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1.
通过不同攻毒途径、不同攻毒剂量、不同采血时间及不同采血途径的试验表明，以翅静脉途径攻击多杀巴氏杆菌，剂量10亿／只，在鸡濒死挣扎时的即刻颈静脉无菌放血，可以自鸡体采集到最大量的含菌血，每只鸡37～39mL，再将血液进行差速和蔗糖密度梯度离心，可最大限度地提取到血液中的巴氏杆菌，每只鸡300～400亿的总菌量。该结果与报道的提取方法相比，效率提高了1000倍以上。提取的细菌灭活后免疫鸡，经异型菌交叉攻毒试验表明，体内繁殖的巴氏杆菌具有交叉保护特性，而体外培养菌则没有。用该方法提取的巴氏杆菌，其总蛋白和总糖含量均高于培养基培养菌，提示二者之间的差异可能与交叉保护特性存在着一定的相关性。 相似文献
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禽多杀巴氏杆菌交叉保护因子的研究 总被引：7，自引：0，他引：7
禽多杀巴氏杆菌交叉保护因子存在于鸡活体培养物中，用鸡活体培养禽多杀巴氏杆菌Ｃ４８－１强毒株（Ｉ型），菌血症时收获细菌制成死菌苗，免疫鸡对异型菌株Ｐ１０５９（３型）攻击产生交叉保护，而４１℃肉汤培养时Ｃ４８－１菌不具备这种交叉保护，活体培养的细菌冻融后对异型菌株保护率达１００％，经溶菌酶等处理溶解后的溶解物上清及沉淀对异型菌株产生６６、７％和８０％的交叉保护，但其沉淀经蛋白酶处理后则失去了交叉保护能 相似文献
4.
选择禽多杀性巴氏杆菌Ｃ４８－１强毒株（血清１型），接种鸡胚，收集死胚制备组织灭活油乳苗，分别免疫鸡和鸭，设肉汤培养的Ｃ４８－１菌株灭活乳化苗作免疫对照。免疫后４周用异源血清型禽多杀性巴氏杆菌Ｐ１０５９（血清３型）攻击。经组织灭活油乳苗免疫的鸡保护率达９３．３％，鸭保护率达１００％，而经肉汤培养的灭活苗对鸡和鸭均无保护作用，说明鸡胚培养的多杀性巴氏杆菌具有交叉保护作用。 相似文献
5.
禽霍乱鸡胚组织灭活油乳苗的研究 总被引：2，自引：0，他引：2
选择禽多杀性巴杆菌Ｃ４８－１强毒株（血清１型）接种鸡胚，收集死胚制备组织灭活油乳苗，分别免疫鸡和鸭，设肉汤培养的Ｃ４８－１菌株灭活乳化苗免疫对照，免疫后４周用异源血清型禽多杀性巴氏杆菌Ｐ１０５９（血清３型）攻击。经组织灭活油乳免疫的鸡保护率达９３．３％，鸭保护率达１００％，而经肉汤培养的灭活苗对鸡和鸭均无保护作用，说明鸡胚培养的多杀性巴氏杆菌具有交叉保护作用。 相似文献
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将鸡多杀性巴氏杆菌菌液、大肠杆菌菌液分别经甲醛灭活,加氢氧化铝胶,弃去部分上清液,将二者按3:1的比例混合制成二联灭活疫苗.疫苗接种鸡安全;接种21 d后对免疫鸡攻毒,免疫鸡对两种疫病的免疫保护均达到3/4以上,说明将鸡巴氏杆菌、大肠杆菌制成二联灭活疫苗是完全可行的. 相似文献
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禽多杀巴氏杆菌交叉保护因子（ＣＰＦ）存在于鸡活体培养物中．用鸡活体培养禽多杀巴氏杆菌Ｃ４８－１强毒株（Ⅰ型），菌血症时收获细菌制成死菌苗，免疫鸡对异型菌株Ｐ１０５９（３型）攻击产生交叉保护，而４１℃肉汤培养时Ｃ４８－１菌株不具备这种交叉保护．活体培养的细菌冻融后对异型菌株保护率达１００％，经溶菌酶等处理溶解后的溶解物上清及沉淀对异型菌株产生６６．７％和８０％的交叉保护．但其沉淀经蛋白酶处理后则失去了交叉保护能力．生化分析表明，活体培养的细菌与肉汤培养的细菌其蛋白质与糖含量比不同；ＳＤＳ－ＰＡＧＥ分析表明，有一分子量约１９３２０蛋白成分仅存在于活体培养的细菌中． 相似文献
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在猪丹毒、多杀性巴氏杆菌病二联灭活疫苗的效力检验中,需要进行动物攻毒保护试验。传统方法使用的是2~8 ℃保存的新鲜菌液对试验动物进行攻毒。本研究中使用的是低温保存的冻干菌液,并对冻干保存的菌数进行了测定和优化。结果表明,冻干保存的菌液能达到新鲜菌液同等的检验结果。同时,对冻干菌液保存期进行试验验证,发现细菌存活状态稳定。因此,在猪丹毒、多杀性巴氏杆菌病二联灭活疫苗效力检验中,可将新鲜的攻毒菌液冻干保存,用于攻毒保护试验,以避免每次繁重的新鲜菌液前期准备工作,并克服活菌数衰减所造成的检验结果不准确等问题。 相似文献
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用５批禽巴氏杆菌Ｂ２６－Ｔ１２００弱毒疫苗共接种３００万只鸡．现场观察６２４３７只免疫鸡．结果表明，该菌对不同品种鸡安全．只有３．８％免鸡表现为一过性食欲减少、精神稍差．但２天内都能恢复正常，没有引起免疫鸡直接死亡现象．田间免疫１４０天抽取各批苗免疫鸡攻毒，有８０％（１６／２０）鸡得到保护，免疫鸡群在免疫１４０天内没有发生禽巴氏杯菌病． 相似文献
10.
用５批禽巴氏杆菌Ｂ２６－Ｔ１２００弱毒疫苗共接种３００万只鸡。现场观察６２４３７只免疫鸡。结果表明，该苗对不同品种鸡安全，只有３．８％免鸡表现为一过性食欲减少，精神稍差，但２天内都能恢复正常，没有引起免疫鸡直接死亡现象。田间免疫１４０天抽取各批苗免疫鸡攻毒，有８０％（１６／２０）鸡得到保护，免疫鸡群在免疫１４０天内没有发生禽巴氏杆菌病。 相似文献
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A 39 kDa protein mediates adhesion of avian Pasteurella multocida to chicken embryo fibroblast cells 总被引：5，自引：0，他引：5
Borrathybay E Sawada T Kataoka Y Ohtsu N Takagi M Nakamura S Kawamoto E 《Veterinary microbiology》2003,97(3-4):229-243
To clarify the role of avian Pasteurella multocida capsule in pathogenesis, adhesion of capsulated strains P-1059, X-73 and Pm-18, and noncapsulated strains P-1059B, Pm-1 and Pm-3 to chicken embryo fibroblast (CEF) cells was compared. Number of adherent organisms of the capsulated strains to CEF cells were approximately three times as much as noncapsulated strains indicating that adhesive properties were enhanced by the presence of bacterial capsule. Pretreatments of the bacterial cells with heat, trypsin, or with antiserum caused a marked decrease in adhesion of capsulated strain P-1059 and its noncapsulated variant P-1059B. However, depolymerization of capsular hyaluronic acid with high dose of hyaluronidase enhanced adhesion of these strains. Combined treatments of the bacterial cells with both hyaluronidase and trypsin significantly (P < 0.05) inhibited the adherence of strain P-1059 as compared to the treatment only with trypsin, but strain P-1059B was not affected. SDS-PAGE profiles of crude capsular extract (CCE) prepared from capsulated strain P-1059 and its noncapsulated variant P-1059B grown on dextrose starch agar (DSA) plates by heating at 56 degrees C in a 2.5% NaCl solution demonstrated eight protein bands of 28, 34, 36, 39, 52, 56, 63 and 93 kDa. The 28, 34 and 36 kDa proteins were commonly major for both strains, and the 39 kDa protein was major only for strain P-1059 but poor in strain P-1059B. Outer membrane protein (OMP) profiles were identical with a major protein at 34 kDa and four minor proteins between the two strains. The adhesion of strain P-1059 and strain P-1059B to CEF cells was inhibited significantly (P < 0.01) by treatment with rabbit antisera against P-1059, P-1059B, CCE or 39 kDa protein of strain P-1059 as compared to the treatment with either PBS or with normal rabbit serum. These results indicated that an antigenic 39 kDa protein in the capsule may be responsible for adhesion of avian P. multocida type A strains to CEF cells as a virulence factor. 相似文献
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Association of complement sensitivity with virulence of Pasteurella multocida isolated from turkeys 总被引：1，自引：0，他引：1
Two strains of Pasteurella multocida, both derivatives of strain P1059, were compared for virulence for 14-week-old turkeys and sensitivity to turkey plasma. Strain P1059-1, a nalidixic-acid-resistant mutant of P1059 with an LD50 of approximately 10(3) colony-forming units (CFU), was more resistant to the bactericidal effects of fresh turkey plasma at 37 C than avirulent strain P1059-1A. P1059-1A, with an LD50 of approximately 10(8) CFU, is an acapsular variant of P1059-1 that spontaneously arose after prolonged passage on artificial medium. The bactericidal effect on P1059-1A was removed when turkey plasma was treated with heat or with zymosan, maneuvers that removed hemolytic complement activity from turkey plasma. 相似文献
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The avian strain P-1059 of Pasteurella multocida was grown on blood agar (BA), on dextrose-starch agar (DSA), or in Heddleston's hydrogen sulfide test broth. Cells were examined for the presence of pili using electron microscopy after staining with phosphotungstic acid, and they were examined for capsule after ruthenium red staining. Pili were found on the capsulated iridescent type, P-1059I, and on two non-capsulated variants, the blue, P-1059B, and the gray, P-1059G. Many cells grown on BA were heavily piliated. In contrast, fewer cells grown on DSA had pili, and piliation was only slight to moderate. The P-1059I, P-1059B, and P-1059G produced pellicles when grown on broth medium. Pili were found on the circumference of the cells grown on either agar or broth medium. Occasionally a pilus connecting two cells was seen on cells cultured in broth. Cultivation of the P-1059I on DSA containing the iron-chelating agent alpha,alpha'-bipyridyl produced a non-capsulated blue variant. The non-capsulated variant reverted to P-1059I when grown on BA but did not revert when grown on DSA. 相似文献
14.
Borrathybay E Sawada T Kataoka Y Okiyama E Kawamoto E Amao H 《Veterinary microbiology》2003,97(3-4):215-227
Capsule thickness of avian Pasteurella multocida type A strains was determined by transmission electron microscopy after labeling with polycationic ferritin and compared with their pathogenicity for chickens. The capsule thickness of P. multocida strains Pm-18 and X-73 was 81.4 and 50.1 nm on average, respectively. These strains were highly virulent for chicken, whereas the less virulent strains Pm-1 and Pm-3 had a thin and irregular capsule, 21.0 and 29.8 nm on average, respectively. However, the thickest capsule was observed in strain P-1059, 101.2 nm on average, and the strain revealed moderate virulence. The noncapsulated variant P-1059B, which was derived from strain P-1059, revealed low virulence. The six P. multocida strains were examined with regard to protein content on the capsule of organisms. Amounts of total proteins of crude capsular extract (CCE) from capsulated strains were approximately twice those of the noncapsulated strains. The amount of an antigenic 39 kDa protein in the CCE were found to correlate with the capsule thickness, since heavily capsulated strains exhibited the greatest amount, whereas noncapsulated strains including noncapsulated and low virulent variant P-1059B possessed little 39 kDa protein. The results demonstrated that the capsule thickness and the quantity of a 39 kDa capsular protein of avian P. multocida type A strains correlated with their pathogenicity for chickens. 相似文献
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Encapsulated avian strains of Pasteurella multocida possessing an A-type capsule were shown to be resistant to the bactericidal action of turkey serum, whereas unencapsulated variants as well as other unencapsulated strains were not. Removal of the capsule from serum-resistant strain P1059-1 resulted in this strain becoming susceptible to the bactericidal effects of turkey serum. Since complement was consumed when encapsulated or unencapsulated strain P1059-1 was incubated in turkey serum, we conclude that the capsule acts to shield the outer membrane rather than prohibiting the generation of an effective membrane attack complex. 相似文献
16.
The role of a 39kDa protein of avian Pasteurella multocida in pathogenesis of fowl cholera was investigated using monoclonal antibodies (Mabs). Mabs were prepared by immunization of BALB/c mice with a crude capsular extract (CCE) of P. multocida strain P-1059 (serovar A:3). Totally eight hybridomas producing Mab were obtained. Immunoblot analysis of the hybridomas revealed that all the Mabs recognized a 39kDa protein of CCE. Treatment of CCE antigen with proteinase K or periodic acid indicated that the epitope recognized was proteinaceous. The Mabs reacted with a major 39kDa protein of CCE from encapsulated strains but not with any protein of non-capsulated strains indicating that a direct correlation between encapsulation and the 39kDa protein. Immunoelectron microscopy on strain P-1059 and the non-capsulated derivative P-1059B (serovar -:3) reacting with the Mabs and gold-labeled anti-mouse IgG indicated that the protein is associated with the capsule. The Mabs significantly inhibited the adherence of encapsulated P. multocida strains to chicken embryo fibroblast cells, but only slightly that of non-capsulated strains. Mice passively immunized with the Mabs were protected from lethal challenge with virulent strains P-1059 and X-73 (serovar A:1). Thus the capsular 39kDa protein was determined to be an adherence factor and a cross-protective antigen of avian P. multocida type A strains. 相似文献
17.
Characterization of outer membrane protein-enriched extracts from Pasteurella multocida isolated from turkeys 总被引：1，自引：0，他引：1
Outer membrane protein (OMP)-enriched extracts of avian strains of Pasteurella multocida were examined by use of sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Culture medium did not have a significant effect on the OMP profiles of strains of P multocida examined; however, in vivo propagation had an appreciable effect on the OMP profile composition of the reference strain P-1059. Such bacteria, expressed several additional OMP in the 27-kD, 48-kD, 56-kD, 60-kD, 80-kD, and 94-kD molecular mass regions. These OMP were not detected in the electrophorogram of strain P-1059 grown in vitro. The OMP profiles of reference strains of the 16 serotypes of P multocida did not identify any serotype-specific protein markers. Field strains of serotype A:3 had variation in OMP profiles and did not express OMP that all were identical to that expressed by the reference strain P-1059. The live attenuated CU and M9 bacterial vaccine strains expressed strain-specific OMP markers of 48-kD and 45-kD molecular masses, respectively. These strain-specific OMP markers may be used to differentiate these strains from virulent field strains that are of the same serotype and isolated from turkeys that have succumbed to pasteurellosis as a result of vaccine-related reactions or breakdown in immunity. 相似文献
18.
Ali HA Sawada T Noda K 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2004,66(12):1603-1604
A 39 kDa protein of avian Pasteurella multocida strain P-1059 (serovar A:3) was purified from a crude capsular extract by immunoaffinity chromatography by using a ligand of purified mouse monoclonal antibody to the 39 kDa capsular protein of the strain. Protective activity of the purified 39 kDa protein antigen was determined by inoculation of ddY mice twice with 25 or 125 mug of the protein and challenge-exposure with 10 or 50 LD(50) of strains P-1059 or X-73 (serovar A:1). The results showed that the antigen gave high protection (60 to 100%). These results indicated that the 39 kDa protein of avian P. multocida is a cross-protective antigen over serovars A:1 and A:3. 相似文献
19.
The role of the capsule in the pathogenesis of fowl cholera was studied in turkeys. Avian Pasteurella multocida P-1059 was used in an encapsulated form, an enzymatically decapsulated form, and a mutant form lacking capsule-productivity (strain T-325). These forms were inoculated intravenously into normal or immune turkeys, and the numbers of viable bacteria in the blood, liver, and spleen were enumerated over a 120-minute period. Both normal and immune birds rapidly removed all three forms of organisms from the blood-stream at similar rates and trapped in the liver and spleen. In the liver of normal birds, the non-encapsulated mutant T-325 was readily inactivated, but the encapsulated P-1059 strain was not. When the decapsulated form of P-1059 was used, the bacterial counts in the liver temporarily decreased at 60 minutes PI. In immune birds, all three forms of organisms were equally inactivated in the liver. In the spleen, however, the bacterial numbers did not change throughout 120 minutes PI with all three forms of organisms in both normal and immune turkeys. The results indicated that the blood-borne P. multocida were readily trapped by reticuloendothelial phagocytes. The trapping process was not affected by encapsulation of the organism or by the immune status of turkey. Both factors, however, appeared to greatly influence the subsequent killing of P. multocida by hepatic, but not splenic, phagocytes. 相似文献
20.
Sthitmatee N Kataoka Y Sawada T 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2011,73(11):1445-1451
A mutant strain, PBA322, was constructed by electroporation of a phagemid containing the coding region of antisense RNA of the ompH gene, encoding 39 kDa capsular protein or OmpH, into the parental strain P-1059 (serovar A:3) of Pasteurella multocida, and the pathogenicity was determined in mice and chickens. Grayish colonies of the mutant, indicating loss of capsule synthesis, were observed under a stereomicroscope using obliquely transmitted light, while iridescent colonies were observed for the parental strain. Moreover, strain PBA322 showed a low amount of OmpH compared with the parental strain on SDS-PAGE. Additionally, the capsule of strain PBA322 was thinner than that of the parental strain according to electron microscopy, correlating to the attenuation against chickens. In conclusion, strain PBA322, the mutant of P. multocida strain P-1059, was completely attenuated for chickens. 相似文献