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1.
Fluorescent antibody technique has been used as a diagnostic tool for the identification of pathogenic avian Mycoplasma species. Fluorescein-conjugated antisera for Mycoplasma iowae prepared from rabbit hyperimmune serum against one serotype (I) has not always been reliable, giving false-negative results. A fluorescein-conjugated antisera was prepared with antibody against the six identified serotypes (I, J, K, N, Q, and R) that comprise the species M. iowae. This heterogeneous conjugate was used to positively identify M. iowae in field isolates and lab strains, while maintaining specificity. Polymerase chain reaction specific for M. iowae was used to determine specificity. 相似文献
2.
A dot-immunobinding assay, amplified with avidin and biotin (DAB assay), was used to detect serum antibodies to Mycoplasma iowae in immunized turkeys. The DAB assay was used to test serum samples from 122 commercial market turkey flocks obtained from four Iowa processing plants. The samples were pooled and tested for the presence of antibodies to four species of Mycoplasma spp. considered to be important pathogens for turkeys: M. gallisepticum (MG), M. iowae (MI), M. meleagridis (MM), and M. synoviae (MS). The occurrence of antibodies against these mycoplasmas, as determined by the DAB assay, were 5.7% for MG, 18.0% for MI, 77.9% for MM, and 9.8% for MS. 相似文献
3.
传染性法氏囊病病毒广西分离株血清亚型的确定 总被引：2，自引：0，他引：2
应用兔制备的抗传染性法氏囊病病毒(IBDV)高免血清,在鸡胚成纤维细胞(CEF)上对从广西发病鸡群分离的4个IBDV代表性流行毒株和2个常用疫苗株进行交叉中和试验。结果表明6个毒株被分为2个血清亚型;根据试验所得的R值,应用聚类分析法分析了各亚型毒株之间的亲缘关系,结果显示目前在广西流行的IBDV野毒株之间以及其与疫苗株间的抗原性存在一定的差异。研究结果对及时掌握广西IBDV流行毒株的抗原变异并为研制更有效的适合本地使用的IBD疫苗提供了科学依据。 相似文献
4.
Western blots of proteins of 14 Mycoplasma iowae strains and isolates resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis were probed with three monoclonal antibodies (MAbs), MI6, MI7, and MI8. MAb MI6 reacted with one or more antigens with apparent molecular weights of 60,000, 70,000, and 94,000. In three strains (N-PHN-D13, R-D2497, and K 1805), antigens located on a single peptide band were recognized, while in others additional epitopes at different molecular-weight positions were revealed. A similar pattern was observed with MAb MI7, although it reacted with fewer antigens than did MAb MI6 and failed to recognize antigens in strains N-PHN-D13 and R-D2497. MAb MI8 reacted with an antigen at an apparent molecular-weight position of 28,000 in four of the 14 strains and isolates. The diverse reaction patterns observed with the MAbs in the 14 M. iowae strains and isolates confirms the occurrence of antigenic variation within this species. Antigenic variation in M. iowae may be pivotal in determining host-parasite interactions, pathogenesis, and the outcome of disease. 相似文献
5.
Immunoblotting was used to check the antigenic profiling of 27 Mycoplasma meleagridis strains isolated in different countries. Hyperimmune polyclonal rabbit antiserum as well as monoclonal antibodies (MAbs) raised against M. meleagridis (MM) showed antigen heterogeneity among strains. Five anti-MM MAbs were selected for lack of reaction against heterologous avian mycoplasma. Three of these five Mabs did not cross-react with 63 mycoplasma strains from six species affecting turkeys other than M. meleagridis. The five Mabs used to analyse the epitopes of 30 M. meleagridis strains indicated that some epitopes were not expressed in all strains. Moreover, other epitopes were located on proteins which differed according to number or molecular mass from strain to strain. The five Mabs therefore, recognised variable surface proteins, among which two were amphiphilic membrane proteins.Three of the selected Mabs recognised 29 or 30 of the 30 tested strains. The in vitro expression of surface epitopes in M. meleagridis ATCC 25284 was investigated by colony immunobinding and allowed demonstration of a variable antigenic system. 相似文献
6.
Six field strains of Mycoplasma hyopneumoniae isolated from pneumonic lungs of pigs, reference strains 11 and J of M hyopneumoniae, Ms 42 strain of Mycoplasma flocculare, and BTS 7 strain of Mycoplasma hyorhinis were compared serologically, using hyperimmune antisera produced in rabbits. All strains of M hyopneumoniae were closely related as determined with the disk growth-inhibition test; however, differences in zone sizes indicated that some antigenic heterogeneity existed. Cross-reactions were not detected between M hyopneumoniae, M flocculare, and M hyorhinis with the growth-inhibition test. The metabolic-inhibition test was more useful for detection of intraspecies antigenic difference than was the growth-inhibition test, since antigenic diversity was clearly detected among M hyopneumoniae strains. Slight cross-reactions were observed between M hyopneumoniae and M flocculare. Using 2-dimensional immunoelectrophoresis, antigenic differences were observed among M hyopneumoniae strains, although many common components also were detected in electropherograms. Mycoplasma flocculare possessed a close antigenic relationship to M hyopneumoniae, as determined by two-dimensional immunoelectrophoresis, whereas both organisms were less related to M hyorhinis. Evidence obtained in this study indicated that strains of mycoplasmas tentatively identified as M hyopneumoniae were similar antigenically, but evidence was obtained also of some diversity in antigenic structure among these strains. 相似文献
7.
Bercic RL Slavec B Lavric M Narat M Zorman-Rojs O Dovc P Bencina D 《Veterinary microbiology》2008,130(3-4):391-397
Among 23 currently recognized avian Mycoplasma (AM) species only Mycoplasma gallisepticum, Mycoplasma synoviae, Mycoplasma meleagridis and Mycoplasma iowae cause disease and loss of production in chickens and/or turkeys. Because neuraminidases are considered virulence factors in many pathogenic microorganisms the aim of our study was to determine which AM species possess neuraminidase enzymatic activity (NEAC). Small samples of AM cells were assayed for NEAC using the chromogenic substrate 5-bromo-4-chloro-3-indolyl-alpha-d-N-acetylneuraminic acid. In the case of positive NEAC reaction the substrate gave the insoluble indigoblue product what enabled simple test and easy estimation of NEAC. M. gallisepticum and M. synoviae which share sequences of the gene encoding neuraminidase (sialidase NanH) exhibited considerable levels of NEAC. However, NEAC levels differed among their strains, as well as among cultures of different strains. Only certain cultures of the type strain of M. meleagridis showed NEAC, whereas among six serovars of M. iowae only serovar I (type strain 695) showed NEAC. Weak NEAC was detectable in M. anseris, M. cloacale and M. pullorum, whereas the type strain of M. corogypsi (BV1) showed strong NEAC. Our study provides novel informations about NEAC in AM species and suggests that higher invasiveness and possibly, the pathological processes might be associated with their NEAC. 相似文献
8.
Three-week-old turkeys were inoculated intranasally with approximately 10(6) colony-forming units (CFU) of putative variant Mycoplasma gallisepticum (MG) strains M876, M35, or the virulent S6 reference strain. Uninoculated turkeys in each group served as contact sentinels. The hemagglutination-inhibition (HI) test and enzyme-linked immunosorbent assay (ELISA) were used to determine serologic responses. MG was isolated from 100% and 92% of S6- and M876-inoculated turkeys, respectively, on day 7 PI. However, culture-positive rates among M876-inoculated turkeys declined more rapidly, transmission to contact sentinels took longer and occurred at lower rates, and serologic responses measured by HI and ELISA were lower than in S6-infected turkeys. Testing sera from inoculated turkeys for antibodies to MG in homologous and heterologous ELISA systems indicated that strain M876 was significantly (P less than 0.05) less immunogenic than S6 (days 62 and 95 PI), and that the homologous ELISA was more sensitive (P less than 0.005). MG strain M35 failed to infect turkeys in three attempts, even though the inocula used were viable on culture media. 相似文献
9.
应用病毒感染的鸡胚材料免疫新西兰兔的方法制备抗鸡传染性支气管炎病毒(IBV)单因子血清,然后在鸡胚气管环培养(Tracheal organ cultures,TOC)上对广西分离的7个IBV代表性毒株和3个常用疫苗株进行交叉病毒中和试验。结果显示,10个毒株被分为6个血清型。根据试验所得的R值,应用聚类分析法分析了各血清型毒株之间的亲缘关系,显示目前在广西流行的IBV野毒株之间以及其与疫苗株间的抗原性存在很大程度的差异,分属不同的血清型。同时还对IBV基因分型和血清分型之间的关系进行了探讨。 相似文献
10.
Two putative variant Mycoplasma gallisepticum (MG) strains (M876 and M35), originally isolated from commercial turkeys, were compared with eight well-characterized MG strains by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). SDS-PAGE protein profiles indicated that the variant strains were correctly classified as MG based on homologous patterns in species-specific regions of the electrophoretic profiles. However, differences in protein profiles also indicated that variant strains M876 and M35 were different from each other and the other MG strains tested. Immunoblotting was used to assess the humoral immune response of turkeys to infection with the S6 reference strain or M876 variant strain of MG. Immunoblots using antisera to M876 showed that seroconversion to this isolate was slower, and to fewer MG proteins when compared with immunoblots using antisera to S6. Immunoblot analyses further indicated that pooled antisera from turkeys inoculated with either S6 or M876 reacted with each of 10 MG strains tested. However, pooled S6 antisera reacted with greater intensity and with more MG proteins than did pooled M876 antisera. The species-specific immunodominant proteins with the greatest potential for use as antigens in serologic tests appeared to be those of 64 (p64) and 56 (p56) kilodaltons molecular mass. 相似文献
11.
K R Rhoades 《Avian diseases》1975,19(3):437-442
The antibody response of turkeys exposed to Mycoplasma synoviae intravenously and by way of air sacs was determined by tube agglutination, plate agglutination, hemagglutination-inhibition (HI), and gel diffusion precipitin tests. The results suggested that continued antigenic stimulation was lacking in most turkeys and that the response was due mostly to IgM-type immunoglobulin. Under those conditions, both types of agglutination tests were effective and were more sensitive indicators of exposure than the HI test. The gel diffusion precipitin test was not considered effective under the conditions of this study. HI activity occurred in serums of intravenously exposed turkeys within 4 days of exposure. The sensitivity of this activity to 2-mercaptoethanol treatment suggested that IgM was responsible. 相似文献
12.
All nine Mycoplasma iowae strains and one strain of M gallinarum grew on 0.05 per cent 'bile' agar medium. The colony size of M iowae on this agar medium was similar to the size obtained on bile-free mycoplasma agar. One strain each of M maculosum, M arginini and M bovis also grew on 0.05 per cent bile agar. However, one strain each of M gallisepticum and M meleagridis were inhibited at this concentration. Six of the nine strains of M iowae were also resistant to 1 per cent 'bile' in broth medium but all were resistant to 0.5 per cent. The resistance of M iowae to 0.5 per cent 'bile' in broth may be a useful characteristic for differentiating it from some of the other avian mycoplasmas. 相似文献
13.
14.
Strains of Mycoplasma iowae were homogeneous in some characteristics and heterogeneous in others. Thus, the biochemical tests, immunofluorescence, and protein profiling by sodium dodecyl sulfate-polyacrylamide gel electrophoresis were group-specific tests. However, some minor differences in protein patterns were seen among strains. The growth inhibition test tended to be strain-specific. Hemagglutination titers were very low and unstable for the majority of strains. One strain (RY-65) with a stable high-titer hemagglutinin failed to react in the hemagglutination-inhibition test against immune sera to the reference strains. Restriction endonuclease DNA analyses was the most useful method to differentiate 1 strain from another. 相似文献
15.
DNA fingerprinting for differentiation of field isolates from reference vaccine strains of Pasteurella multocida in turkeys 总被引：2，自引：0，他引：2
The genomes from field isolates of Pasteurella multocida in turkeys and those of P multocida reference CU and M9 vaccine strains were analyzed and compared after cleavage with restriction endonucleases. The electrophoretic profiles obtained with DNA fragments from field isolates and vaccine strains of the same serotype were characteristic and reproducible. These features indicated the existence of differences among the isolates of the same serotype that cannot currently be detected, using available serotyping methods. However, several field isolates had electrophoretic profiles similar to those of either CU or M9 vaccine strain. It was concluded that restriction endonuclease analysis of DNA genomes from P multocida isolated from turkeys provides the information for differentiation of field isolates from vaccine strains of the same serotype. 相似文献
16.
S M Elahi S Harpin E Cornaglia B Talbot Y Elazhary 《Canadian journal of veterinary research》1997,61(1):34-38
Antigenic variation among 13 Quebec isolates of bovine viral diarrhea virus (BVDV), 4 reference strains and 2 American isolates were studied by peroxidase-linked antibody assay (PLA assay) and neutralization test (NT). The Quebec strains consisted of 3 isolates before 1993 and 10 isolates from 1993. In the PLA assay, we compared 2 different fixatives, acetone and formalin. Acetone-fixation allowed us to identify 6 groups from amongst the viruses tested. All the Quebec isolates were different from the reference strains. In addition, antigenic variation was detected between Quebec isolates obtained before and during 1993. However, PLA assays performed after formalin fixation did not detect these antigenic variations. Neutralization tests were carried out with 2 polyclonal antibodies (PAb) and 6 monoclonal antibodies (MAb). They were used to classify BVDV strains and isolates into 4 groups and 7 subgroups respectively. In conclusion, we demonstrated that the BVDV isolates from the 1993 outbreak in Quebec are antigenically different from reference strains and from isolates existing in Quebec before 1993. In addition, we have shown that 2 internationally used fixation-methods in PLA assay give different results. The usefulness of each method is discussed. 相似文献
17.
Three antigens were prepared from Pasteurella multocida strain P-1059, and their immunogenicity and antigenic relationships were investigated. The 3 antigens were a soluble antigen purified from a 2.5% NaCl extract (2.5S), a similar antigen purified from an extract in 0.3% formalin solution containing 0.85% NaCl (FS), and lipopolysaccharide (LPS). The antigens were treated with various chemicals and enzymes to study their antigenic and immunogenic determinants. Antigenic analyses with ELISA inhibition tests indicated that 2.5S and FS were similar LPS-protein complex antigens. The 2.5S and FS antigens induced protective immunity in turkeys with high antibody titers against LPS antigen. Although LPS was a component of 2.5S and FS, LPS itself was poorly immunogenic in turkeys. The antigenicity of protein compounds in 2.5S was deteriorated by protease treatment, which, however, did not significantly diminish the protective immunogenicity. Treatment of 2.5S with sodium periodate, altering its carbohydrate moieties, decreased its immunogenicity. The immunogenicity of 2.5S also was abolished by phenol-water treatment, owing to dissociation of the LPS-protein complex. These findings suggest that a certain form of LPS-protein complex is essential for the induction of immunity against the P multocida infection in turkeys. 相似文献
18.
Hemorrhagic enteritis of turkeys: influence of maternal antibody and age at exposure 总被引：1，自引：0，他引：1
The effect of maternal antibody (MAB) to hemorrhagic enteritis (HE) on the response of turkeys to infection with virulent and avirulent strains of HE virus (HEV) was examined. The influence of age at exposure and treatment with HEV antibody on development of clinical HE also was studied. MAB protected poults from clinical HE for up to 6 weeks of age. MAB also interfered with vaccination against the disease for at least 5 weeks after hatching, as indicated by absence of HEV antigen in spleens and by poor seroconversion at 6 days and at 3 weeks post-vaccination, respectively. The incidence of clinical HE in MAB-negative poults was significantly higher in poults inoculated with virus at 15 days of age or older than in poults inoculated at 1-13 days of age. Further, MAB-negative poults embryonally inoculated with virulent or avirulent strains of HEV did not develop disease; these poults developed antibody and resisted challenge with virulent virus at 6 weeks of age. Poults treated with HE antibody within 1 hour of challenge or at 1, 3, or 5 weeks before challenge with virulent virus were protected against lesions and mortality induced by HEV. These results suggest that MAB may influence susceptibility of turkeys to infection with HEV for at least 5 to 6 weeks after hatching, unlike the case with most other viral infections of poultry. The results confirm that early age resistance to clinical HE is independent of MAB and suggest that such resistance persists for up to 13 days of age. The data also suggest that turkeys lacking MAB can be immunized against HE by embryo vaccination. 相似文献
19.
Isolation of a monoclonal antibody with specificity for commonly employed vaccine strains of Newcastle disease virus 总被引：2，自引：0，他引：2
A monoclonal antibody, AVS-I, was produced from a hybridization of murine myeloma cells and splenocytes from mice immunized with the La Sota strain of Newcastle disease virus (NDV). The hybridoma producing AVS-I, selected from 184 NDV-positive supernatants, is one of two supernatants that reacted exclusively with lentogenic strains in an indirect enzyme-linked immunosorbent assay. AVS-I can also be assayed by hemagglutination-inhibition (HI), which was used to test selected reference avian paramyxovirus (PMV) strains of types 1 to 3. NDV vaccines La Sota and B1 and field isolates from chickens, turkeys, pigeons, and cockatoos were also used as antigens. AVS-I had a high binding affinity for all La Sota and B1 strains, including vaccines. The antibody bound with a lower titer to the Australian Queensland V4 and Ulster strains, but it did not bind to the F strain, a lentogenic strain from England. AVS-I was HI-negative against the other PMV reference strains. AVS-I may be valuable for identifying field isolates antigenically similar to La Sota and B1 and rapidly differentiate those vaccine strains from more virulent viruses. 相似文献
20.
The humoral responses of Mycoplasma meleagridis-free and -infected turkeys were compared following primary and secondary antigenic stimulation with inactivated Salmonella pullorum or dinitrophenyl-bovine gamma globulin. The mycoplasma-infected groups had significantly lower antibody responses to both antigens. The suppression was more evident in the secondary response. The results suggest that the immune response in turkeys infected with M. meleagridis is similar to that of bursectomized chickens: the ability of chickens to synthesize immunoglobulin G is affected more readily than the ability to synthesize IgM. 相似文献