臺灣博碩士論文加值系統

溶血性曼哈米亞桿菌 (Mannheimia haemolytica) 屬於巴氏桿菌科，主要會造成反芻類動物急性纖維素性胸膜肺炎，導致感染動物迅速死亡，藉由接種疫苗來達到預防是目前研究的重點，但目前缺乏有效的疫苗，因此需開發新一代的疫苗來降低經濟上的損失。由前人研究中已知外膜蛋白(GS60)為很重要的抗原，而白血球毒素(LKT)則是增加肺部病變元兇，本研究以原核系統表現重組蛋白並分析其抗原性。首先由牛羊篩選M. haemolytica本土株，試製不活化菌苗，並添加可被免疫後仔牛血清所辨識之重組蛋白rGS60，與重組毒素蛋白rLKT，再和水包油包水佐劑混合試製成不同疫苗: M. h、rGS60、M. h+ rGS60、rLKT、M. h+rLKT。於小鼠的攻毒試驗，免疫組均有60-80%的存活率，而對照組則全部死亡；小鼠補強免疫抗體試驗中，免疫組與對照組相較，均有顯著性之差異(p<0.01)；由抗體亞型分析，所有菌苗均以產生IgG2a抗體為主；於小鼠細胞性增生試驗中，免疫組均可刺激細胞增生。於仔羊實驗中，免疫rGS60組抗體力價可持續至12週之久，且可刺激細胞增生與對照組有顯著之差異(p<0.01)。免疫rGS60及rLKT組均有表現IFN-γ；抗體力價與細胞增生試驗rLKT較rGS60有顯著差異(p<0.01)。綜合上述結果，証實兩種重組蛋白之抗原性佳，具有開發為新型疫苗與佐劑之潛力。

Mannheimia (Pasteurella) haemolytica is a member of the Pasteurellaceae family, which is the major cause of acute fibrinous pneumonia in ruminants. The mortality is high and vaccination is the major strategy for controlling this disease. There is no effect vaccine to protect pneumonic pasteurellosis. It is urgency to develop new vaccine to prevent M. haemolytica infection and reduce the economical loss. The previous studies indicated that outer membrane proteins GS60 are the most important antigens and the leukotoxin (LKT) plays a major role in lung injury. Our objectives are to clone and express GS60 and LKT of M. haemolytica by the prokaryotic expression system and characterize their antigenicities. The vaccines (M. h, M. h+rGS60, rGS60, M. h+rLKT, rLKT) made of inactivated wild-type M. haemolytica and recombinant proteins (rGS60, rLKT) with w/o/w adjuvant. The survival rate of protection efficacy in immunized mice was 60% to 80%, while control mice were all dead. The antibody titer of immunized mice was significantly higher than control group (p&lt;0.01) and the major subtype was IgG2a. The T-cell proliferation index of immunized groups was significantly higher than control group (p&lt;0.01) in mice and goat. Moreover, the antibody titer of immunized rGS60 goat was significantly higher than control group (p&lt;0.01) and continued at the 12th week. The IFN-γ gene expression was detected in vaccinated (rLKT and rGS60) goats. Furthermore the rLKT antibody titer and T-cell proliferation index was significantly higher than rGS60 group (p&lt;0.01) at 4 weeks. In summary, these expressed proteins could induce humoral and cellular immune response used as a potential immunogen or adjuvant to develop new vaccines against M. haemolytica.

目 錄
中文摘要............................................ Ⅰ
Abstract........................................... Ⅱ
謝誌............................................... Ⅳ
目錄............................................... Ⅴ
圖表目錄............................................ Ⅸ
第1章 前言....................................... 1
第2章 文獻回顧................................... 4
2.1 巴氏桿菌症................................. 4
2.1.1 巴氏桿菌症造成經濟損失之重要性............... 4
2.1.2 巴氏桿菌疾病之成因......................... 4
2.1.3 巴氏桿菌症臨床症狀和病變.................... 5
2.2 Mannheimia haemolytica (溶血性曼哈米亞桿菌). 6
2.2.1 M. haemolytica之生物分類.................. 6
2.2.2 M. haemolytica之型態與生化特性............. 7
2.2.3 M. haemolytica之致病機轉.................. 7
2.2.4 M. haemolytica之毒力因子.................. 8
2.2.4.1 外膜脂蛋白(outer membrane lipoprotein).... 9
2.2.4.2 白血球毒素 (lukotoxin).................... 10
2.2.4.3 白血球毒素造成細胞凋亡和壞死之機轉........... 12
2.3 巴氏桿菌肺炎症之治療與預防.................. 14
2.4 巴氏桿菌疫苗開發現況........................ 15
第3章 材料與方法................................. 16
3.1 M. haemolytica之生化鑑定.................. 16
3.2 M. haemolytica之培養..................... 16
3.3 M. haemolytica lps A與gs60之選殖與定序.... 16
3.3.1 M. haemolytica genomic DNA之萃取......... 16
3.3.2 聚合酶連鎖反應............................ 17
3.3.2.1 快速鑑定曼哈米亞桿菌之lps A引子............. 17
3.3.2.2 M. haemolytica gs60之選殖 ............... 17
3.3.3 DNA之膠體電泳法............................ 18
3.3.4 PCR反應產物之純化.......................... 18
3.3.5 PCR反應產物之定序.......................... 18
3.4 以原核表現系統表現gs60之重組蛋白質.......... 19
3.4.1 gs60基因之製備............................ 19
3.4.2 表現載體之製備........................... 19
3.4.3 gs60與表現載體接合反應..................... 19
3.4.4 勝任細胞之製備............................ 20
3.4.5 轉形作用................................. 20
3.4.6 小量質體DNA之萃取......................... 20
3.5 重組蛋白質rGS60之表現與分析................ 21
3.5.1 重組蛋白rGs60之表現....................... 21
3.5.2 蛋白質電泳................................ 21
3.5.2.1 蛋白質之製備……………………………........... 21
3.5.2.2 聚丙烯醯胺膠體之製備…………................. 21
3.5.2.3 蛋白質之定量……………………................. 22
3.5.3 重組蛋白質之抗原性分析-西方墨點法............. 22
3.6 疫苗之製備................................ 23
3.6.1 M. haemolytca 之定量與不活化............... 23
3.6.2 重組蛋白之培養與不活化...................... 23
3.6.3 疫苗之調配................................ 23
3.7 動物試驗................................. 24
3.7.1 小鼠半致死劑量試驗......................... 24
3.7.2 小鼠效力試驗.............................. 24
3.7.3 仔羊效力試驗............................... 25
3.8 免疫反應之分析............................ 25
3.8.1 酵素連結免疫吸附法......................... 25
3.8.2 IgG亞型抗體 (subclass) 之分析............ 26
3.8.3 免疫細胞增生試驗.......................... 26
3.8.3.1 小鼠免疫................................. 26
3.8.3.2 脾臟細胞之分離............................ 26
3.8.3.3 周邊血液單核球細胞之抽取................... 27
3.8.3.4 細胞計數................................. 27
3.8.3.5 抗原刺激................................. 27
3.8.3.6 MTT assay............................... 27
3.8.3.7 細胞增生指數 SI值計算...................... 28
3.9 細胞毒殺試驗.............................. 28
3.9.1 多形核白血球之抽取......................... 28
3.9.2 分泌型白血球毒素之製備..................... 28
3.9.3 白血球毒素之抗原性分析..................... 29
3.9.4 白血球毒素之毒殺作用....................... 29
3.9.5 MTT assay............................... 29
3.9.6 細胞毒殺能力之計算.......................... 29
3.9.7 重組白血球毒素抗體之中和能力.................. 29
3.10 流式細胞儀分析分泌型白血球毒素之活性.......... 29
3.10.1 多形核白血球之製備......................... 30
3.10.2 抗體染色................................. 30
3.10.3 白血球毒素刺激CD11a/CD18之表現............. 30
3.11 以RT-PCR方法檢測周邊血液單核球細胞IFN-γ的產生量. 30
3.11.1 周邊血液單核球細胞之製備................... 30
3.11.2 RNA抽取.................................. 30
3.11.3 反轉錄聚合酶連鎖反應....................... 31
3.11.4 IFN-γ與β-actin之分析....................... 32
3.12 統計分析.................................. 33
第4章 結果..................................... 34
4.1 M. haemolytica之生化試驗................... 34
4.2 M. haemolytica之PCR鑑定.................. 34
4.2.1 M. haemolytica lpsA之選殖與定序........... 34
4.3 M. haemolytica gs60 之選殖、定序與胺基酸比對. 35
4.4 重組蛋白質rGS60 之表現與分析............... 35
4.5 動物試驗.................................. 35
4.5.1 小鼠半致死劑量(LD50) ...................... 35
4.5.2 rGS60效力試驗............................. 36
4.5.2.1 小鼠效力試驗............................... 36
4.5.2.2 小鼠攻毒效力試驗........................... 36
4.5.2.3 仔羊效力試驗............................... 36
4.5.3 rLKT效力試驗............................... 37
4.5.3.1 小鼠效力試驗................................ 37
4.5.3.2 小鼠攻毒效力試驗............................ 37
4.6 免疫反應之分析法............................ 37
4.6.1 rLKT與rGS60抗體亞型之分析.................. 37
4.6.2 rGS60小鼠細胞性免疫反應試驗........................................... 38
4.6.3 rLKT 小鼠細胞性免疫反應試驗........................................... 38
4.6.4 rGS60仔羊細胞性免疫反應試驗............................................ 38
4.6.5 rGS60與rLKT仔羊之效力試驗............................................ 38
4.6.6 rGS60與rLKT仔羊之細胞性免疫反應試驗...... 39
4.6.7 rGS60與rLKT仔羊之IFN-γ表現量分析........ 39
4.7 白血球毒素之製備與抗原性分析............. 39
4.8 牛多形核白血球CD11a/CD18之表現量........ 39
第5章 討論.................................. 65
參考文獻...................................... 71
附錄......................................... 84
Appendix 1.The positive percentage of bovine diseases in Taiwan........... 84
Appendix 2.The positive percentage of bovine diseases cross infection in Taiwan.. 85
Appendix 3.M. haemolytica leukotoxin gene cluster. 86
Appendix 4.Leukocyte signaling pathway triggered by LKT and LPS.............. 87
Appendix 5.pGEM-T Easy vector map......... 88
Appendix 6.pET-32a vector map…………………………………….............. 89
作者簡介................................ . 90

朱純燕、陳貞妗、陳銘政。水包油包水 (w/o/w) 油質佐劑對牛流行熱疫苗抗體消長之影響。台灣獸醫學雜誌33 : 211-216，2007。
吳永惠。牛出血性敗血症。牛病學。藝軒圖書出版社146-147，2000。
吳幸潔。溶血性曼哈米司桿菌(巴氏桿菌)毒力基因之選殖與表現蛋白抗原性之分析。碩士論文。國立屏東科技大學獸醫學系。台灣省。中華民國。2008。
林哲祺、林素容、朱純燕。巴氏德桿菌。獸醫臨床細菌鑑定圖譜。藝軒圖書出版社155-157，1993。
劉世賢、劉炳燦、吳永惠、廖明輝。草食動物傳染性疾病監控：應用酵素結合免疫系統檢測牛羊重要呼吸道傳染病。行政院農業委員會動植物防疫檢疫局，2004。
Al-Ghamdi GM, Ames TR, Baker JC, Walker R, Chase CC, Frank GH, Maheswaran SK. Serotyping of Mannheimia (Pasteurella) haemolytica isolates from the upper Midwest United States. J Vet Diagn Invest 12: 576-578, 2000.
Angen O, Mutters R, Caugant DA, Olsen JE, Bisgaard M. Taxonomic relationships of the (Pasteurella) haemolytica complex as evaluated by DNA-DNA hybridations and 16S rRNA sequencing with proposal of Mannheimia haemolytica. Int J Syst Bacteriol 49: 67-86, 1999a.
Angen O, Quirie M, Donachie W, Bisgaard M. Investigations on the species specificity of Mannheimia (Pasteurella) haemolytica serotyping. Vet Microbiol 65: 283-290, 1999b.
Apley M. Antimicrobial therapy of bovine respiratory disease.Vet Clin North Am Food Anim Pract 3: 549-574, 1997.
Ayalew S, Blackwood ER, Confer AW. Sequence diversity of the immunogenic outer membrane lipoprotein PlpE from Mannheimia haemolytica serotypes 1, 2, and 6. Vet Microbiol 114: 260-268, 2006.
Ayalew S, Confer AW, Payton ME, Garrels KD, Shrestha B, Ingram KR, Montelongo MA, Taylor JD. Mannheimia haemolytica chimeric protein vaccine composed of the major surface-exposed epitope of outer membrane lipoprotein PlpE and the neutralizing epitope of leukotoxin. Vaccine 38: 4955-4961, 2008.
Bateman KG. Antimicrobial drug use in cattle. In Prescott JF and Baggot JD (eds) Antimicrobial Therapy in Veterinary Medicine, 2nd edu. Ames, IA: Iowa state University Press 456-468, 1993.
Biberstein EL. Biotyping and serotyping Pasteurella haemolytica. In: Bergan T and Norris JR (eds) Methods in Microbiology, London: Academic Press 253-269, 1987.
Bingham DP, Moore R, Richards AB. Comparison of DNA: DNA homology and enzymatic activity between Pasteurella haemolytica and related species. Am J Vet Res 51: 1161-1166, 1990.
Blackall PJ, Bojesen AM, Chrisensem H, Bisgaard M. Reclassification of Pasteurella trehalosi and Bibersteimia trebhalosi gen.nov., comb.nov. Int J Syst Evol Microbiol 5: 666-674, 2007.
Breider MA, Walker RD, Hopkins FM, Schultz TW, Bowersock TL. Pulmonary lesions induced by Pasteurella haemolytica in neutrophil sufficient and neutrophil deficient calves. Can J Vet Res 52: 205-209, 1988.
Brogden KA, DeBey B, Audibert F, Lehmkuhl H, Chedid L. Protection of ruminants by Pasteurella haemolytica A1 capsular polysaccharide vaccines containing muramyl dipeptide analogs.Vaccine 17: 1677-1684, 1995.
Brogden KA, Lehmkuhl HD, Cutlip RC. Pasteurella haemolytica complicated respiratory infections in sheep and goats. Vet Res 29: 233-254, 1998.
Brown GB, Bolin SR, Frsnk DE, Roth JA. Defective function of leukocytes from cattle persistently infected with bovine viral diarrhea virus, and the influence of recombinant cytokines. Am J Vet Res 49: 1447-1451, 1991.
Brown TT Jr, Ananaba G. Effect of respiratory infections caused by bovine herpesvirus-1 or parainfluenza-3 viuvs on bovine alveolar macrophage functions. Am J Vet Res 49: 1447-1451, 1988.
Chae CH, Gentry MJ, Confer AW, Anderson GA. Resistance to host immune defense mechanisms afforded by capsular material of Pasteurella haemolytica, serotype 1. Vet Microbilo 25: 241-251, 1990.
Confer AW. Immunogens of Pasteurella. Vet Microbiol. 37: 353-368, 1993.
Confer AW, Clinkenbeard KD, Gatewood DM, Driskel BA, Montelongo M. Serum antibody responses of cattle vaccinated with partially purified native Pasteurella haemolytica leukotoxin. Vaccine 15: 1423-1429, 1997.
Confer AW, Ayalew S, Panciera RJ, Montelongo M, Whitworth LC, Hammer JD. Immunogenicity of recombinant Mannheimia haemolytica serotype 1 outer membrane protein PlpE and augmentation of a commercial vaccine. Vaccine 21: 2821-2829, 2003.
Confer AW, Ayalew S, Panciera RJ, Montelongo M, Wray JH. Recombinant Mannheimia haemolytica serotype 1 outer membrane protein PlpE enhances commercial Mannheimia haemolytica vaccine-induced resistance against serotype 6 challenge. Vaccine 24: 2248-2255, 2006.
Confer AW, Ayalew S, Montelongo M, Step DL, Wray JH, Hansen RD, Panciera RJ. Immunity of cattle following vaccination with a Mannheimia haemolytica chimeric PlpE-LKT (SAC89) protein.Vaccine 11: 1771-1776, 2009.
Cooney BJ, Lo RY. Three contiguous lipoprotein genes in Pasteurella haemolytica A1 which are homologous to a lipoprotein gene in Haemophilus influenzae type b. Infect Immun 11: 4682-4688, 1993.
Coote JG. Structural and functional relationships among the RTX toxin determinants of gram-negative bacteria. FEMS Microbiol 8: 137-161, 1992.
Corbeil LB, Arnold KF, Kimball R, Berghaus L, Gershwin LJ. Specificity of IgG and IgE antibody responses to Haemophilus somnus infection of calves. Vet Immunol Immunopathol 15: 191-199, 2006.
Davies RL, Whittam TS, Selander RK. Sequence diversity and molecular evolution of the leukotoxin (lktA) gene in bovine and ovine strains of Mannheimia (Pasteurella) haemolytica. J Bacteriol 183: 1394-1404, 2001.
Davies RL, Campbell S, Whittam TS. Mosaic structure and molecular evolution of the leukotoxin operon (lktCABD) in Mannheimia (Pasteurella) haemolytica, Mannheimia glucosida, and Pasteurella trehalosi. J Bacteriol 184: 266-277, 2002.
Davies RL, Baillie S. Cytotoxic activity of Mannheimia haemolytica strains in relation todiversity of the leukotoxin structural gene lktA. Vet. Microbiol 92: 263-279, 2003.
DeBey BM, Roth JA, Brogden KA, Cutlip RC, Stevens MG, Jones T, Briggs RE, Kluge JP. In vitro lymphocyte proliferative responses and gamma-interferon production as measures of cell-mediated immunity of cattle exposed to Pasteurella haemolytica. Can J Vet Res 4: 263-270, 1996.
Diamond G, Russell JP, Bevins CL. Inducible expression of an antibiotic peptide gene in lipopolysaccharide challenged tracheal epithelial cells. Proc Natl Acad Sci. USA 93: 5156-5160, 1996.
Donachie W. Bacteriology of bovine respiratory disease. UK Vet 3: 44-46, 1998.
Dungworth D. The respiratory system, in: Jubb K, Kennedy P, Palmer N. (Eds.). Pathology of domestic animals, 4th ed., Academic Press, San Diego 589-663, 1992.
Ewers C, Lubke-Becker A, Wieler LH. Mannheimia haemolytica and the pathogenesis of enzootic bronchopneumonia. Berl Munch Tierarztl Wochenschr 117: 97-11, 2004.
Fett T, Zecchinon L, Vanden BP, Desmecht D. Mannheimia haemolytica leukotoxin-induced cytolysis of caprine (Capra hircus) leukocytes is mediated by the CD18 subunit of b2-integrins. Microb Pathog 45: 1-6, 2008.
Filion LG, Willson PJ, Bielfeldt-Ohmamn H, Babiuk LA, Thomson RG. The possible role of stress in the induction of pneumonic pasteurellosis. Can J Comp Med 48: 268-274, 1984.
Frank GH, Smith PC. Prevalence of Pasteurella haemolytica in transported calves. Am J Vet Res 44: 981-985, 1983.
Frank GH. Pasteurellosis of cattle. London: Academic Press Limited 197-222, 1989.
Frank GH, Briggs RE (1992). Colonization of the tonsils of calves with Pasteurella haemolytica. Am J Vet Res 53: 481-484, 1992.
Frey J, Kuhnert P. RTX toxins in Pasteurellaceae. Int J Med Microbiol 292: 149-158, 2002.
Gardner BA, Northcutt SL, Dolezal HG,Gill DR, Ray FK, Morgan JB, Shearhart CW. Factors influencing profitability of feedlot steers. Animal Science Research Report Oklahoma Ag Experiment Station Oklahoma State University Stillwater USA 164-172, 1996.
Gatewood DM, Fenwick BW, Chengappa MM. Growth-condition dependent expression of Pasteurella haemolytica A1 outer membrane proteins, capsule, and leukotoxin. Vet Microbiol 3: 221-233, 1994.
Griffin D. Economic impact associated with respiratory disease in beef cattle. Vet Clin North Am Food Anim Pract 367-377, 1997.
Highlander SK, Wickersham EA, Garza O, Weinstock GM. Expression of the Pasteurella haemolytica leukotoxin is inhibited by a locus that encodes an ATP-binding cassette homolog. Infect Immun 61: 3942-3951, 1993.
Highlander SK. Molecular genetic analysis of virulence in Mannheimia (Pasteurella) haemolytica. Front Biosci 6: 1128-1150, 2001.
Hirsh DC, Zee YC. Veterinary Microbiology. Boston, MA: Blackwell Scientific Publications 135-140, 1999.
Hodgson JC, Moon GM, Quirie M, Donachie W. Association of LPS chemotype of Mannheimia (Pasteurella) haemolytica A1 with disease virulence in a model of ovine pneumonic pasteurellosis. J Endotoxin Res 1: 25-32, 2002.
Jaramillo L, Díaz F, Hernández P, Debray H, Trigo F, Mendoza G, Zenteno E. Purification and characterization of an adhesin from Pasteurella haemolytica. Glycobiology 1: 31-37, 2000.
Jeyaseelan S, Kannan MS, Briggs RE, Thumbikat P, Maheswaran SK. Mannheimia haemolytica leukotoxin activates a nonreceptor tyrosine kinase signaling cascade in bovine leukocytes, which induces biological effects. Infect Immun 69: 6131-6139, 2001.
Jeyaseelan S, Sreevatsan S, Maheswaran SK. Role of Mannheimia haemolytica leukotoxin in the pathogenesis of bovine pneumonic pasteurellosis. Anim Health Res Rev 2: 69-82, 2002.
Kathaperumal K, Kumanan V, McDonough S, Chen LH, Park SU, Moreira MA, Akey B, Huntley J, Chang CF, Chang YF. Evaluation of immune responses and protective efficacy in a goat model following immunization with a coctail of recombinant antigens and a polyprotein of Mycobacterium avium subsp. paratuberculosis. Vaccine 1: 123-35, 2009.
Kitt T. Uber eine experimentelle der Rinderseuche ahnliche Infektionskranheit, Sitzunbsber. Ges Morphol Physiol Muenchen 140-168, 1885.
Lainson FA, Murray J, Davies RC, Donachie W. Characterization of epitopes involved in the neutralization of Pasteurella haemolytica serotype A1 leukotoxin. Microbiology 142: 2499-2507, 1996.
Lawrence PK, Nelson WR, Liu W, Knowles DP, Foreyt WJ, Srikumaran S. b2 integrin Mac-1 is a receptor for Mannheimia haemolytica leukotoxin on bovine and ovine leukocytes. Vet Immunol Immunopathol 122: 285-294, 2008.
Lee CW, Shewen PE, Cladman WM, Conlon JA, Mellors A, Lo RY. Sialoglycoprotease of Pasteurella haemolytica A1: detection of antisialoglycoprotease antibodies in sera of calves. Can J Vet Res 2: 93-98, 1994.
Lee CW, Shewen PE. Evidence of bovine immunoglobulin G1 (IgG1) protease activity in partially purified culture supernate of Pasteurella haemolytica A1. Can J Vet Res 2: 127-132, 1996.
Lee RWH, Cornelisse M, Ziauddin A, Slack PJ, Hodgins DC, Strommer JN, Shewen PE, Lo RY. Expression of a modified Mannheimia haemolytica GS60 outer membrane lipoprotein in transgenic alfalfa for the development of an edible vaccine against bovine pneumonic pasteurellosis. J Biotechnol 2: 224-231, 2008.
Leite F, Gyles S, Atapattu D, Maheswaran SK, Czuprynski CJ. Prior exposure to Mannheimia haemolytica leukotoxin or LPS enhances β2-integrin expression by bovine neutrophils and augments LKT cytotoxicity. Microb Pathog 34: 267-275, 2003.
Lillic LE, Thomson RG. The pulmonary cleatance of bacteria by calves and mice. Can J Comp Med 36: 129-137, 1972.
Lo RYC, Mellors A. The isolation of recombinant plasmids expressing secreted antigens of Pasteurella haemolytica A1 and the characterization of an immunogenic 60 kDa antigen.Vet Microbiol 3: 381-391, 1996.
Lo RYC. Genetic analysis of virulence factors of Mannheimia (Pasteurella) haemolytica A1. Vet Microbiol 83: 23-35, 2001.
Lo RYC, Sathiamoorthy S, Shewen PE. Analysis of in vivo expressed genes in Mannheimia haemolytica A1. FEMS Microbiol Lett 1: 18-25, 2006.
Lopez A, Thomson RG, Savan M. The pulmonary clearance of Pasteurella haemolytica in calves infected with bovine parainfluenza 3 virus. Can J Comp Med 40: 385-391, 1976.
Madan Babu M, Sankaran K. DOLOP- database of bacterial lipoproteins. Bioinformatics 18: 641-643, 2002.
McVicker JK, Tabatabai LB. Isolation of immunogenic outer membrane proteins from Mannheimia haemolytica serotype 1 by use of selective extraction and immunoaffinity chromatography. Am J Vet Res 12: 1634-1640, 2002.
Meza LJ, Romero FA , G¨uemes FS, Tavera FT. Challenge exposure of sheep immunized with live vaccine and culture supernatant of Mannheimia haemolytica A1: Effects of revaccination. Small Rumin Res 70: 209-217, 2007.
Morck DW, Olson ME, Acres SD, Daoust PY, Costerton JW. Presence of bacterial glycocalyx and fimbriae on Pasteurella haemolytica in feedlot cattle with pneumonic pasteurellosis. Can J Vet Res 2: 167-171, 1989.
Morton RJ, Panciera RJ, Fulton RW, Frank GH, Ewing SA, Homer JT, Confer AW. Vaccination of cattle with outer membrane protein-enriched gractions of Pasteurella haemolytica and resistance against experimental challenge. Am J Vet Res 56: 875-879, 1995.
Morton RJ, Simons KR, Confer AW. Major outer membrane proteins of Pasteurella haemolytica serovars 1-15: comparisonof separation techniques and surfaceexposed proteins on selected serovars. Vet Microbiol 51: 319-330, 1996.
Mosier DA, Confer AW, Panciera RJ. The evolution of vaccines for bovine pneumonic pasteurellosis. Res Vet Sci 1: 1-10, 1989.
Mosier DA. Bacterial pneumonia. Bov Resp DisUpdate 13: 483-493, 1997.
Narayanan SK, Nagaraja TG, Chengappa MM, Stewart GC. Leukotoxins of gram-negative bacteria. Vet Microbiol 84: 337-356, 2002.
Newson IE, Cross F. Some bipolar organisms found in pneumonia in sheep. J Am Vet Med. Assoc 80: 711-719, 1932.
Noti JD, Johnson AK, Dillon JD. Structural and functional characterization of the leukocyte integrin gene CD11d. J Biol Chem 275: 8959-8969, 2000.
O’Brien JK, Duffus WP. Pasteurella haemolytica cytotoxin: relative susceptibility of bovine leucocytes. Vet Microbiol 13: 321-334, 1987.
Pandher K, Murphy GL. Genetic and immunological analyses of a 38 kDa surface-exposed lipoprotein of Pasteurella haemolytica A1. Vet Microbiol 51: 331-341, 1996.
Pandher K, Confer AW, Murphy GL. Genetic and immunologic analyses of PlpE, a lipoprotein important in complement-mediated killing of Pasteurella haemolytica serotype 1. Infect Immun 66: 5613-5619, 1998.
Pandher K, Murphy GL, Confer AW. Identification of immunogenic, surface-exposed outer membrane proteins of Pasteurella haemolytica serotype 1. Vet Microbiol 65: 215-226, 1999.
Potter MD, Lo RYC. Cloning and characterization of a gene from Pasteurella haemolytica A1 involved in lipopolysaccharide biosynthesis. FEMS Microbiol Lett 129: 75-82, 1995.
Purdy CW, Richards AB, Foster GS. Market stressassociated changes in serum complement activity in feeder calves. Am J Vet Res 52: 1842-1847, 1991.
Qrinn PJ. Clinical Veterinary Microbiology. London: Wolfe, pp254-258.
Rice JA, Carrasco-Medina L, Hodgins DC, Shewen PE. Mannheimia haemolytica and bovine respiratory disease. Anim Health Res Rev 2: 117-28, 2008.
Ribble CS, Meek AH, Shewen PE, Guichon PT, Jim GK. Effect of pretransit mixing on fatal fibrinous pneumonia in calves. J Am Vet Med Assoc 207: 616-619, 1995.
Rossi C, Kiesel GK. Susecptibility of bovine macrophages and tracheal-ring cultures to bovine viruses. Am J Vet Res 38: 1705-1708, 1997.
Sabri MY, Zamri-Saad M, Mutalib AR, Israf DA, Muniandy N. Efficacy of an outer membrane protein of Pasteurella haemolytica A2, A7 or A9-enriched vaccine against intratracheal challenge exposure in sheep. Vet Microbiol 73: 13-23, 2000.
Schaller A, Kuhnert P, Puente-Redondo VA, Nicolet J, Frey J. Apx toxins in Pasteurellaceae species from animals. Vet Microbiol 74: 365-376, 2000.
Shewen PE, Carrasco-Medina L, McBey BA, Hodgins DC. Challenges in mucosal vaccination of cattle. Vet Immunol Immunopathol 15: 192-198, 2009.
Slocombe RF, Seerksen Fj, Robinson NE, Trapp A, Gupta A, Newman JP. Interactions of cold stress and Pasteurella haemolytica in the pathogenesis of pneumonic pasteurellosis in calves: method of induction and hemotologic and pathologic changes. Am J Vet Res 45: 1757-1763, 1984.
Slocombe RF, Malark J, Ingersoll R, Derksen FJ, Robinson NE. Importance of neutrophils in the pathogenesis of acute pneumonic pasteurellosis in calves. Am J Vet Res 46: 2253-2258, 1985.
Smith GR. The characteristics of two types of Pasteurella haemolytica associated with different pathological conditions of sheep. J Pathol Bacteriol 81: 431-440, 1961.
Speer NC, Young C, Roeber D. The importance of preventing bovine respiratory disease: a beef industry review. Bovine Pract 35: 189-196, 2001.
Stevens PK, Czuprynski CJ. Pasteurella haemolytica leukotoxin induces bovine leukocytes to undergo morphologic changes consistent with apoptosis in vitro. Infect Immun 64: 2687-2694, 1996
Stolzenberg ED, Anderson GM, Ackermann MR, Whitlock RH, Zasloff M. Epithelial antibiotic induced in states of disease. Proc Natl Acad Sci USA 94: 8686-8690, 1997.
Straus DC, Purdy CW, Loan RW, Briggs RF, Frank GH. In vivo production of neuraminidase by Pasteurella haemolytica in market stressed cattle after natural infection. Curr Microbiol 4: 240-244, 1998.
Tato CM, Hunter CA. Host-pathogen interactions: subversion and utilization of the NF-kappa B pathway during infection. Infect Immun 70: 3311-3317, 2002.
Thumbikat P, Briggs RE, Kannan MS, Maheswaran SK. Biological effects of two genetically defined leukotoxin mutants of Mannheimia haemolytica. Microb Pathog 34: 217-226, 2003.
Tian L, Yoshihara Y, Mizuno T, Mori K, Gahmberg CG. The neuronal glycoprotein telencephalin is a cellular ligand for the CD11a/CD18 leukocyte integrin. J Immunol 158: 928-936, 1997.
Walker RD, Hopkins FM, Schultz TW, McCracken MD, Moore RN. Changes in leukocyte populations in pulmonary lavage fluids of calves after inhalation of Pasteurella haemolytica. Am J Vet Res 46: 2429-2433, 1985.
Welch R.A. RTX toxin structure and function: a story of numerous anomalies and few analogies in toxin biology. Curr Top Microbiol Immunol 257: 85-111, 2001.
Weldon SK, Mosier DA, Simons KR, Craven RC, Confer AW. Identification of a potentially important antigen of Pasteurella haemolytica.Vet Microbiol 3-4: 283-291, 1994.
Welsh RD, Dye LB, Payton ME, Confer AW. Isolation and antimicrobial susceptibilities of bacterial pathogens from bovine pneumonia: 1994-2002. J Vet Diagn Invest 5: 426-431, 2004.
Whiteley LO, Maheswaran SK, Weiss DJ, Ames TR, Kannan MS. Pasteurella haemolytica A1 and bovine respiratory disease: pathogenesis.J Vet Intern Med 1: 11-22, 1992.
Woldehiwet Z, Sharma R. Evidence of immunosuppression by bovine respiratory syncytial virus. Scand J Immunol 11: 75-80, 1992.
Yates WD. A review of infectious bovine rhinotracheitis, shipping fever pneumonia and viral-bacterial synergism in respiratory disease of cattle. Can J Comp Med 46: 225-263, 1982.
Yin Y, Zhang F, Ling V, Arrowsmith CH. Structural analysis and comparison of the C-terminal transport signal domains of hemolysin A and leukotoxin A. FEBS Lett 366: 1-5, 1995.
Yoo HS, Rajagopal BS, Maheswaran SK, Ames TR. Purified Pasteurella haemolytica leukotoxin induces expression of inflammatory cytokines from bovine alveolar macrophages. Microb Pathog 18: 237-252, 1995.
Younan M, Fodor L. Characterization of a new Pasteurella haemolytica serotype (A17). Res Vet Sci 58: 98, 1995.
Zecchinon L . How Mannheimia haemolytica defeats host defence through a kiss of death mechanism. Vet Res 36: 133-156, 2005.