections, 16 client-owned dogs were random collected in the study. Seven dogs were treated with CDM combination and 9 dogs were treated with DCDM. Seven of the 7 dogs in the CDM group and 8 of the 9 dogs in the DCDM one completely recovered, but one of the DCDM group was died in acute IMHA after received the diminazene injection. The statistical differences in the numeric variables (HCT, HGB, and PLT) between CDM and DCDM groups were noticed only at the 7th day (p=0.013; p=0.019; p=0.037, respectively) after assessed by Wilcoxon’s rank-sum test. The changes of the serum biochemisty after treatment were shown that the ALT, and ALKP levels were elevated higher than first day (4/7 and 2/8, respectively). There is one dog individual in each group with highly GGT after treatment, but the TBIL of all dogs were normal in the treatment period. The conclusion is that it is good efficacy and no significate difference in both groups but diminaneze is not available in many countries in order to CDM will be the choice of treatment for B. gibsoni. Although the CDM treatment protocol might elevate the serum liver enzyme levels, it is not appeared serious clinical signs with vomit or dropped appetite. After combine with ursodiol and silymarin treatment, the serum live enzyme levels return back to the normal range which may means CDM treatment should be used with caution in animal with liver dysfunction.

摘 要 i
Abstract ii
目 次 iv
表 次 vii
圖 次 viii
第一章 緒言 1
第二章 文獻探討 3
第一節 壁蝨簡介 3
一、 壁蝨分類、分布與媒介性疾病關係 3
二、 壁蝨媒介性疾病致病機制 4
三、 壁蝨媒介性疾病傳播時間 6
第二節 焦蟲簡介 8
一、 焦蟲分類 8
二、 焦蟲生活史 9
三、 焦蟲特殊結構與侵入血球機制 11
四、 感染小焦蟲（B. gibsoni）的臨床症狀與合併感染 15
第三節 治療焦蟲藥物作用機制 17
第四節 Diminazene 20
一、 藥物介紹 20
二、 藥理機制及藥物作用 21
第五節 Clindamycin 21
一、 藥物介紹 21
二、 藥理機制及藥物作用 22
第六節 Doxycycline 23
一、 藥物介紹 23
二、 藥理機制及藥物作用 24
第七節 Metronidazole 25
一、 藥物介紹 25
二、 藥理機制及藥物作用 25
第三章 材料與方法 27
第一節 實驗動物 27
第二節 實驗方法 29
一、 藥物選擇 29
二、 給藥方式 29
三、 實驗方法 30
四、 統計分析 31
第四章 實驗結果 33
第一節 正式實驗 33
一、 CDM與DCDM組間犬隻體重及年齡比較 33
二、 CDM與DCDM組間治療後血容比（HCT）變化 36
三、 CDM與DCDM組間治療後血紅素濃度（HGB）變化 38
四、 CDM與DCDM組間治療後血小板濃度（PLT）變化 40
五、 CDM與DCDM組間治療後寄生蟲血症（parasitemia）變化 42
六、 CDM與DCDM組間治療後血液生化值變化 43
七、 CDM與DCDM組間治癒率及自體免疫性溶血性貧血發生率 48
第五章 討論 49
參考文獻 54
附錄一 60
附錄二 61
附錄三 62
附錄四 68
附錄五 74

1. Wu T, Sun H, Wu Y, et al. Prevalence and risk factors of canine ticks and tick-borne diseases in Taipei, Taiwan. J Vet Clin Sci. 2009;2:75-78.
2. Matsuu A, Koshida Y, Kawahara M, et al. Efficacy of atovaquone against Babesia gibsoni in vivo and in vitro. Vet Parasitol. 2004;124:9-18.
3. Wulansari R, Wijaya A, Ano H, et al. Clindamycin in the Treatment of Babesia gibsoni Infections in Dogs. J Am Anim Hosp Assoc. 2003;39:558-562.
4. Suzuki K, Wakabayashi H, Takahashi M, et al. A Possible Treatment Strategy and Clinical Factors to Estimate the Treatment Response in Bebesia gibsoni Infection. J Vet Med Sci. 2007;69:563-568.
5. Birkenheuer A. Babesia gibsoni infections in dogs from North Carolina. J Am Anim Hosp Assoc. 1999;35:125-128.
6. Lin E, Chueh L, Lin C, et al. The therapeutic efficacy of two antibabesial strategies against Babesia gibsoni. Vet Parasitol. 2012;186:159-164.
7. Lin M, Huang H. Use of a doxycycline-enrofloxacin-metronidazole combination with/without diminazene diaceturate to treat naturally occurring canine babesiosis caused by Babesia gibsoni. Acta Vet Scand. 2010;52:27.
8. Matsuu A, Miyamoto K, Ikadai H, et al. Cloning of the Babesia gibsoni cytochrome b gene and isolation of three single nucleotide polymorphisms from parasites present after atovaquone treatment. Am J Tro Med Hy. 2006;74:593-597.
9. Homer MJ, Aguilar-Delfin I, Telford SR, et al. Babesiosis. Clin Microbiol Rev. 2000;13:451-469.
10. Boozer A, Macintire D. Canine babesiosis. Vet Clin North Am Small Anim Pract. 2003;33:885-904.
11. Wulansari R, Wijaya A, Ano H, et al. Lymphocyte Subsets and Specific IgG Antibody Levels in Clindamycin-Treated and Untreated Dogs Experimentally Infected with Babesia gibsoni. J Vet Med Sci. 2003;65:579-584.
12. Vial H, Gorenflot A. Chemotherapy against babesiosis. Vet Parasitol. 2006;138:147-160.
13. Birkenheuer A, Levy M, Breitschwerdt E. Efficacy of Combined Atovaquone and Azithromycin for Therapy of Chronic Babesia gibsoni (Asian Genotype) Infections in Dogs. J Vet Intern Med. 2004;18:494-498.
14. Shaw S, Day M, Birtles R, et al. Tick-borne infectious diseases of dogs. Trends Parasitol. 2001;17:74-80.
15. Willadsen P, Jongejan F. Immunology of the Tick–Host Interaction and the Control of Ticks and Tick-borne Diseases. Parasitol Today. 1999;15:258-262.
16. Ferreira B, Silva J. Successive tick infestations selectively promote a T-helper 2 cytokine profile in mice. Immunology. 1999;96:434-439.
17. Labuda M, Kozuch O, Zuffová E, et al. Tick-Borne Encephalitis Virus Transmission between Ticks Cofeeding on Specific Immune Natural Rodent Hosts. Virology. 1997;235:138-143.
18. Irwin PJ, Hutchinson GW. Clinical and pathological findings of Babesia infection in dogs. Aust Vet J. 1991;68:204-209.
19. Adachi K, Yoshimoto A, Hasegawa T, et al. Anti-Erythrocyte Membrane Antibodies Detected in Sera of Dogs Naturally Infected with Babesia gibsoni. J Vet Med Sci. 1992;54:1081-1084.
20. Morita T, Saeki H, Imai S, et al. Erythrocyte oxidation in artificial Babesia gibsoni infection. Vet Parasitol. 1996;63:1-7.
21. Alleman R. Understanding the Transmission of Tick-borne Pathogens. continue education; 2012; AAHA.
22. McQuiston J, Childs J, Chamberland M, et al. Transmission of tick-borne agents of disease by blood transfusion: a review of known and potential risks in the United States. Transfusion. 2000;40:274-284.
23. Shaw S. Understanding Transmission of Infection by Ticks and New Strategies for Control. Paper presented at: World Small Animal Veterinary Association World Congress Proceedings 2008; Dublin, Ireland
24. Levine N. Progress in taxonomy of the Apicomplexan protozoa. J protozool. 1988;35:518-520.
25. Levine N. Erhardorina n. g., Ascogregarina polynesiensis n. sp., Eimeria golemanskii n. sp., Isospora tamariscini n. sp., Gregarina kazumii n. nom., New Combinations and Emendations in the Names of Apicomplexan Protozoa. J protozool. 1985;32:359-363.
26. Uilenberg G. Babesia - A historical overview. Vet Parasitol. 2006;138:3-10.
27. Carret C, Walas F, Carcy B, et al. Babesia canis canis, Babesia canis vogeli, Babesia canis rossi: Differentiation of the Three Subspecies By A Restriction Fragment Length Polymorphism Analysis On Amplified Small Subunit Ribosomal Rna Genes. J Eukaryot Microbiol. 1999;46:298-301.
28. Kakoma I, Mehlhorn H. Babesia of domestic animals. In: Kreier JP, ed. Parasitic Protozoa. Vol 7. second ed. San Diego: Academic Press; 1993:141-216.
29. Valentin A, Rigomier D, Precigout E, et al. Lipid trafficking between high density lipoproteins and Babesia divergens-infected human erythrocytes. Biol cell. 1991;73:63-70.
30. Mackenstedt U, Gauer M, Mehlhorn H, et al. Sexual cycle of Babesia divergens confirmed by DNA measurements. Parasitol Res. 1990;76:199-206.
31. Birkenheuer A. Babesiosis. In: Greene, ed. Infectious diseases of the dog and cat. fourth ed. St. Louis, Missouri: Elsevier saunders; 2012:771-784.
32. Kemp L, Yamamoto M, Soldati-Favre D. Subversion of host cellular functions by the apicomplexan parasites. FEMS Microbiology Reviews. 2013;37:607-631.
33. Samuelson J. Why Metronidazole Is Active against both Bacteria and Parasites. Antimicrob Agents Chemother. 1999;43:1533-1541.
34. Frénal K, Polonais V, Marq J, et al. Functional Dissection of the Apicomplexan Glideosome Molecular Architecture. Cell Host Microbe. 2010;8:343-357.
35. Paredes-Santos T, de Souza W, Attias M. Dynamics and 3D organization of secretory organelles of Toxoplasma gondii. J Struct Biol. 2012;177:420-430.
36. Carruthers V, Sibley L. Mobilization of intracellular calcium stimulates microneme discharge in Toxoplasma gondii. Mol Microbiol. 1999;31:421-428.
37. Carruthers V, Tomley F. Receptor-ligand interaction and invasion: Microneme proteins in apicomplexans. Subcell biochem. 2008;47:33-45.
38. Friedrich N, Matthews S, Soldati-Favre D. Sialic acids: Key determinants for invasion by the Apicomplexa. Int J Parasitol. 2010;40:1145-1154.
39. Kessler H, Herm-Götz A, Hegge S, et al. Microneme protein 8 – a new essential invasion factor in Toxoplasma gondii. J Cell Sci. 2008;121:947-956.
40. Singh S, Alam M, Pal-Bhowmick I, et al. Distinct External Signals Trigger Sequential Release of Apical Organelles during Erythrocyte Invasion by Malaria Parasites. PLoS Pathog. 2010;6:1-14.
41. Shaw M, Tilney L, Musoke A. The entry of Theileria parva sporozoites into bovine lymphocytes: evidence for MHC class I involvement. J Cell Biol. 1991;113:87-101.
42. Gohil S, Kats L, Sturm A, et al. Recent insights into alteration of red blood cells by Babesia bovis: moovin’ forward. Trends Parasitol. 2010;26:591-599.
43. Sinai AP. Biogenesis of and activities at the Toxoplasma gondii parasitophorous vacuole membrane. Subcell Biochem. 2008;47:155-164.
44. Mital J, Meissner M, Soldati D, et al. Conditional expression of Toxoplasma gondii apical membrane antigen-1 (TgAMA1) demonstrates that TgAMA1 plays a critical role in host cell invasion. Mol biol cell. 2005;16:4341-4349.
45. Yokoyama N, Okamura M, Igarashi I. Erythrocyte invasion by Babesia parasites: Current advances in the elucidation of the molecular interactions between the protozoan ligands and host receptors in the invasion stage. Vet Parasitol. 2006;138:22-32.
46. Bork S, Yokoyama N, Ikehara Y, et al. Growth-inhibitory effect of heparin on Babesia parasites. Antimicrob Agents Chemother. 2004;48:236-241.
47. Suarez C, Laughery J, Bastos R, et al. A novel neutralization sensitive and subdominant RAP-1-related antigen (RRA) is expressed by Babesia bovis merozoites. Parasitology. 2011;138:809-818.
48. Meinkoth J, Kocan A, Loud S, et al. Clinical and hematologic effects of experimental infection of dogs with recently identified Babesia gibsoni-like isolates from Oklahoma. J Am Vet Med Assoc. 2002;220:185-189.
49. Collett M. Survey of canine babesiosis in South Africa. J S Afr Vet Assoc. 2000;71:180-186.
50. Van Heerden J, Reyers F, Stewart C. Treatment and thrombocyte levels in experimentally induced canine ehrlichiosis and canine babesiosis. Onderstepoort J Vet Res. 1983;50:267-270.
51. Kontos V, Kountinas A. Clinical observations in 15 spontaneous cases of canine babesiosis. Canine practice (Santa Barbara, Calif: 1990)(USA). 1997.
52. Adachi K, Ueno C, Makimura S. Immunosuppression in Dogs Naturally Infected with Babesia gibsoni. J Vet Med Sci. 1993;55:503-505.
53. Conrad P, Thomford J, Yamane I, et al. Hemolytic anemia caused by Babesia gibsoni infection in dogs. J Am Vet Med Assoc. 1991;199:601-605.
54. Slade D, Lees G, Berridge B, et al. Resolution of a proteinuric nephropathy associated with Babesia gibsoni infection in a dog. J Am Anim Hosp Assoc. 2011;47:e138-144.
55. Tasaki Y, Miura N, Iyori K, et al. Generalized alopecia with vasculitis-like changes in a dog with babesiosis. J Vet med Sci. 2013;75:1367-1369.
56. Reyers F, Leisewitz A, Lobetti R, et al. Canine babesiosis in South Africa: more than one disease. Does this serve as a model for falciparum malaria. Ann Trop Med Parasitol. 1998;92:503.
57. Welzl C, Leisewitz A, Jacobson L, et al. Systemic inflammatory response syndrome and multiple-organ damage/dysfunction in complicated canine babesiosis. J S Afr Vet Assoc. 2001;72:158-162.
58. Fichera M, Roos D. A plastid organelle as a drug target in apicomplexan parasites. Nature. 1997;390:407-409.
59. He C, Shaw M, Pletcher C, et al. A plastid segregation defect in the protozoan parasite Toxoplasma gondii. EMBO J. 2001;20:330-339.
60. Ralph S, D'Ombrain M, McFadden G. The apicoplast as an antimalarial drug target. Drug Resist Updat. 2001;4:145-151.
61. MacRae J, Maréchal E, Biot C, et al. The Apicoplast: A Key Target to Cure Malaria. Curr Pharm Des. 2012;18:3490-3504.
62. McDougald L, Roberson E. Antiprotozoan drugs. In: Booth NH, McDonald LE, ed. Veterinary pharmacology and therapeutics. sixth ed. Cedar Rapids (IA): Iowa State University Press; 1988:950-968.
63. Hatcher J, Greenberg P, Antique J, et al. Severe babesiosis in Long Island: review of 34 cases and their complications. Clin Infect Dis. 2001;32:1117-1125.
64. Wittner M, Lederman J, Tanowitz H, et al. Atovaquone in the treatment of Babesia microti infections in hamsters. Am J Trop Med Hyg. 1996;55:219-222.
65. Brasseur P, Lecoublet S, Kapel N, et al. In vitro evaluation of drug susceptibilities of Babesia divergens isolates. Antimicrob Agents Chemother. 1998;42:818-820.
66. Talukder M, Matsuu A, Hikasa Y. Evaluation of doxycycline sensitivity in vitro against Babesia gibsoni by Real Time-PCR. J Bangladesh Agril Univ. 2013;11:97-102.
67. Matsuu A, Yamasaki M, Xuan X, et al. In vitro evaluation of the growth inhibitory activities of 15 drugs against Babesia gibsoni (Aomori strain). Vet Parasitol. 2008;157:1-8.
68. Fowler J, Ruff M, Fernau R, et al. Babesia gibsoni: chemotherapy in dogs. Am J Vet Res. 1972;33:1109-1114.
69. Trepanier L. Applying Pharmacokinetics to Veterinary Clinical Practice. Vet Clin N Am-Small. 2013;43:1013-1026.
70. Milner R. The effect of diminazene aceturate on cholinesterase activity in dogs with canine babesiosis. J S Afr Vet Assoc. 2002;68:111-113.
71. Jacobson L, Lobetti R, Becker P, et al. Nitric oxide metabolites in naturally occurring. Vet Parasitol. 2001;104:27-41.
72. Kelly P, Koster L, Lobetti R. Canine babesiosis: a perspective on clinical complications, biomarkers, and treatment. Vet Med Res Rep. 2015:119.