臺灣博碩士論文加值系統

本實驗探討鴿子在注射環磷醯胺(CY)後，是否會因免疫系統受到抑制而提升對家禽流行性感冒病毒 (AIV)之感受性。以低病原性的台灣AIV分離株 A/chicken/Taiwan/1209/03 (H5N2)及A/chicken/Taiwan/3152/03 (H6N1)經眼鼻路徑接種，並利用巢式反轉錄聚合酶連鎖反應(nested RT-PCR)和雞胚胎蛋病毒分離等2種方式進行偵測。在H5N2及H6N1的感染試驗中，施打與無施打CY的鴿子，於21天的觀察期內皆無排毒及抗體陽轉之情形，並且無法在氣管、肺、胰、脾、腎及直腸等臟器檢測到病毒。H6N1試驗中的同居感染雞隻也為相同的陰性結果。因此推論鴿子在免疫抑制的情形下，不但仍對這2株AIV具有抗性，也不會媒介病毒的傳播使其它鴿子或雞隻感染。在本實驗中也利用H5N2 AIV進行肉雞的接種試驗，以探討病毒在組織臟器的殘留情形。結果顯示在感染後的5和7天，以nested RT-PCR可在氣管和直腸偵測到病毒，心肌及肌胃則在第5天，腦和骨髓的出現情形較少，胸肌、翅肌或腿肌皆為陰性。另外，在2004年10月到2005年4月間，對台灣地區994隻鴨隻、71隻鴿子以及80隻寵物鳥進行AIV的調查，結果於2件鴨的檢體中分離到H1N1亞型AIV，鴨的分離率為0.19% (2/1049)，鴿子為0% (0/78)，寵物鳥為 0% (0/87)。由部分NP基因的序列推論，本次所分離到的毒株，與鳥源的H1N1亞型AIV較為接近，與豬或人源的H1N1則較為疏遠。

Susceptibility to infection with avian influenza virus (AIV) was studied in healthy and immunodepressed pigeons, which were treated with the immunosuppressant cyclophosphamide (CY) before infection. Two subtypes of lowly pathogenic (LP) AIV (A/chicken/Taiwan/1209/03 H5N2 and A/chicken/Taiwan/3152/03 H6N1) were inoculated via oculonasal route. Nested reverse transcriptase-polymerase chain reaction (nested RT-PCR) and virus isolation were used as detective methods. The CY-treated and -untreated pigeons in both experiments did not shed viruses or become antibody positive throughout the 21-day observation period. All pigeons were negative for AIV RNA when trachea, lung, pancreas, spleen, kidney and rectum tissues were tested. The negative results were also obtained in noninoculated chickens which placed as contact controls in H6N1 AIV experiments. Therefore, it was concluded that the pigeons were resistant to infection with these LPAIV, and did not serve as a transmissible host, even in immune dysfunction. Following oculonasal inoculation with LPAIV (H5N2) in broiler chickens, virus was detected in trachea and rectum on 5 and 7 days postinoculation (DPI) by nested RT-PCR. AIV was also detected in myocardium and gizzard on 5 DPI, but rarely detected in brain and marrow on 5 or 7 DPI. No AI virus was prsent in breast, wing and thigh meat. The AI survey for 994 ducks, 71 pigeons and 80 pet birds in Taiwan was conducted from October 2004 to April 2005, we isolated two H1N1 AIVS from ducks. The isolation rates were 0.19% (2/1049), 0% (0/78) and 0% (0/87) in ducks, pigeons and pet birds, respectively. The partial NP genes of this two isolates were found to be closer to H1N1 avian viruses and to be different from H1N1 swine and human viruses.

目次 I
表次 V
圖次 VI
中文摘要 VII
英文摘要 VIII
第一章 緒言 1
第二章 文獻回顧 2
第一節 歷史背景 2
1-1 野鳥 3
1-2 走鳥類 4
1-3 寵物鳥 5
1-4 家禽 5
1-5 其它動物 9
第二節 病毒性質 9
2-1 家禽流行性感冒病毒的性質、型態構造及功能 9
2-2 家禽流行性感冒病毒的複製 11
2-3 家禽流行性感冒病毒生存能力 12
第三節 病徵 13
第四節 流行病學 14
第五節 家禽流行性感冒之診斷 15
5-1 直接偵測病毒蛋白或核酸 15
5-2 病毒分離 16
5-3 血清學 16
5-4 實驗室病原性評估 17
5-5 區別診斷 17
第六節 家禽流行性感冒疫苗使用現況 18
第七節 環磷醯胺 19
7-1 基本介紹 20
7-2 對禽鳥類之影響 21
7-3 對疾病感染之影響 21
第三章 材料與方法 23
第一節 鴿子對低病原性禽流感H5N2與H6N1亞型病毒之感受性 23
1-1 動物來源I 23
1-2 病毒來源I 23
1-3 試驗設計I 23
1-4 動物來源II 25
1-5 病毒來源II 25
1-6 試驗設計II 25
1-7 血球凝集試驗 26
1-8 血球凝集抑制試驗 26
1-9 病毒核酸萃取 27
1-10 反轉錄聚合酶連鎖反應 28
1-11 巢式反轉錄聚合酶連鎖反應 (nested RT-PCR) 29
1-12 nested RT-PCR增幅產物分析 29
1-13 病毒分離與同定 30
第二節 H5N2亞型低病原性禽流感病毒在人工感染肉雞臟器之分佈情形 30
2-1 實驗材料 30
2-2 試驗設計 31
2-3 核酸萃取 31
2-4 nested RT-PCR 31
第三節 調查台灣地區鴨、鴿及寵物鳥禽流感之感染情形 32
3-1 檢體來源 32
3-2 實驗方法 32
3-3 增幅NP部分基因 32
3-4 TA cloning 32
3-5 轉形反應 33
3-6 菌落篩選 33
3-7 菌落之聚合酶連鎖反應 34
3-8 基因序列分析 34
第四章 結果 35
第一節 鴿子對低病原性禽流感H5N2與H6N1亞型病毒之感受性 35
1-1 Cy對鴿與雞免疫系統之影響 35
1-2 鴿子人工感染H5N2台灣禽流感分離株之結果 35
1-3 鴿子人工感染H6N1台灣禽流感分離株之結果 36
1-4 雞人工感染H6N1台灣禽流感分離株之結果 37
1-5 陰性對照組及免疫抑制對照組 37
第二節 H5N2亞型低病原性禽流感病毒在人工感染肉雞臟器之分佈情形 37
2-1 nested RT-PCR結果 37
2-2 病毒分離結果 38
第三節 台灣地區鴨、鴿及寵物鳥之家禽流行性感冒調查結果
(93年10月~94年4月) 38
3-1 nested RT-PCR結果 38
3-2 病毒分離結果 39
3-3 鴨分離株之部分NP基因序列分析 39
第五章 討論 53
第六章 參考文獻 62
附錄 75

1.王靖雯。2002。台灣家禽流行性感冒病毒H6亞型之分子演化與毒力分析。碩士論文。台北：國立台灣大學獸醫學研究所。

2.李敏旭。1999。應用反轉錄聚合酶鏈反應診斷家禽流行性感冒病毒及區分血清亞型。碩士論文。台中：國立中興大學獸醫學研究所。

3.李照陽。2002。台灣地區鴿傳染病血清學調查及病原分離。碩士論文。台北：國立台灣大學獸醫學研究所。

4.呂榮修。1997。禽病診斷彩色圖譜。立麥登，台北，1-6。

5.廖宜真。2004。台灣地區跨越宿主傳播的人流行性感冒病毒偵測與分子及血清流行病學探究。碩士論文。台北：國立台灣大學流行病學研究所。

6.嚴慧玲。2000。台灣地區鳥類流行性感冒病毒跨越宿主傳播之流行病學研究。碩士論文。台北：國立台灣大學流行病學研究所。

7.Alexander, D. J. 2000. A review of avian influenza in different bird species. Vet Microbiol 74:3-13.

8.Baigent, S. J., and J. W. McCauley. 2003. Influenza type A in humans, mammals and birds: determinants of virus virulence, host-range and interspecies transmission. Bioessays 25:657-71.

9.Boivin, G., S. Cote, P. Dery, G. De Serres, and M. G. Bergeron. 2004. Multiplex real-time PCR assay for detection of influenza and human respiratory syncytial viruses. J Clin Microbiol 42:45-51.

10.Brown, C. C., H. J. Olander, and D. A. Senne. 1992. A pathogenesis study of highly pathogenic avian influenza virus H5N2 in chickens, using immunohistochemistry. J Comp Pathol 107:341-8.

11.Brugh, M. 1988. Highly pathogenic virus recovered from chickens infected with mildly pathogenic 1986 isolates of H5N2 avian influenza virus. Avian Dis 32:695-703.

12.Callan, R. J., G. Early, H. Kida, and V. S. Hinshaw. 1995. The appearance of H3 influenza viruses in seals. J Gen Virol 76 (Pt 1):199-203.

13.Cameron, K. R., V. Gregory, J. Banks, I. H. Brown, D. J. Alexander, A. J. Hay, and Y. P. Lin. 2000. H9N2 subtype influenza A viruses in poultry in pakistan are closely related to the H9N2 viruses responsible for human infection in Hong Kong. Virology 278:36-41.

14.Capua, I., and D. J. Alexander. 2002. Avian influenza and human health. Acta Trop 83:1-6.

15.Capua, I., and D. J. Alexander. 2004. Avian influenza: recent developments. Avian Pathol 33:393-404.

16.Capua, I., and S. Marangon. 2004. Vaccination for avian influenza in Asia. Vaccine 22:4137-8.

17.Capua, l., and S. Marangon. 2003. The use of vaccination as an option for the control of avian influenza. Office international des epizooties International committee:18-23.

18.Cattoli, G., A. Drago, S. Maniero, A. Toffan, E. Bertoli, S. Fassina, C. Terregino, C. Robbi, G. Vicenzoni, and I. Capua. 2004. Comparison of three rapid detection systems for type A influenza virus on tracheal swabs of experimentally and naturally infected birds. Avian Pathol 33:432-7.

19.Chan, P. K. 2002. Outbreak of avian influenza A(H5N1) virus infection in Hong Kong in 1997. Clin Infect Dis 34 Suppl 2:S58-64.

20.Chen, G. W., C. C. Yang, K. C. Tsao, C. G. Huang, L. A. Lee, W. Z. Yang, Y. L. Huang, T. Y. Lin, and S. R. Shih. 2004. Influenza A virus PB1-F2 gene in recent Taiwanese isolates. Emerg Infect Dis 10:630-6.

21.Coignoul, F., and H. Vindevogel. 1980. Cellular changes in the Bursa of Fabricius and thymus of cyclophosphamide-treated pigeons. J Comp Pathol 90:395-400.

22.Coppoc, G. L. 1995. Chemotherapy of neoplastic diseases, Ames, Iowa: Iowa State Press.

23.Corrier, D. E. 1991. Mycotoxicosis: mechanisms of immunosuppression. Vet Immunol Immunopathol 30:73-87.

24.Davison, S., A. F. Ziegler, and R. J. Eckroade. 1998. Comparison of an antigen-capture enzyme immunoassay with virus isolation for avian influenza from field samples. Avian Dis 42:791-5.

25.Dohms, J. E., and A. Metz. 1991. Stress--mechanisms of immunosuppression. Vet Immunol Immunopathol 30:89-109.

26.Dovc, A., O. Zorman-Rojs, A. Vergles Rataj, V. Bole-Hribovsek, U. Krapez, and M. Dobeic. 2004. Health status of free-living pigeons (Columba livia domestica) in the city of Ljubljana. Acta Vet Hung 52:219-26.

27.Dowdle, W. R., and G. C. Schild. 1975. Laboratory propagation of human influenza viruses, experimental host range and isolation from clinical materials., Academic Press: NY.

28.Dunn, P. A., E. A. Wallner-Pendleton, H. Lu, D. P. Shaw, D. Kradel, D. J. Henzler, P. Miller, D. W. Key, M. Ruano, and S. Davison. 2003. Summary of the 2001-02 Pennsylvania H7N2 low pathogenicity avian influenza outbreak in meat type chickens. Avian Dis 47:812-6.

29.Ferguson, N. M., A. P. Galvani, and R. M. Bush. 2003. Ecological and immunological determinants of influenza evolution. Nature 422:428-33.

30.Fiszon, B., C. Hannoun, A. Garcia-Sastre, E. Villar, and J. A. Cabezas. 1989. Comparison of biological and physical properties of human and animal A(H1N1) influenza viruses. Res Virol 140:395-404.

31.Fouchier, R. A., V. Munster, A. Wallensten, T. M. Bestebroer, S. Herfst, D. Smith, G. F. Rimmelzwaan, B. Olsen, and A. D. Osterhaus. 2005. Characterization of a novel influenza A virus hemagglutinin subtype (H16) obtained from black-headed gulls. J Virol 79:2814-22.

32.Fouchier, R. A., B. Olsen, T. M. Bestebroer, S. Herfst, L. van der Kemp, G. F. Rimmelzwaan, and A. D. Osterhaus. 2003. Influenza A virus surveillance in wild birds in Northern Europe in 1999 and 2000. Avian Dis 47:857-60.

33.Fulton, R. M., W. M. Reed, H. L. Thacker, and D. B. DeNicola. 1996. Cyclophosphamide (Cytoxan)-induced hematologic alterations in specific-pathogen-free chickens. Avian Dis 40:1-12.

34.Garten, W., F. X. Bosch, D. Linder, R. Rott, and H. D. Klenk. 1981. Proteolytic activation of the influenza virus hemagglutinin: The structure of the cleavage site and the enzymes involved in cleavage. Virology 115:361-74.

35.Gorman, O. T., W. J. Bean, Y. Kawaoka, and R. G. Webster. 1990. Evolution of the nucleoprotein gene of influenza A virus. J Virol 64:1487-97.

36.Govorkova, E. A., J. E. Rehg, S. Krauss, H. L. Yen, Y. Guan, M. Peiris, T. D. Nguyen, T. H. Hanh, P. Puthavathana, H. T. Long, C. Buranathai, W. Lim, R. G. Webster, and E. Hoffmann. 2005. Lethality to ferrets of H5N1 influenza viruses isolated from humans and poultry in 2004. J Virol 79:2191-8.

37.Gregory, V., M. Bennett, Y. Thomas, L. Kaiser, W. Wunderli, H. Matter, A. Hay, and Y. P. Lin. 2003. Human infection by a swine influenza A (H1N1) virus in Switzerland. Arch Virol 148:793-802.

38.Guarner, J., W. J. Shieh, J. Dawson, K. Subbarao, M. Shaw, T. Ferebee, T. Morken, K. B. Nolte, A. Freifeld, N. Cox, and S. R. Zaki. 2000. Immunohistochemical and in situ hybridization studies of influenza A virus infection in human lungs. Am J Clin Pathol 114:227-33.

39.Hay, A. J., B. Lomniczi, A. R. Bellamy, and J. J. Skehel. 1977. Transcription of the influenza virus genome, vol. 83.

40.Hoffmann, E., J. Stech, Y. Guan, R. G. Webster, and D. R. Perez. 2001. Universal primer set for the full-length amplification of all influenza A viruses. Arch Virol 146:2275-89.

41.Hooper, P. T., G. W. Russell, P. W. Selleck, and W. L. Stanislawek. 1995. Observations on the relationship in chickens between the virulence of some avian influenza viruses and their pathogenicity for various organs. Avian Dis 39:458-64.

42.Huang, R. T., R. Rott, and H. D. Klenk. 1981. Influenza viruses cause hemolysis and fusion of cells. Virology 110:243-7.

43.Islam, A. F., C. W. Wong, S. W. Walkden-Brown, I. G. Colditz, K. E. Arzey, and P. J. Groves. 2002. Immunosuppressive effects of Marek''s disease virus (MDV) and herpesvirus of turkeys (HVT) in broiler chickens and the protective effect of HVT vaccination against MDV challenge. Avian Pathol 31:449-61.

44.Kaleta, E. F., and A. Honicke. 2004. Review of the literature on avian influenza A viruses in pigeons and experimental studies on the susceptibility of domestic pigeons to influenza A viruses of the haemagglutinin subtype H7. Dtsch Tierarztl Wochenschr 111:467-72.

45.Karasin, A. I., K. West, S. Carman, and C. W. Olsen. 2004. Characterization of avian H3N3 and H1N1 influenza A viruses isolated from pigs in Canada. J Clin Microbiol 42:4349-54.

46.Kewalramani, V. N., A. T. Panganiban, and M. Emerman. 1992. Spleen necrosis virus, an avian immunosuppressive retrovirus, shares a receptor with the type D simian retroviruses. J Virol 66:3026-31.

47.Krug, R. M. 2003. The potential use of influenza virus as an agent for bioterrorism. Antiviral Res 57:147-50.

48.Lamb, R. A., and R. M. Krug. 2001. Orthomyxoviridae: The viruses and their replication. Linppincott Williams & Wilkins, Philadelphia, PA.

49.Lamb, R. A., R. M. Krug. 1996. Orthomyxoviridae: The viruses and their replication, Lippincott-Raven: NY.

50.Latshaw, J. D. 1991. Nutrition--mechanisms of immunosuppression. Vet Immunol Immunopathol 30:111-20.

51.Lee, C. W., and D. L. Suarez. 2004. Application of real-time RT-PCR for the quantitation and competitive replication study of H5 and H7 subtype avian influenza virus. J Virol Methods 119:151-8.

52.Lee, M. S., P. C. Chang, J. H. Shien, M. C. Cheng, and H. K. Shieh. 2001. Identification and subtyping of avian influenza viruses by reverse transcription-PCR. J Virol Methods 97:13-22.

53.Liu, M., Y. Guan, M. Peiris, S. He, R. J. Webby, D. Perez, and R. G. Webster. 2003. The quest of influenza A viruses for new hosts. Avian Dis 47:849-56.

54.Lu, B. L., R. G. Webster, and V. S. Hinshaw. 1982. Failure to detect hemagglutination-inhibiting antibodies with intact avian influenza virions. Infect Immun 38:530-5.

55.Lu, X., D. Cho, H. Hall, T. Rowe, H. Sung, W. Kim, C. Kang, I. Mo, N. Cox, A. Klimov, and J. Katz. 2003. Pathogenicity and antigenicity of a new influenza A (H5N1) virus isolated from duck meat. J Med Virol 69:553-9.

56.Lu, X. H., D. Cho, H. Hall, T. Rowe, I. P. Mo, H. W. Sung, W. J. Kim, C. Kang, N. Cox, A. Klimov, and J. M. Katz. 2003. Pathogenesis of and immunity to a new influenza A (H5N1) virus isolated from duck meat. Avian Dis 47:1135-40.

57.Ludwig, S., A. Haustein, E. F. Kaleta, and C. Scholtissek. 1994. Recent influenza A (H1N1) infections of pigs and turkeys in northern Europe. Virology 202:281-6.
58.Lutticken, D. 1997. Viral diseases of the immune system and strategies to control infectious bursal disease by vaccination. Acta Vet Hung 45:239-49.

59.Markowski-Grimsrud, C. J., and K. A. Schat. 2003. Infection with chicken anaemia virus impairs the generation of pathogen-specific cytotoxic T lymphocytes. Immunology 109:283-94.

60.Marozin, S., V. Gregory, K. Cameron, M. Bennett, M. Valette, M. Aymard, E. Foni, G. Barigazzi, Y. Lin, and A. Hay. 2002. Antigenic and genetic diversity among swine influenza A H1N1 and H1N2 viruses in Europe. J Gen Virol 83:735-45.

61.Meulemans, G., M. C. Carlier, M. Gonze, and P. Petit. 1987. Comparison of hemagglutination-inhibition, agar gel precipitin, and enzyme-linked immunosorbent assay for measuring antibodies against influenza viruses in chickens. Avian Dis 31:560-3.

62.Mo, I. P., M. Brugh, O. J. Fletcher, G. N. Rowland, and D. E. Swayne. 1997. Comparative pathology of chickens experimentally inoculated with avian influenza viruses of low and high pathogenicity. Avian Dis 41:125-36.

63.Myers, T. J., M. D. Rhorer, and J. Clifford. 2003. USDA options for regulatory changes to enhance the prevention and control of avian influenza. Avian Dis 47:982-7.

64.Naeem, K., T. Niazi, S. A. Malik, and A. H. Cheema. 1995. Immunosuppressive potential and pathogenicity of an avian adenovirus isolate involved in hydropericardium syndrome in broilers. Avian Dis 39:723-8.

65.Nakamura, K., Y. Imada, and F. ABE. 1987. Effect of cyclophosphamide on infectious produced by Escherichia coli of high and low virulence in chickens. Avian Pathology 16:237-252.

66.Nakamura, K., T. Shoyama, M. Mase, T. Imada, and M. Yamada. 2003. Reproduction of hydropericardium syndrome in three-week-old cyclophosphamide-treated specific-pathogen-free chickens by adenoviruses from inclusion body hepatitis. Avian Dis 47:169-74.

67.Nakamura, R. M., and B. C. Easterday. 1967. Serological studies of influenza in animals. Bull World Health Organ 37:559-67.

68.Narayan, O., G. Lang, and B. T. Rouse. 1969. A new influenza A virus infection in turkeys. IV. Experimental susceptibility of domestic birds to virus strain turkey-Ontario 7732-1966. Arch Gesamte Virusforsch 26:149-65.

69.Panigrahy, B., D. A. Senne, and J. C. Pedersen. 2002. Avian influenza virus subtypes inside and outside the live bird markets, 1993-2000: a spatial and temporal relationship. Avian Dis 46:298-307.

70.Panigrahy, B., D. A. Senne, J. C. Pedersen, A. L. Shafer, and J. E. Pearson. 1996. Susceptibility of pigeons to avian influenza. Avian Dis 40:600-4.

71.Panigrahy, J. E. 1987. Presented at the Proc 57th avian disease conference, Atlantic City, N. J.

72.Pasick, J., H. Weingartl, A. Clavijo, J. Riva, H. Kehler, K. Handel, E. Watkins, and K. Hills. 2003. Characterization of avian influenza virus isolates submitted to the National Centre for Foreign Animal Disease between 1997 and 2001. Avian Dis 47:1208-13.

73.Perdue, M. L. 2003. Molecular diagnostics in an insecure world. Avian Dis 47:1063-8.

74.Perkins, L. E., and D. E. Swayne. 2003. Comparative susceptibility of selected avian and mammalian species to a Hong Kong-origin H5N1 high-pathogenicity avian influenza virus. Avian Dis 47:956-67.

75.Perkins, L. E., and D. E. Swayne. 2002. Pathogenicity of a Hong Kong-origin H5N1 highly pathogenic avian influenza virus for emus, geese, ducks, and pigeons. Avian Dis 46:53-63.

76.Pharo, H. J. 2003. The impact of new epidemiological information on a risk analysis for the introduction of avian influenza viruses in imported poultry meat. Avian Dis 47:988-95.

77.Pope, C. R. 1991. Chicken anemia agent. Vet Immunol Immunopathol 30:51-65.

78.Purchase, H. G. 1989. Practical application of nucleic acid techniques to avian disease problems. Avian Dis 33:609-14.

79.Reisen, W. K., R. E. Chiles, E. N. Green, Y. Fang, F. Mahmood, V. M. Martinez, and T. Laver. 2003. Effects of immunosuppression on encephalitis virus infection in the house finch, Carpodacus mexicanus. J Med Entomol 40:206-14.

80.Reisen, W. K., R. E. Chiles, V. M. Martinez, Y. Fang, and E. N. Green. 2004. Encephalitis virus persistence in California birds: experimental infections in mourning doves (Zenaidura macroura). J Med Entomol 41:462-6.

81.Ruigrok, R. W., E. A. Hewat, and R. H. Wade. 1992. Low pH deforms the influenza virus envelope. J Gen Virol 73 (Pt 4):995-8.

82.Saif, Y. M. 1991. Immunosuppression induced by infectious bursal disease virus. Vet Immunol Immunopathol 30:45-50.

83.Schoch-Spana, M. 2000. Implications of pandemic influenza for bioterrorism response. Clin Infect Dis 31:1409-13.

84.Scholtissek, C., S. Ludwig, and W. M. Fitch. 1993. Analysis of influenza A virus nucleoproteins for the assessment of molecular genetic mechanisms leading to new phylogenetic virus lineages. Arch Virol 131:237-50.

85.Schultz, U., W. M. Fitch, S. Ludwig, J. Mandler, and C. Scholtissek. 1991. Evolution of pig influenza viruses. Virology 183:61-73.

86.Senne, D. A., J. E. Pearson, L. D. Miller, and G. A. Gustafson. 1983. Virus isolations from pet birds submitted for importation into the United States. Avian Dis 27:731-44.

87.Slemons, R. D., and B. C. Easterday. 1972. Host response differences among 5 avian species to an influenzavirus--A-turkey-Ontario-7732-66 (Hav5N?). Bull World Health Organ 47:521-5.

88.Slemons, R. D., and D. E. Swayne. 1990. Replication of a waterfowl-origin influenza virus in the kidney and intestine of chickens. Avian Dis 34:277-84.

89.Sominina, A. A., T. P. Lisok, and N. B. Rumel. 1977. Improved methods of influenza virus propagation. II. Characteristics of cell culture and allantoic virus preparations. Acta Virol 21:241-5.

90.Spackman, E., D. A. Senne, L. L. Bulaga, T. J. Myers, M. L. Perdue, L. P. Garber, K. Lohman, L. T. Daum, and D. L. Suarez. 2003. Development of real-time RT-PCR for the detection of avian influenza virus. Avian Dis 47:1079-82.

91.Spackman, E., D. A. Senne, L. L. Bulaga, S. Trock, and D. L. Suarez. 2003. Development of multiplex real-time RT-PCR as a diagnostic tool for avian influenza. Avian Dis 47:1087-90.

92.Spackman, E., D. A. Senne, T. J. Myers, L. L. Bulaga, L. P. Garber, M. L. Perdue, K. Lohman, L. T. Daum, and D. L. Suarez. 2002. Development of a real-time reverse transcriptase PCR assay for type A influenza virus and the avian H5 and H7 hemagglutinin subtypes. J Clin Microbiol 40:3256-60.

93.Stallknecht, D. E., M. T. Kearney, S. M. Shane, and P. J. Zwank. 1990. Effects of pH, temperature, and salinity on persistence of avian influenza viruses in water. Avian Dis 34:412-8.

94.Starick, E., and O. Werner. 2003. Detection of H7 avian influenza virus directly from poultry specimens. Avian Dis 47:1187-9.

95.Suarez, D. L. 2000. Evolution of avian influenza viruses. Vet Microbiol 74:15-27.

96.Suarez, D. L., and M. L. Perdue. 1998. Multiple alignment comparison of the non-structural genes of influenza A viruses. Virus Res 54:59-69.

97.Sugimura, T., Y. Murakami, and T. Ogawa. 2000. The susceptibility of culture cells to avian influenza viruses. J Vet Med Sci 62:659-60.

98.Suss, J., J. Schafer, H. Sinnecker, and R. G. Webster. 1994. Influenza virus subtypes in aquatic birds of eastern Germany. Arch Virol 135:101-14.

99.Swayne, D. E. 1997. Pathobiology of H5N2 Mexican avian influenza virus infections of chickens. Vet Pathol 34:557-67.

100.Swayne, D. E., and J. R. Beck. 2005. Experimental study to determine if low-pathogenicity and high-pathogenicity avian influenza viruses can be present in chicken breast and thigh meat following intranasal virus inoculation. Avian Dis 49:81-85.

101.Swayne, D. E., and D. A. Halvorson. 2003. Influenza. Iowa State Press, Ames, Iowa.

102.Swayne, D. E., and D. J. King. 2003. Avian influenza and Newcastle disease. J Am Vet Med Assoc 222:1534-40.

103.Taubenberger, J. K., A. H. Reid, A. E. Krafft, K. E. Bijwaard, and T. G. Fanning. 1997. Initial genetic characterization of the 1918 "Spanish" influenza virus. Science 275:1793-6.

104.Trust, K. A., J. R. Fowles, M. J. Hooper, and A. Fairbrother. 1994. Cyclophosphamide effects on immune function of European starlings. J Wildl Dis 30:328-34.

105.Tumpey, T. M., D. L. Suarez, L. E. Perkins, D. A. Senne, J. Lee, Y. J. Lee, I. P. Mo, H. W. Sung, and D. E. Swayne. 2003. Evaluation of a high-pathogenicity H5N1 avian influenza A virus isolated from duck meat. Avian Dis 47:951-5.

106.Tumpey, T. M., D. L. Suarez, L. E. Perkins, D. A. Senne, J. G. Lee, Y. J. Lee, I. P. Mo, H. W. Sung, and D. E. Swayne. 2002. Characterization of a highly pathogenic H5N1 avian influenza A virus isolated from duck meat. J Virol 76:6344-55.

107.Turek, R., M. Gresikova, and B. Tumova. 1984. Isolation of influenza A virus and paramyxoviruses from sentinel domestic ducks. Acta Virol 28:156-8.

108.Vindevogel, H., P. P. Pastoret, and G. Burtonboy. 1980. Pigeon herpes infection: excretion and re-excretion of virus after experimental infection. J Comp Pathol 90:401-8.

109.Wang, C. W., and C. H. Wang. 2003. Experimental selection of virus derivatives with variations in virulence from a single low-pathogenicity H6N1 avian influenza virus field isolate. Avian Dis 47:1416-22.

110.Weir, E., T. Wong, and I. Gemmill. 2004. Avian influenza outbreak: update. Cmaj 170:785-6.

111.WHO. 2002. WHO manual on animal influenza diagnosis and surveillance. Global influenza programme, World Health Organization, Department of communicable diseases surveillance and control, Switzerland.

112.Yen, H. L., M. C. Cheng, J. L. Liu, C. L. Kao, S. R. Shih, N. J. Cox, R. G. Webster, and C. C. King. 2001. Influenza surveillance in poultry market and its inter-species transmission in Taiwan. International Congress Series 1219:201-11.

113.Yuen, K. Y., P. K. Chan, M. Peiris, D. N. Tsang, T. L. Que, K. F. Shortridge, P. T. Cheung, W. K. To, E. T. Ho, R. Sung, and A. F. Cheng. 1998. Clinical features and rapid viral diagnosis of human disease associated with avian influenza A H5N1 virus. Lancet 351:467-71.